Tadashi Nakanishi, Kanako Takahashi, Kazumi Ozaki, Hideaki Nakae and Takashi Matsuo : Proceedings of the International Conference on Dentin/Pulp Complex 2001, --- An Immunohistologic Study on the Localization of Selected Bacteria and Chemokines in Human Deep-Carious Teeth ---, Quintessence Publishing CO.,Ltd., Tokyo, Jun. 2002.
Yoshitaka Hosokawa, Ikuko Hosokawa, Kazumi Ozaki and Takashi Matsuo : Nobiletin Inhibits Inflammatory Reaction in Interleukin-1β-Stimulated Human Periodontal Ligament Cells., Pharmaceutics, Vol.13, No.5, 667, 2021.
(要約)
The immune response in periodontal lesions is involved in the progression of periodontal disease. Therefore, it is important to find a bioactive substance that has anti-inflammatory effects in periodontal lesions. This study aimed to examine if nobiletin, which is found in the peel of citrus fruits, could inhibit inflammatory responses in interleukin (IL)-1β-stimulated human periodontal ligament cells (HPDLCs). The release of cytokines (IL-6, IL-8, CXCL10, CCL20, and CCL2) and matrix metalloproteinases (MMP-1 and MMP-3) was assessed by ELISA. The expression of cell adhesion molecules (ICAM-1and VCAM-1) and the activation of signal transduction pathways (nuclear factor (NF)-κB, mitogen-activated protein kinases (MAPKs) and protein kinase B (Akt)) in HPDLCs were detected by Western blot analysis. Our experiments revealed that nobiletin decreased the expression of inflammatory cytokines, cell adhesion molecules, and MMPs in IL-1β-stimulated HPDLCs. Moreover, we revealed that nobiletin treatment could suppress the activation of the NF-κB, MAPKs, and Akt pathways. These findings indicate that nobiletin could inhibit inflammatory reactions in IL-1β-stimulated HPDLCs by inhibiting multiple signal transduction pathways, including NF-κB, MAPKs, and Akt.
Yoshitaka Hosokawa, Ikuko Hosokawa, Kazumi Ozaki and Takashi Matsuo : The Polymethoxy Flavonoid Sudachitin Inhibits Interleukin-1 β-Induced Inflammatory Mediator Production in Human Periodontal Ligament Cells, BioMed Research International, Vol.2021, No.Article ID 8826586, 2021.
Ikuko Hosokawa, Yoshitaka Hosokawa, Kazumi Ozaki and Takashi Matsuo : Carnosic acid inhibits inflammatory cytokines production in human periodontal ligament cells., Immunopharmacology and Immunotoxicology, Vol.42, No.4, 373-378, 2020.
(要約)
The results of this study suggest that CA has anti-inflammatory effects in human periodontal ligament cells by inhibiting JNK, NF-κB and STAT3 pathways.
Hitomi Kuramoto, Kouji Hirao, Hiromichi Yumoto, Yuki Hosokawa, Tadashi Nakanishi, Daisuke Takegawa, Ayako Washio, Chiaki Kitamura and Takashi Matsuo : Caffeic acid phenethyl ester (CAPE) induces VEGF expression and production in rat odontoblastic cells, BioMed Research International, Vol.Article ID 5390720, 2019.
(要約)
Caffeic acid phenethyl ester (CAPE), the main component of propolis, has various biological activities including anti-inflammatory effect and wound healing promotion. Odontoblasts located in the outermost layer of dental pulp play crucial roles such as production of growth factors and formation of hard tissue termed reparative dentin in host defense against dental caries. In this study, we investigated the effects of CAPE on the upregulation of vascular endothelial growth factor (VEGF) and calcification activities of odontoblasts, leading to development of novel therapy for dental pulp inflammation caused by dental caries. CAPE significantly induced mRNA expression and production of VEGF in rat clonal odontoblast-like KN-3 cells cultured in normal medium or osteogenic induction medium. CAPE treatment enhanced nuclear factor-kappa B (NF-B) transcription factor activation, and furthermore, the specific inhibitor of NF-B significantly reduced VEGF production. The expression of VEGF receptor- (VEGFR-) 2, not VEGFR-1, was up regulated in KN-3 cells treated with CAPE. In addition, VEGF significantly increased mineralization activity in KN-3 cells. These findings suggest that CAPE might be useful as a novel biological material for the dental pulp conservative therapy.
Yoshitaka Hosokawa, Ikuko Hosokawa, Kazumi Ozaki and Takashi Matsuo : Sudachitin Inhibits Matrix Metalloproteinase-1 and -3 Production in Tumor Necrosis Factor-α-Stimulated Human Periodontal Ligament Cells., Inflammation, Vol.42, No.4, 1456-1462, 2019.
(要約)
Sudachitin, a polymethoxylated flavonoid found in the skin of Citrus sudachi, is a biologically active substance. The aim of this study was to examine whether sudachitin could be used to inhibit the expression of matrix metalloproteinase (MMP)-1 and MMP-3, which are involved in the destruction of periodontal tissues in periodontal lesions, in tumor necrosis factor (TNF)-α-stimulated human periodontal ligament cells (HPDLC). Sudachitin suppressed TNF-α-induced MMP-1 and MMP-3 production in HPDLC. On the other hand, it enhanced tissue inhibitor of metalloproteinase (TIMP)-1 expression. The level of Akt phosphorylation in the TNF-α-stimulated HPDLC was decreased by sudachitin treatment. Moreover, an Akt inhibitor reduced MMP-1 and MMP-3 production and increased TIMP-1 production. These findings indicate that sudachitin reduces MMP-1 and MMP-3 production in TNF-α-stimulated HPDLC by inhibiting the Akt pathway.
Ikuko Hosokawa, Yoshitaka Hosokawa, Kazumi Ozaki and Takashi Matsuo : Carnosic Acid Inhibits CXCR3 Ligands Production in IL-27-Stimulated Human Oral Epithelial Cells., Inflammation, Vol.42, No.4, 1311-1316, 2019.
(要約)
Carnosic acid, which is a bioactive compound isolated from rosemary, has various pharmacological effects. However, the anti-inflammatory effect of carnosic acid on periodontitis is still unknown. The aim of this study was to investigate the effect of carnosic acid on CXC chemokine receptor 3 (CXCR3) ligands, which are involved in Th1 cells migration and accumulation, production in interleukin (IL)-27-stimulated human oral epithelial cells (TR146 cells). Carnosic acid decreased CXC chemokine ligand (CXCL)9, CXCL10, and CXCL11 production in IL-27-stimulated TR146 cells in a dose-dependent fashion. Moreover, we disclosed that carnosic acid could suppress signal transducer and activator of transcription (STAT)1, STAT3, and protein kinase B (Akt) phosphorylation in IL-27-stimulated TR146 cells. Furthermore, STAT1, STAT3, and Akt inhibitors could suppress CXCR3 ligands production in IL-27-treated TR146 cells. In summary, carnosic acid could reduce CXCR3 ligands production in human oral epithelial cell by inhibiting STAT1, STAT3, and Akt activation.
Yoshitaka Hosokawa, Ikuko Hosokawa, Kazumi Ozaki and Takashi Matsuo : Honokiol and Magnolol Inhibit CXCL10 and CXCL11 Production in IL-27-Stimulated Human Oral Epithelial Cells., Inflammation, Vol.41, No.6, 2110-2115, 2018.
(要約)
Honokiol and magnolol, which are lignans isolated from Magnolia quinquepeta, have some pharmacological effects. However, the anti-inflammatory effects of honokiol and magnolol on periodontal disease are still uncertain. The aim of this study was to examine the effect of honokiol and magnolol on CXC chemokine receptor 3 (CXCR3) ligands, which are related with Th1 cell migration, production in interleukin (IL)-27-stimulated human oral epithelial cells (TR146 cells). Honokiol and magnolol inhibited CXC chemokine ligand (CXCL)10 and CXCL11 production in IL-27-stimulated TR146 cells in a dose-dependent manner. Moreover, we revealed that honokiol and magnolol could suppress signal transducer and activator of transcription (STAT)3 and protein kinase B (Akt) phosphorylation in IL-27-stimulated TR146 cells though STAT1 phosphorylation was not suppressed by honokiol and magnolol treatment. Furthermore, STAT3 and Akt inhibitors could suppress CXCR3 ligand production in TR146 cells. In summary, honokiol and magnolol could reduce CXCR3 ligand production in oral epithelial cell by inhibiting STAT3 and Akt activation.
Yoshihito Naitou, Hiromichi Yumoto, K Hs Kumar, Takashi Matsuo, Katsuhiko Hirota, Yoichiro Miyake, Kan Nagao, Yoritoki Tomotake, R Jimbo and Tetsuo Ichikawa : Antifungal and Mechanical Properties of Tissue Conditioner Containing Plant-Derived Component, --- An In Vitro Study ---, Journal of Prosthodontics, Vol.27, No.7, 665-669, 2018.
(要約)
To evaluate the antifungal activity and mechanical properties of a novel antifungal tissue conditioner containing Juncus powder. Juncus powder was mixed with GC tissue conditioner at concentrations of 2.5%, 5.0%, and 10.0% by mass. The cylindrical specimens of Juncus-mixed tissue conditioner (dimensions: 10 mm in diameter and 2 and 6 mm in height for antimicrobial and mechanical tests, respectively) were prepared. The specimens placed on the bottom of the 24-well tissue culture plate were cultured with Candida albicans CAD1 for 2 and 4 days. The proliferation of the C. albicans in the wells was determined by measuring the optical density of fungal culture, and the surface of the specimens were also observed by scanning electron microscopy (SEM). To assess the mechanical properties of the specimens, the fluidity and hardness of Juncus-mixed tissue conditioner were measured using the methods certified according to ISO 10139-1. Juncus-mixed tissue conditioner significantly exhibited growth inhibitory effect in a Juncus concentration-dependent manner after both 2- and 4- day cultures. SEM observation showed that the amount of C. albicans on Juncus-mixed specimens drastically decreased, and biofilm formation was markedly inhibited. Moreover, both mechanical properties were found to be within the ranges regulated and specified by ISO. These findings demonstrated that the tissue conditioner including Juncus powder has a significant growth inhibitory effect against C. albicans, and it is suggested that the application of Juncus-mixed tissue conditioner may prevent denture stomatitis and oral candidiasis in denture wearers.
Yoshitaka Hosokawa, Ikuko Hosokawa, Kazumi Ozaki and Takashi Matsuo : Transforming growth factor-β1 increases C-C chemokine ligand 11 production in interleukin 4-stimulated human periodontal ligament cells., Cell Biology International, Vol.42, No.10, 1395-1400, 2018.
(要約)
Transforming growth factor (TGF)-β1 is a multifunctional cytokine, which can control certain functions of various kinds of cells. However, it is unclear whether TGF-β1 affects T-cell migration in periodontal lesions. The aim of this study was to examine the effects of TGF-β1 on the production of C-C chemokine ligand (CCL)11, which is a T-helper 2-type chemokine, in human periodontal ligament cells (HPDLC). Interleukin (IL)-4 induced CCL11 production, but TGF-β1 did not, in HPDLC. However, TGF-β1 enhanced CCL11 production in IL-4-stimulated HPDLC. Western blot analysis showed that the signal transducer and activator of transcription 6 (STAT6) pathway was highly activated in HPDLC that had been stimulated with both IL-4 and TGF-β1. Mitogen-activated protein kinase activation did not differ between the HPDLC treated with a combination of IL-4 and TGF-β1 and those treated with IL-4 or TGF-β1 alone. Moreover, a STAT6 inhibitor significantly inhibited CCL11 production in HPDLC that had been stimulated with IL-4 and TGF-β1. The current study clearly demonstrated that TGF-β1 enhanced IL-4-induced CCL11 production in HPDLC. The STAT6 pathway is important for CCL11 production in IL-4- and TGF-β1-treated HPDLC.
Susilowati Heni, Keiji Murakami, Hiromichi Yumoto, Takashi Amoh, Kouji Hirao, Katsuhiko Hirota, Takashi Matsuo and Yoichiro Miyake : Royal jelly inhibits Pseudomonas aeruginosa adherence and reduces excessive inflammatory responses in human epithelial cells, BioMed Research International, Vol.2017, 3191752, 2017.
(要約)
is a Gram-negative bacterium and causes respiratory infection especially in elderly patients. Royal jelly has been used worldwide as a traditional remedy and as a nutrient; however, the effect against is unclear. The aim of this study was to analyze antibacterial, antiadherent, and anti-inflammatory effects of royal jelly against . Wild-type strain PAO1 and clinical isolates of were used for antibacterial assay and antiadherent assay to abiotic surface and epithelial cells, which are pharynx (Detroit 562) and lung (NCI-H292) epithelial cells. In anti-inflammatory assay, epithelial cells were pretreated with royal jelly before bacterial exposure to investigate its inhibitory effect on interleukin (IL-8) and macrophage inflammatory protein-3/CCL20 overproduction. Although royal jelly did not have antibacterial activity at concentration of 50% w/v, antiadherent activity was confirmed on the abiotic surface and epithelial cells under concentration of 25%. Pretreatment with royal jelly significantly inhibited overproduction of IL-8 and CCL20 from both cells. These results demonstrated that royal jelly inhibits adherence and protects epithelial cells from excessive inflammatory responses against infection. Our findings suggested that royal jelly may be a useful supplement as complementary and alternative medicine for preventing respiratory infection caused by .
Sekita Yasuko, Keiji Murakami, Hiromichi Yumoto, Kouji Hirao, Takashi Amoh, Natsumi Fujiwara, Katsuhiko Hirota, Hideki Fujii, Takashi Matsuo, Yoichiro Miyake and Yoshiki Kashiwada : Antibiofilm and Anti-Inflammatory Activities of+ Houttuynia cordata Decoction for Oral Care, Evidence-Based Complementary and Alternative Medicine : eCAM, Vol.2017, 2850947, 2017.
(要約)
Dental biofilms that form in the oral cavity play a critical role in the pathogenesis of several infectious oral diseases, including dental caries, periodontal disease, and oral candidiasis. (HC, Saururaceae) is a widely used traditional medicine, for both internal and external application. A decoction of dried HC leaves (dHC) has long been consumed as a health-promoting herbal tea in Japan. We have recently reported that a water solution of HC poultice ethanol extract (wHCP) exerts antimicrobial and antibiofilm effects against several important oral pathogens. It also exhibits anti-inflammatory effects on human keratinocytes. In our current study, we examined the effects of dHC on infectious oral pathogens and inflammation. Our results demonstrated that dHC exerts moderate antimicrobial effects against methicillin-resistant (MRSA) and other oral microorganisms. dHC also exhibited antibiofilm effects against MRSA, (involved in dental plaque formation), and and inhibitory effects on interleukin-8, CCL20, IP-10, and GRO productions by human oral keratinocytes stimulated by lipopolysaccharide (a cause of periodontal disease), without cytotoxic effects. This suggests that dHC exhibits multiple activities in microorganisms and host cells. dHC can be easily prepared and may be effective in preventing infectious oral diseases.
Yoshitaka Hosokawa, Ikuko Hosokawa, Satoru Shindo, Kazumi Ozaki and Takashi Matsuo : IL-29 Enhances CXCL10 Production in TNF--stimulated Human Oral Epithelial Cells., Immunological Investigations, Vol.46, No.6, 615-624, 2017.
(要約)
Interleukin-29 (IL-29) is a cytokine belonging to the Type III interferon family. It was recently detected in the gingival crevicular fluid of periodontitis patients. However, the role of IL-29 in the pathogenesis of periodontal disease remains unknown. The aim of this study was to examine the effects of IL-29 on C-X-C motif chemokine ligand 10 (CXCL10) production in human oral epithelial cells. We measured CXCL10 production in TR146 cells, which is a human oral epithelial cell line, using an enzyme-linked immunosorbent assay. We used a Western blot analysis to detect IL-29 receptor expression and the phosphorylation levels of signal transduction molecules, including p38 mitogen-activated protein kinases (MAPK), signal transducer and activator of transcription 3 (STAT3), and nuclear factor (NF)- B p65, in the TR146 cells. The TR146 cells expressed the IL-29 receptor. IL-29 induced CXCL10 production in the TR146 cells. IL-29 significantly enhanced CXCL10 production in tumor necrosis factor (TNF)--stimulated TR146 cells. The p38 MAPK, STAT3, and NF-B pathways were found to be related to the IL-29-induced enhancement of CXCL10 production in TNF--stimulated TR146 cells. IL-29 promotes T helper 1-cell accumulation in periodontal lesions by inducing CXCL10 production in oral epithelial cells.
Halitosis is caused by volatile sulphur compounds including methyl mercaptan (CH3 SH) in the oral cavity and is a serious problem that limits interpersonal social communication. The aim of study was to evaluate the effects of reuterin-related compounds (RRCs) on halitosis-related periodontopathic bacteria in vitro. RRC-01, RRC-02 and RRC-03 (32 and 64 g ml(-1) ) in culture media containing Fusobacterium nucleatum JCM8523 and Porphyromonas gingivalis ATCC33277 were used. The effects of RRCs on CH3 SH production and detectable odour by F. nucleatum and P. gingivalis were examined by CH3 SH production assay and organoleptic test, respectively. The number of bacterial cells was also measured using an ATP assay. In P. gingivalis treated with RRCs, the expression of mgl gene, which is responsible for CH3 SH production, was examined by qRT-PCR. CH3 SH production and the score of detectable odour from F. nucleatum and P. gingivalis culture media containing RRCs were significantly lower than that without RRCs (P < 0.05). The expression of mgl gene in P. gingivalis was significantly downregulated by RRC-01 (P < 0.01), but not by RRC-02 or RRC-03. RRCs are potent oral care products for preventing halitosis via reducing CH3 SH production.
Yoshitaka Hosokawa, Ikuko Hosokawa, Satoru Shindo, Kazumi Ozaki and Takashi Matsuo : Gomisin N Decreases Inflammatory Cytokine Production in Human Periodontal Ligament Cells., Inflammation, Vol.40, No.2, 360-365, 2017.
(要約)
Gomisin N, which is a lignan isolated from Schisandra chinensis, has some pharmacological effects. However, the anti-inflammatory effects of gomisin N on periodontal disease are uncertain. The aim of this study was to examine the effect of gomisin N on inflammatory mediator production in tumor necrosis factor (TNF)--stimulated human periodontal ligament cells (HPDLC). Gomisin N inhibited interleukin (IL)-6, IL-8, CC chemokine ligand (CCL) 2, and CCL20 production in TNF--stimulated HPDLC in a dose-dependent manner. Moreover, we revealed that gomisin N could suppress extracellular signal-regulated kinase (ERK) and c-Jun N terminal kinase (JNK) phosphorylation in TNF--stimulated HPDLC though protein kinase B (Akt) phosphorylation was not suppressed by gomisin N treatment. In summary, gomisin N might exert anti-inflammatory effects by attenuating cytokine production in periodontal ligament cells via inhibiting the TNF--stimulated ERK and JNK pathways activation.
Keiji Murakami, Hiromichi Yumoto, Ayu Murakami, Takashi Amoh, Darija Viducic, Katsuhiko Hirota, Atsushi Tabata, Hideaki Nagamune, Hiroki Kourai, Takashi Matsuo and Yoichiro Miyake : Evaluation of the effectiveness of the potent bis-quaternary ammonium compound, 4,4'-( , -hexametylenedithio) bis (1-octylpyridinium bromide) (4DTBP-6,8) on Pseudomonas aeruginosa., Journal of Applied Microbiology, Vol.122, No.4, 893-899, 2017.
(要約)
Quaternary ammonium compounds (QACs), including benzalkonium chloride (BAC) and cetylpyridinium chloride (CPC) are cationic surfactants and have been used widely as general disinfectants in the medical field due to their strong antibacterial effects and low cytotoxicity to human cells. 4,4'-( , -hexametylenedithio) bis (1-octylpyridinium bromide) (4DTBP-6,8) is one of the potent bis-QACs synthesized to improve the antimicrobial activities of mono-QACs such as BAC. This study aimed to assess the effectiveness of 4DTBP-6,8 against Pseudomonas aeruginosa, a prevalent hospital pathogen. The minimum inhibitory concentrations of 4DTBP-6,8, CPC and BAC against P. aeruginosa were measured. 4DTBP-6,8 exhibited strong antibacterial activity. We assessed the bactericidal effects of QACs against P. aeruginosa under certain conditions and their cytotoxicities in human epithelial cells using lactate dehydrogenase (LDH) release. 4DTBP-6,8 exerted excellent bactericidal effects against high concentrations of bacteria, biofilm cells and even in the presence of contaminated proteins. Cellular LDH was not released by the treatment with 4DTBP-6,8. 4DTBP-6,8 exhibited the strongest bactericidal activity against P. aeruginosa among the three QACs tested without any cytotoxicity. The potent bis-QAC, 4DTBP-6,8 has the potential to be an effective disinfectant in preventing hospital infections caused by P. aeruginosa.
Katsuhiko Hirota, Hiromichi Yumoto, B Sapaar, Takashi Matsuo, Tetsuo Ichikawa and Yoichiro Miyake : Pathogenic factors in Candida biofilm-related infectious diseases., Journal of Applied Microbiology, Vol.122, No.2, 321-330, 2016.
(要約)
Candida albicans is a commonly found member of the human microflora and is a major human opportunistic fungal pathogen. A perturbation of the microbiome can lead to infectious diseases caused by various micro-organisms, including C. albicans. Moreover, the interactions between C. albicans and bacteria are considered to play critical roles in human health. The major biological feature of C. albicans, which impacts human health, resides in its ability to form biofilms. In particular, the extracellular matrix (ECM) of Candida biofilm plays a multifaceted role and therefore may be considered as a highly attractive target to combat biofilm-related infectious diseases. In addition, extracellular DNA (eDNA) also plays a crucial role in Candida biofilm formation and its structural integrity and induces the morphological transition from yeast to the hyphal growth form during C. albicans biofilm development. This review focuses on pathogenic factors such as eDNA in Candida biofilm formation and its ECM production and provides meaningful information for future studies to develop a novel strategy to battle infectious diseases elicited by Candida-formed biofilm.
Yoshitaka Hosokawa, Ikuko Hosokawa, Satoru Shindo, Yoshihiro Ohta, Kazumi Ozaki and Takashi Matsuo : Alkannin inhibits CCL3 and CCL5 production in human periodontal ligament cells., Cell Biology International, Vol.40, No.12, 1380-1385, 2016.
(要約)
Alkannin, which is found in Alkanna tinctoria, a member of the borage family, is used as a food coloring. Alkannin has recently been reported to have certain biological functions, such as anti-microbial and anti-oxidant effects. It is known that CC chemokine receptor (CCR) 5-positive leukocytes contribute to alveolar bone resorption in periodontal lesions. The aim of this study was to examine whether alkannin inhibits the production of CC chemokine ligand (CCL) 3 and CCL5, which are CCR5 ligands, in human periodontal ligament cells (HPDLC). Interleukin (IL)-1 induced CCL3 and CCL5 production in HPDLC. Alkannin inhibited IL-1-mediated CCL3 and CCL5 production in HPDLC in a dose-dependent manner. Moreover, we revealed that alkannin suppressed inhibitor of kappa B- degradation in IL-1-stimulated HPDLC. In addition, a nuclear factor (NF)-B inhibitor significantly inhibited CCL3 and CCL5 production in IL-1-stimulated HPDLC. These results demonstrate that alkannin inhibits CCR5 ligand production in IL-1-stimulated HPDLC by attenuating the NF-B signaling pathway.
Yuki Hosokawa, Kouji Hirao, Hiromichi Yumoto, Ayako Washio, Tadashi Nakanishi, Daisuke Takegawa, Chiaki Kitamura and Takashi Matsuo : Functional roles of NOD1 in odontoblasts on dental pulp innate immunity, BioMed Research International, Vol.2016, 9325436, 2016.
(要約)
Caries-related pathogens are first recognized by odontoblasts and induce inflammatory events that develop to pulpitis. Generally, initial sensing of microbial pathogens is mediated by pattern recognition receptors, such as Toll-like receptor and nucleotide-binding oligomerization domain (NOD); however, little is known about NODs in odontoblasts. In this study, the levels of NODs expressed in rat odontoblastic cell line, KN-3, were assessed by flow cytometry and the levels of chemokines in NOD-specific ligand-stimulated KN-3 cells were analyzed by real-time PCR and ELISA. The signal transduction pathway activated with NOD-specific ligand was assessed by blocking assay with specific inhibitors and reporter assay. In KN-3 cells, the expression level of NOD1 was stronger than that of NOD2 and the production of chemokines, such as CINC-1, CINC-2, CCL20, and MCP-1, was upregulated by stimulation with NOD1-specific ligand, but not with NOD2-specific ligand. CINC-2 and CCL20 production by stimulation with NOD1-specific ligand was reduced by p38 MAPK and AP-1 signaling inhibitors. Furthermore, the reporter assay demonstrated AP-1 activation in NOD1-specific ligand-stimulated KN-3 cells. These findings indicated that NOD1 expressed in odontoblasts functions to upregulate the chemokines expression via p38-AP-1 signaling pathway and suggested that NOD1 may play important roles in the initiation and progression of pulpitis.
Ikuko Hosokawa, Yoshitaka Hosokawa, Satoru Shindo, Kazumi Ozaki and Takashi Matsuo : Melatonin Inhibits CXCL10 and MMP-1 Production in IL-1-Stimulated Human Periodontal Ligament Cells., Inflammation, Vol.39, No.4, 1520-1526, 2016.
(要約)
Melatonin is a hormone that is mainly secreted by the pineal gland and exhibits a wide spectrum of activities, including antioxidant functions. Melatonin has been detected in gingival crevicular fluid. However, the role of melatonin in periodontal tissue is still uncertain. The aim of this study was to examine the effects of melatonin on inflammatory mediator expression in human periodontal ligament cells (HPDLC). Interleukin (IL)-1 induced CXC chemokine ligand (CXCL)10, matrix metalloproteinase (MMP)-1, and tissue inhibitors of metalloproteinase (TIMP)-1 production in HPDLC. Melatonin decreased CXCL10 and MMP-1 production and increased TIMP-1 production in IL-1-stimulated HPDLC. Western blot analysis showed that melatonin inhibited p38 mitogen-activated protein kinase (MAPK) and c-jun N-terminal kinase (JNK) phosphorylation, and IkB- degradation and phosphorylation in IL-1-stimulated HPDLC. These results suggest that melatonin might inhibit Th1 cell migration by reducing CXCL10 production. Moreover, melatonin might inhibit soft tissue destruction by decreasing MMP-1 production in periodontal lesions.
Yasuko Sekita, Keiji Murakami, Hiromichi Yumoto, Takashi Amoh, Natsumi Fujiwara, Shohei Ogata, Takashi Matsuo, Yoichiro Miyake and Yoshiki Kashiwada : Preventive Effects of Houttuynia cordata Extract for Oral Infectious Diseases., BioMed Research International, Vol.2016, 2581876, 2016.
(要約)
Houttuynia cordata (HC) (Saururaceae) has been used internally and externally as a traditional medicine and as an herbal tea for healthcare in Japan. Our recent survey showed that HC poultice (HCP) prepared from smothering fresh leaves of HC had been frequently used for the treatment of purulent skin diseases with high effectiveness. Our experimental study also demonstrated that ethanol extract of HCP (eHCP) has antibacterial, antibiofilm, and anti-inflammatory effects against S. aureus which caused purulent skin diseases. In this study, we focused on novel effects of HCP against oral infectious diseases, such as periodontal disease and dental caries. We determined the antimicrobial and antibiofilm effects of water solution of HCP ethanol extract (wHCP) against important oral pathogens and investigated its cytotoxicity and anti-inflammatory effects on human oral epithelial cells. wHCP had moderate antimicrobial effects against some oral microorganisms and profound antibiofilm effects against Fusobacterium nucleatum, Streptococcus mutans, and Candida albicans. In addition, wHCP had no cytotoxic effects and could inhibit interleukin-8 and CCL20 productions by Porphyromonas gingivalis lipopolysaccharide-stimulated human oral keratinocytes. Our findings suggested that wHCP may be clinically useful for preventing oral infectious diseases as a mouthwash for oral care.
Satoru Shindo, Yoshitaka Hosokawa, Ikuko Hosokawa, Kazumi Ozaki and Takashi Matsuo : Shikonin Inhibits Inflammatory Cytokine Production in Human Periodontal Ligament Cells., Inflammation, Vol.39, No.3, 1124-1129, 2016.
(要約)
Shikonin, which is derived from Lithospermum erythrorhizon, a herb used in traditional medicine, has long been considered to be a useful treatment for various diseases in traditional oriental medicine. Shikonin has recently been reported to have several pharmacological properties, e.g., it has anti-microbial, anti-tumor, and anti-inflammatory effects. The aim of this study was to examine whether shikonin is able to influence the production of interleukin (IL)-6, IL-8, and/or chemokine C-C motif ligand (CCL)20, which contribute to the pathogenesis of periodontal disease, in human periodontal ligament cells (HPDLC). The production levels of IL-6, IL-8, and CCL20 in HPDLC were determined using an ELISA. Western blot analysis was used to detect nuclear factor kappa B (NF-B) pathway activation in HPDLC. Shikonin prevented IL-1- or tumor necrosis factor (TNF)--mediated IL-6, IL-8, and CCL20 production in HPDLC. Moreover, we found that shikonin suppressed the phosphorylation and degradation of inhibitor of kappa B-alpha (IB-) in IL-1- or TNF--stimulated HPDLC. These findings suggest that shikonin could have direct beneficial effects against periodontal disease by reducing IL-6, IL-8, and CCL20 production in periodontal lesions.
Yasuko Sekita, Keiji Murakami, Hiromichi Yumoto, Hiroyuki Mizuguchi, Takashi Amoh, Satoshi Ogino, Takashi Matsuo, Yoichiro Miyake, Hiroyuki Fukui and Yoshiki Kashiwada : Anti-bacterial and anti-inflammatory effects of ethanol extract from Houttuynia cordata poultice., Bioscience, Biotechnology, and Biochemistry, 2016.
(要約)
Houttuynia cordata (HC) has been commonly used as many traditional remedies in local areas of Japan. Although many pharmacological activities of HC have been reported, the mechanism underlying the effect of HC remains unknown. We conducted the interview survey in Japan to verify how HC was actually used. The interview survey revealed that HC poultice (HCP) prepared from smothering fresh leaves of HC was most frequently used for the treatment of purulent skin diseases including furuncle and carbuncle with high effectiveness. Ethanol extract of HCP (eHCP) showed anti-bacterial effects against methicillin-resistant Staphylococcus aureus (MRSA), and showed an anti-biofilm activity against MRSA. eHCP showed dose-dependent inhibition of S. aureus lipoteichoic acid (LTA)-induced interleukin-8 and CCL20 production in human keratinocyte without any cytotoxicity. These results suggest that HCP is effective for skin abscess and its underlying mechanism might be the complicated multiple activities for both bacteria and host cells.
Tsuyoshi Miyagawa, Tsuyoshi Fujita, Hiromichi Yumoto, Tetsuya Yoshimoto, Mikihito Kajiya, Kazuhisa Ouhara, Shinji Matsuda, Hideki Shiba, Takashi Matsuo and Hidemi Kurihara : Azithromycin recovers reductions in barrier function in human gingival epithelial cells stimulated with tumor necrosis factor-α, Archives of Oral Biology, Vol.62, 64-69, 2016.
(要約)
The gingival epithelium plays an important role in protecting against the invasion of periodontal pathogens, and the permeability of gingival epithelial cells has been implicated in the initiation of periodontitis. Azithromycin (AZM) has been used in the treatment of chronic inflammatory airway diseases because it regulates cell-cell contact in airway epithelial cells. Therefore, AZM may also regulate barrier function in gingival epithelial cells. In the present study, we examined the effects of AZM on the permeability of human gingival epithelial cells (HGEC) under inflammatory conditions in vitro. HGEC were stimulated by tumor necrosis factor-α (TNF-α) in the presence of AZM or p38 MAP kinase and ERK inhibitors. Permeability was assessed based on transepithelial electrical resistance (TER). The expression of E-cadherin, phosphorylated p38 MAP kinase, and ERK was analyzed by Western blotting. TNF-α decreased TER in HGEC, and AZM and the p38 MAP kinase and ERK inhibitors recovered this decrease. AZM inhibited the phosphorylation of ERK and p38 MAP kinase in TNF-α-stimulated HGEC. Furthermore, AZM recovered the decrease in E-cadherin expression in HGEC stimulated with TNF-α. These results suggested that AZM regulated gingival epithelial permeability through p38 MAP kinase and ERK signaling, and may contribute to suppress the inflammation in gingival tissue.
Yoshitaka Hosokawa, Ikuko Hosokawa, Satoru Shindo, Kazumi Ozaki and Takashi Matsuo : IL-4 Modulates CCL11 and CCL20 Productions from IL-1-Stimulated Human Periodontal Ligament Cells., Cellular Physiology and Biochemistry, Vol.38, No.1, 153-159, 2016.
(要約)
These results mean that IL-4 enhanced Th2 cells migration in periodontal lesion to induce CCL11 production from HPDLCs. On the other hand, IL-4 inhibits Th17 cells accumulation in periodontally diseased tissues to inhibit CCL20 production. Therefore, IL-4 is positively related with the pathogenesis of periodontal disease to control chemokine productions in periodontal lesions.
Yoshitaka Hosokawa, Ikuko Hosokawa, Satoru Shindo, Kazumi Ozaki and Takashi Matsuo : Calcitriol Suppressed Inflammatory Reactions in IL-1-Stimulated Human Periodontal Ligament Cells., Inflammation, Vol.38, No.6, 2252-2258, 2015.
(要約)
Vitamin D has important roles on control of calcium and phosphate levels in the body. However, the role of vitamin D on the pathogenesis of periodontal disease is still uncertain. Therefore, we examined the effect of the hormonal form of vitamin D, calcitriol, on inflammatory responses of human periodontal ligament cells (HPDLC). We detected vitamin D receptor expression in non-stimulated HPDLC. Calcitriol inhibited interleukin (IL)-6, IL-8, CC chemokine ligand (CCL) 20, CXC chemokine ligand (CXCL) 10, and matrix metalloproteinase (MMP)-3 release from IL-1-stimulated HPDLC. Tissue inhibitor of metalloproteinase (TIMP)-1 production did not change by calcitriol. Moreover, we found c-jun N-terminal kinase (JNK) phosphorylation and IB- degradation in IL-1-stimulated HPDLC were inhibited by calcitriol, and JNK and nuclear factor (NF)-B inhibitors could decrease IL-6, IL-8, CCL20, CXCL10, and MMP-3 productions in IL-1-treated HPDLC. These findings suggest that vitamin D could modulate inflammatory response in periodontal tissues.
Tadashi Nakanishi, Kayo Mukai, Yoshitaka Hosokawa, Daisuke Takegawa and Takashi Matsuo : Catechins inhibit vascular endothelial growth factor production and cyclooxygenase-2 expression in human dental pulp cells, International Endodontic Journal, Vol.48, No.3, 277-282, 2015.
(要約)
To investigate the effect of catechins on vascular endothelial growth factor (VEGF) production and cyclooxygenase-2 (COX-2) expression in human dental pulp cells (HDPC) stimulated with bacteria-derived factors or pro-inflammatory cytokines. Morphologically fibroblastic cells established from explant cultures of healthy human dental pulp tissues were used as HDPC. HDPC pre-treated with catechins, epigallocatechin-3-gallate (EGCG) or epicatechin gallate (ECG), were exposed to lipopolysaccharide (LPS), peptidoglycan (PG), interlukin-1β (IL-1β) or tumour necrosis factor-α (TNF-α). VEGF production was examined by enzyme-linked immunosorbent assay, and COX-2 expression was assessed by immunoblot. EGCG and ECG significantly reduced LPS- or PG-mediated VEGF production in the HDPC in a dose-dependent manner. EGCG also prevented IL-1β-mediated VEGF production. Although TNF-α did not enhance VEGF production in the dental pulp cells, treatment of 20 μg mL(-1) of EGCG decreased the level of VEGF. In addition, the catechins attenuated COX-2 expression induced by LPS and IL-1β. The up-regulated VEGF and COX-2 expressions in the HDPC stimulated with these bacteria-derived factors or IL-1β were diminished by the treatment of EGCG and ECG. These findings suggest that the catechins may be beneficial as an anti-inflammatory tool of the treatment for pulpal inflammation.
Hiromichi Yumoto, Kouji Hirao, Toshihiko Tominaga, Naoki Bando, Kanako Takahashi and Takashi Matsuo : Electromagnetic wave irradiation promotes osteoblastic cell proliferation and up-regulates growth factors via activation of the ERK1/2 and p38 MAPK pathways, Cellular Physiology and Biochemistry, Vol.35, No.2, 601-615, 2015.
(要約)
Periodontitis with bone resorption is caused by inflammatory reactions to bacterial infection. We recently reported that electromagnetic wave irradiation (EMWI) has bactericidal effects. However, the effects of EMWI on periodontal tissues remain unclear. This study was aimed to investigate the effects of EMWI on osteoblasts. Osteoblastic cells MC3T3-E1 were treated with EMWI (500-1,000 kHz, 5 times, 1 sec/time). Cell growth and cytotoxicity were determined by cell proliferation assays and measurement of lactate dehydrogenase release, respectively. Gene expression and protein production of growth factors were analyzed using real-time PCR and ELISA, respectively. EMWI-activated cellular signal transduction pathways were investigated by immunoblotting and blocking assay with specific inhibitors. Osteoblasts proliferation was significantly enhanced 3 days after EMWI and no cytotoxicity was observed. EMWI up-regulated various growth factors, such as vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF). EMWI induced ERK1/2, p38 MAPK and SAPK/JNK phosphorylation within 5 min, and the production of PDGF-ΑΑ and VEGF was partially reduced by MAPK-specific inhibitor. These findings demonstrated that EMWI increases osteoblastic cell activity and the expression of growth factors via ERK1/2 and p38 MAPK pathways and suggested that EMWI may be beneficial to bone tissue repair such as periodontitis.
It has been shown that oral hygiene affects the onset of perioperative complications. The usefulness of perioperative oral function management aiming at the outbreak decrease in treatment complications and an early discharge was recognized. As a result, perioperative oral function management fee was founded at revision of medical service fees in Fiscal year 2012. In this clinical study, we evaluated the implementation of perioperative oral function management in Tokushima University Hospital. We examined 781 patients, including 563 patients for surgery and 218 patients for chemotherapy and radiotherapy. The mean age of patients was 58.8 ± 12.4 years old. The implementation rate of perioperative oral function management was 9.7% in the patients of surgery, and 17.4% in those of chemotherapy and radiotherapy. The highly required medical department was neurosurgery in the patients of surgery, and hematology in those of chemotherapy and radiotherapy. The mean number of tooth present was 21.3 ± 7.1 in the patients of surgery, and 19.8 ± 7.2 in those chemotherapy and radiotherapy. The rate of dental treatment was required in 40.5% of total patients who received surgery, and in 51.4% of patients who received chemotherapy and radiotherapy.The rate of patients who received denture treatment attained to 11.9% of the whole patients receiving surgery, and 13.3% of patients receiving chemotherapy and radiotherapy. It was revealed that there were many patients required potential demands in perioperative oral function management, and that there were many patients who need dental or denture treatment. We would like to develop perioperative oral function management by the interprofessional collaboration in health and social care.7月発行だが，6月号である．
Satoru Shindo, Yoshitaka Hosokawa, Ikuko Hosokawa, Kazumi Ozaki and Takashi Matsuo : Genipin inhibits MMP-1 and MMP-3 release from TNF--stimulated human periodontal ligament cells., Biochimie, Vol.107PB, 391-395, 2014.
(要約)
Genipin, the aglycon of geniposide found in gardenia fruit has long been considered for treatment of inflammatory diseases in traditional oriental medicine. Genipin has recently been reported to have some pharmacological functions, such as antimicrobial, antitumor, and anti-inflammatory effects. The aim of this study was to examine whether genipin could modify matrix metalloproteinase (MMP)-1 and MMP-3, which are related to the destruction of periodontal tissues in periodontal lesion, expression in tumor necrosis factor (TNF)--stimulated human periodontal ligament cells (HPDLCs). Genipin prevented TNF--mediated MMP-1 and MMP-3 productions in HPDLCs. Moreover, genipin could suppress not only extracellular signal-regulated kinase (ERK) and Jun-N-terminal kinase (JNK) phosphorylations but also AMP-activated protein kinase (AMPK) phosphorylation in TNF--stimulated HPDLCs. Inhibitors of ERK and AMPK could inhibit both MMP-1 and MMP-3 productions. Moreover, we revealed the ERK inhibitor suppressed AMPK phosphorylation in TNF--stimulated HPDLCs. These data provide a new mechanism through which genipin could be used for the treatment of periodontal disease to prevent MMPs expression in periodontal lesion.
Yoshitaka Hosokawa, Ikuko Hosokawa, Satoru Shindo, Kazumi Ozaki and Takashi Matsuo : IL-22 Enhances CCL20 Production in IL-1-Stimulated Human Gingival Fibroblasts., Inflammation, Vol.37, No.6, 2062-2066, 2014.
(要約)
CC chemokine ligand 20 (CCL20) is involved in the recruitment of Th17 cells and thus in the exacerbation of periodontal disease, but the effect of simultaneous interleukin (IL)-22 and IL-1 stimulation on CCL20 production in human gingival fibroblasts (HGFs) is uncertain. In this study, we investigated the mechanisms of IL-1- and/or IL-22-induced CCL20 production in HGFs. A single stimulation of IL-22 could not induce CCL20 production. On the other hand, IL-22 could increase CCL20 production from IL-1-stimulated HGFs in a dose-dependent manner. C-Jun N terminal kinase (JNK) and inhibitor of nuclear factor B (IB)- phosphorylation were increased in IL-1- and IL-22-stimulated HGFs. An inhibitor of nuclear factor (NF)-B decreased IL-1- and IL-22-induced CCL20 production, though an inhibitor of JNK did not modulate CCL20 production. These data suggest that IL-1 in cooperation with IL-22 could increase Th17 cell accumulation in periodontally diseased tissues to enhance CCL20 production in HGFs.
Daisuke Takegawa, Tadashi Nakanishi, Kouji Hirao, Hiromichi Yumoto, Kanako Takahashi and Takashi Matsuo : Modulatory roles of interferon-γ through indoleamine 2, 3-dioxygenase induction in innate immune response of dental pulp cells., Journal of Endodontics, Vol.40, No.9, 1382-1387, 2014.
(要約)
Marked infiltration of inflammatory cells such as activated T cells producing interferon-γ (IFN-γ) is observed in severe pulpitis. However, the roles of IFN-γ in the innate immune response of dental pulp have not been reported. Indoleamine 2, 3-dioxygenase (IDO) is a regulator of immune responses, and the IDO expression is induced by IFN-γ in many cells whose expression in dental pulp is unknown. The purpose of this study was to determine the role of IFN-γ in the immune response through microbial pattern recognition receptors (PRRs) such as Toll-like receptors or nucleotide-binding oligomerization domain-like receptors on the production of proinflammatory cytokines such as CXCL10 and interleukin (IL)-6 and the expression of IDO in cultured human dental pulp cells (HDPCs). HDPCs were established from explant cultures of healthy pulp tissues. CXCL10 and IL-6 production was determined using enzyme-linked immunosorbent assay. Confirmation of IDO localization in dental pulp tissues was examined using immunohistochemistry. IDO expression in HDPCs was analyzed by immunoblot. IFN-γ significantly up-regulated CXCL10 and IL-6 production in the HDPCs stimulated with ligands for PRRs in a concentration-dependent manner. The expression of IDO was detected in inflamed pulp tissue. In addition, IFN-γ in combination with the PRR ligands enhanced IDO expression in HDPCs compared with IFN-γ alone. Moreover, CXCL10 production in IFN-γ-stimulated HDPCs was inhibited by an IDO inhibitor. This study showed the synergistic effects by IFN-γ on cytokine production and IDO expression in HDPCs, suggesting that IFN-γ may modulate the innate immune response of dental pulp.
Nur Asikin, Katsuhiko Hirota, Hiromichi Yumoto, Kouji Hirao, Kanako Takahashi, Keiji Murakami, Takashi Matsuo and Yoichiro Miyake : Effect of extracellular DNA on pyocyanin production from Pseudomonas aeruginosa, Bacterial Adherence & Biofilm, Vol.27, 45-47, 2014.
41.
Yoshitaka Hosokawa, Satoru Shindo, Ikuko Hosokawa, Kazumi Ozaki and Takashi Matsuo : IL-6 trans-signaling enhances CCL20 production from IL-1-stimulated human periodontal ligament cells., Inflammation, Vol.37, No.2, 381-386, 2014.
(要約)
CC chemokine ligand 20 (CCL20) plays a central role in the recruitment of CCR6-expressing cells, including Th17 cells which are related to bone resorption in periodontal lesions and thus in the development of periodontal disease. IL-6 is an important cytokine that is associated with the pathogenesis of periodontitis. However, the effect of IL-6 on CCL20 production is uncertain. The aim of this study was to examine whether IL-6 could modify CCL20 expression in human periodontal ligament cells (HPDLCs). HPDLCs expressed gp130 but did not express IL-6R on the surface of HPDLCs. So, IL-6 trans-signaling is important to recognize IL-6 by HPDLCs. IL-6/sIL-6R stimulation enhanced CCL20 production in IL-1-stimulated HPDLCs. IL-6 produced from IL-1-stimulated HPDLCs with sIL-6R could increase CCL20 production in HPDLCs with sIL-6R. Signal transducer and activator of transcription (STAT)3 activation was related to CCL20 production in IL-1 and IL-6/sIL-6R-stimulated HPDLCs. Our data suggests that HPDLCs, in response to IL-6, sIL-6R, and IL-1, may shift chemokine production to that favoring CCR6-expressing cells recruitment in periodontal lesions.
B Sapaar, A Nur, Katsuhiko Hirota, Hiromichi Yumoto, Keiji Murakami, T Amoh, Takashi Matsuo, Tetsuo Ichikawa and Yoichiro Miyake : Effects of extracellular DNA from Candida albicans and pneumonia-related pathogens on Candida biofilm formation and hyphal transformation., Journal of Applied Microbiology, Vol.116, No.6, 1531-1542, 2014.
(要約)
The aim of this study was to investigate the effects of genomic DNA purified from Candida albicans and pneumonia-related pathogens, Pseudomonas aeruginosa and Staphylococcus aureus, on in vitro biofilm formation and morphological change of 3 Candida species (C. albicans, C. glabrata, and C. tropicalis). Biofilm formation was evaluated by the crystal violet assay and colony-forming unit counts. Morphological characteristics of biofilms were evaluated by scanning electron microscopy and fluorescence microscopy. Addition of DNA at a low concentration (<1·0 μg ml(-1)) significantly increased biofilm mass of all three Candida species. In contrast, the addition of DNA at a high concentration (10 μg ml(-1)) decreased the biofilm mass. Interestingly, the formation of hyphae in a dense network of yeast cells was observed in C. albicans biofilms exposed to a low concentration of DNA (<1·0 μg ml(-1)). These findings demonstrated that extracellular DNA (eDNA) plays a crucial role in Candida biofilm formation and suggested that eDNA may induce the morphological transition from yeast to hyphal growth form during C. albicans biofilm development. A novel therapy targeting eDNA may be applicable for Candida infection to decrease biofilm formation and hyphal formation.
Satoru Shindo, Yoshitaka Hosokawa, Ikuko Hosokawa, Kazumi Ozaki and Takashi Matsuo : Genipin inhibits IL-1-induced CCL20 and IL-6 production from human periodontal ligament cells., Cellular Physiology and Biochemistry, Vol.33, No.2, 357-364, 2014.
(要約)
These data provide a novel mechanism through which genipin could be used to provide direct benefits in periodontal disease to inhibit IL-6 and CCL20 productions in periodontal lesions.
Wang Yiwei, 湯本 浩通, Liu Dali, 松尾 敬志, Shu Rong : The comparison of the ability of polysaccharide from different Porphyromonas gingivalis in inducing THP-1 cells to produce cytokines, Chinese Journal of Stomatology, Vol.49, No.2, 78-83, 2014年.
(要約)
To compare the ability of the polysaccharide from various Porphyromonas gingivalis (Pg) type and clinical strains in inducing THP-1 cells to produce cytokines interleukin(IL)-1β, IL-8, and tumor necrosis factor(TNF)-α, in order to analyze the immunogenicity of Pg polysaccharide components and the virulence-associated factors of this periodontal pathogen. The bacterial polysaccharide was extracted from high virulent Pg strains, W83, SJD2, SJD12 and low virulent Pg, ATCC33277, SJD4, SJD5, and SJD11 by phenol-water extraction. The extracted polysaccharide was used to stimulate the THP-1 cells with different simulation periods and doses. The level of the cytokines, including IL-1β,IL-8 and TNF-α in the cell culture suspension was measured by enzyme-linked immunosorbent assay(ELISA). The polysaccharide extraction of Pg strains was composed of lipopolysaccharide(LPS) and capsular polysaccharide. The secretion of IL-1β, IL-8 and TNF-α, produced by the THP-1 cells showed in a time- and dose-dependent manner in the medium containing 10% fetal bovine serum. The level of these cytokines of the high virulent strains was higher than that of the low virulent strains in medium containing 1% fetal bovine serum.Four hours after stimulation with polysaccharide extracted from high virulent strains, the levels of IL-1β,IL-8, and TNF-α in the cell suspension were (1 639 ± 497), (1 648 ± 513) and (140 ± 48) µg/L, respectively, whereas for low virulent strains, the levels of IL-1β, IL-8, and TNF-α were (773 ± 382), (892 ± 400) and (67 ± 33) µg/L, respectively. Polysaccharide extracted from Pg could induced the THP-1 cells to secrete the cytokines of IL-1β, IL-8 and TNF-α. The level of the cytokines produced by the THP-1 cells associates with the bacterial virulent properties.
Yoshitaka Hosokawa, Ikuko Hosokawa, Satoru Shindo, Kazumi Ozaki and Takashi Matsuo : (-)-Epigallocatechin-3-Gallate Inhibits CC Chemokine Ligand 11 Production in Human Gingival Fibroblasts., Cellular Physiology and Biochemistry, Vol.31, No.6, 960-967, 2013.
(要約)
Background: CC chemokine ligand 11 (CCL11) is related to Th2 cells migration via CC chemokine receptor 3 (CCR3). Th2 cells are involved in the etiology of periodontal disease. However, how the infiltration of Th2 cells is controlled in periodontally diseased tissues is unknown. (-)-Epigallocatechin gallate (EGCG), the major catechin in green tea, has multiple beneficial effects, but the effects of EGCG on CCL11 production are uncertain. In this study, we investigated whether cytokines could induce CCL11 production in human gingival fibroblasts (HGFs). Moreover, we examined the effects of EGCG on CCL11 production in HGFs. Methods and Results: ELISA analysis disclosed that interleukin (IL)-4 synergistically enhanced CCL11 production in IL-1 or tumor necrosis factor (TNF)--stimulated HGFs. EGCG prevented IL-1/ IL-4 or TNF-/IL-4-mediated CCL11 production in a concentration dependent manner. CCL11 production in HGFs was positively regulated by p38 mitogen-activated protein kinase (MAPK), extracellular signal-regulated kinase (ERK), and c-Jun N terminal kinase (JNK). Western blot analysis revealed that EGCG treatment prevented IL-1/IL-4 or TNF-/IL-4-induced ERK and JNK activation in HGFs. Conclusions: These data provide that CCL11 production in HGFs could be associated with Th2 cells infiltration in periodontal lesions. Moreover, EGCG is useful for periodontitis treatment to inhibit CCL11 production.
Yoshitaka Hosokawa, Ikuko Hosokawa, Satoru Shindo, Kazumi Ozaki and Takashi Matsuo : TLR3 agonist enhances CC chemokine ligand 20 production in IL-1-stimulated human gingival fibroblasts., Cellular Immunology, Vol.283, No.1-2, 8-11, 2013.
(要約)
Viruses are related to the etiology of periodontitis. However, the role of viruses on Th17 cells infiltration in periodontitis lesions is unknown. Therefore, we examined the effects of TLR3 ligand on CCL20, which is related to Th17 cells migration, production in human gingival fibroblasts (HGFs). Polyinosinic-polycytidylic acid (Poly I:C), which is a TLR3 agonist, stimulation could moderately induce CCL20 production in HGFs. Poly I:C synergistically enhanced CCL20 expression from IL-1-stimulated HGFs. Inhibitors of p38 MAPK, extracellular signal-regulated kinase (ERK), c-Jun N terminal kinase (JNK), and NF-B significantly inhibited CCL20 production in Poly I:C/IL-1-stimulated HGFs. Western blot analysis disclosed phosphorylation of p38 MAPK, JNK, and IB- were enhanced in Poly I:C/IL-1-treated HGFs. These data suggested that virus infection is related to Th17 cells migration in periodontitis lesion to induce CCL20 production in HGFs via TLR3. Therefore, our results indicated that virus might be important pathogen in periodontal disease.
Nur Asikin, Katsuhiko Hirota, Hiromichi Yumoto, Kouji Hirao, Kanako Takahashi, Keiji Murakami, Takashi Matsuo and Yoichiro Miyake : Role of Extracellular DNA and DNA-binding protein in biofilm formation of Streptococcus intermeius, Bacterial Adherence & Biofilm, Vol.26, 25-29, 2013.
48.
Nur Asikin, Katsuhiko Hirota, Hiromichi Yumoto, Kouji Hirao, D Liu, Kanako Takahashi, Keiji Murakami, Takashi Matsuo, R Shu and Yoichiro Miyake : Effects of extracellular DNA and DNA-binding protein on the development of a Streptococcus intermedius biofilm., Journal of Applied Microbiology, Vol.115, No.1, 260-270, 2013.
(要約)
The aim of this study was to clarify the effects of homologous and heterologous extracellular DNAs (eDNAs) and histone-like DNA-binding protein (HLP) on Streptococcus intermedius biofilm development and rigidity. Formed biofilm mass was measured with 0·1% crystal violet staining method and observed with a scanning electron microscope. The localizations of eDNA and extracellular HLP (eHLP) in formed biofilm were detected by staining with 7-hydoxyl-9H-(1,3-dichloro-9,9-dimethylacridin-2-one) and anti-HLP antibody without fixation, respectively. DNase I treatment (200 U ml(-1) ) markedly decreased biofilm formation and cell density in biofilms. Colocalization of eHLP and eDNA in biofilm was confirmed. The addition of eDNA (up to 1 μg ml(-1) ) purified from Strep. intermedius, other Gram-positive bacteria, Gram-negative bacteria, or human KB cells into the Strep. intermedius culture increased the biofilm mass of all tested strains of Strep. intermedius, wild-type, HLP-downregulated strain and control strains. In contrast, the addition of eDNA (>1 μg ml(-1) ) decreased the biofilm mass of all Strep. intermedius strains. These findings demonstrated that eDNA and eHLP play crucial roles in biofilm development and its rigidity. eDNA- and HLP-targeting strategies may be applicable to novel treatments for bacterial biofilm-related infectious diseases.
(キーワード)
Biofilms / Cell Line, Tumor / DNA (DNA) / DNA-Binding Proteins / Deoxyribonuclease I / Humans / Streptococcus intermedius
Toshiyuki Suge, Ishikawa Kunio and Takashi Matsuo : Changes in the crystallinity of hydroxyapatite powder and structure of enamel treated with several concentrations of ammonium hexafluorosilicate, American Journal of Dentistry, Vol.25, No.5, 299-302, 2012.
(要約)
To evaluate the effects of changing concentrations of ammonium hexafluorosilicate [SiF: (NH4)2SiF6] solution on the crystallinity of hydroxyapatite powder (HAP) and structure of human enamel in order to overcome the tooth discoloration caused by diamine silver fluoride [AgF: (NH3)2AgF] application. HAP was treated with several concentrations of SiF solution (from 10 to 19,400 ppm) for 5 minutes. The crystallinity of the HAP before and after SiF treatment was then measured by powder X-ray diffraction (XRD) analysis. The angular width (beta) of the 002 diffraction peak was measured at 1/2 the height of the maximum intensity. Also, enamel specimens were prepared from a human extracted tooth. Several concentrations of SiF solution were applied to polished or phosphoric acid etched enamel specimens. The enamel surface was then observed using scanning electron microscopy (SEM). XRD peaks became sharper after SiF treatment indicating that the crystallinity of apatite powder was increased. The 1/beta value was increased from 2.8 +/- 0.1 to 4.3 +/- 0.1 after treatment with 1,000 ppm SiF solution. The amount of CaF2 formed in HAP was gradually increased with increasing concentrations of SiF solution. TheXRD pattern was consistent with CaF2 in case of over 9,000 ppm SiF solution. SEM photographs demonstrated that exposed enamel rods with acid etching were filled with CaF2-like precipitate after SiF treatment regardless of the concentration of SiF solution.
Yoshitaka Hosokawa, Ikuko Hosokawa, Satoru Shindo, Kazumi Ozaki, Hideaki Nakae and Takashi Matsuo : Tumor necrosis factor-like weak inducer of apoptosis increases CC chemokine ligand 20 production in interleukin 1-stimulated human gingival fibroblasts., Human Immunology, Vol.73, No.5, 470-473, 2012.
(要約)
CC chemokine ligand 20 (CCL20) is related to T-helper (Th)-17 cell migration, and Th17 cells play important roles in exacerbation in periodontal disease. However, the effect of tumor necrosis factor-like weak inducer of apoptosis (TWEAK) on CCL20 production is unknown. In this study, we examined the mechanisms of TWEAK in combination with interleukin (IL)-1-induced CCL20 production in human gingival fibroblasts (HGFs). TWEAK alone did not induce CCL20 production in HGFs. However, TWEAK enhanced CCL20 expression from IL-1-stimulated HGFs in a dose-dependent manner. Inhibitors of p38 mitogen-activated protein kinase, extracellular signal-regulated kinase (ERK), protein kinase B (Akt), and nuclear factor B (NF-B) significantly inhibited CCL20 production in TWEAK and IL-1-stimulated HGFs. Western blot analysis revealed that phosphorylations of ERK, Akt, and inhibitor of NF-B were enhanced in TWEAK and IL-1-treated HGFs. These data suggest that TWEAK is positively related to Th17 cell migration in periodontally diseased tissues to enhance CCL20 production in IL-1-stimulated HGFs.
Tsuyoshi Fujita, Hiromichi Yumoto, Hideki Shiba, Kazuhisa Ouhara, Tsuyoshi Miyagawa, Takayoshi Nagahara, Shinji Matsuda, Hiroyuki Kawaguchi, Takashi Matsuo, Shinya Murakami and Hidemi Kurihara : Irsogladine maleate regulates epithelial barrier function in TNF--stimulated human gingival epithelial cells, Journal of Periodontal Research, Vol.47, No.1, 55-61, 2012.
(要約)
As epithelial cells function as a mechanical barrier, the permeability of the gingival epithelial cell layer indicates a defensive capability against invasion by periodontal pathogens. We have reported the expression of claudin-1 and E-cadherin, key regulators of permeability, in the gingival junctional epithelium. Irsogladine maleate (IM) is a medication for gastric ulcers and also regulates Aggregatibacter actinomycetemcomitans-stimuated chemokine secretion and E-cadherin expression in gingival epithelium. In this study, we have further investigated the effects of IM on the barrier functions of gingival epithelial cells under inflammatory conditions. We examined the permeability, and the expression of claudin-1 and E-cadherin, in human gingival epithelial cells (HGECs) stimulated with tumor necrosis factor (TNF)-α, with or without IM. TNF-α increased the permeability of HGECs, and IM abolished the increase. TNF-α reduced the expression of E-cadherin in HGECs, and IM reversed the reduction. In addition, immunofluorescence staining showed that TNF-α disrupted claudin-1 expression in HGECs, and IM reversed this effect. The results suggest that IM reverses the TNF-α-induced disruption of the gingival epithelial barrier by regulating E-cadherin and claudin-1.
Shibata Shingo, Toshiyuki Suge, Tomoko Kimura, Ishikawa Kunio and Takashi Matsuo : Antibacterial activity of ammonium hexafluorosilicate solution with antimicrobial agents for the prevention of dentin caries, American Journal of Dentistry, Vol.25, No.1, 31-34, 2012.
(要約)
This study evaluated the antibacterial activity of the SiF solution with the addition of antibacterial agents on a Streptococcus mutans biofilm. Various antibacterial SiF solutions were prepared by adding chlorhexidine, cetylpyridinium chloride, isopropyl methylphenol, or epigallocatechin gallate. Hydroxyapatite pellets treated with several SiF solutions were immersed in BHI inoculated with S. mutans standardized suspension. The number of S. mutans cells adhered to each pellet was evaluated. SiF with the addition of CPC was the most effective for reducing the adherence of bacteria and inhibiting the formation ofbiofilm, showing the same level as AgF, In contrast, the addition of other antibacterial agents to SiF reduced the original antibacterial activity of SiF solution.
Yoshitaka Hosokawa, Ikuko Hosokawa, Kazumi Ozaki, Hideaki Nakae and Takashi Matsuo : Interleukin (IL)-17A synergistically enhances CC chemokine ligand 20 production in IL-1-stimulated human gingival fibroblasts., Human Immunology, Vol.73, No.1, 26-30, 2012.
(要約)
CC chemokine ligand 20 (CCL20) plays a pivotal role in the recruitment of T-helper (Th)-17 cells and thus in the development of periodontal disease, but the effect of simultaneous interleukin (IL)-17A and IL-1 stimulation on CCL20 production in human gingival fibroblasts (HGFs) are not known. In this study, we investigated the mechanisms of IL-1- and IL-17A-induced CCL20 production in HGFs. IL-17A synergistically enhanced CCL20 production from IL-1-stimulated HGFs in a concentration-dependent manner. Extracellular signal-regulated kinase (ERK) and inhibitor of nuclear factor (NF)-B- phosphorylation were increased in IL-1- and IL-17A-stimulated HGFs. Inhibitors of or ERK and NF-B decreased IL-1- and IL-17A-induced CCL20 production. IL-1 stimulation elevated IL-17 receptor C expression on HGFs. These data suggest that IL-1 is actively related to Th17 cell migration into peripheral tissues to induce production of the Th17 chemokine, CCL20. Therefore, IL-1 might be a therapeutic target for Th17-related diseases, such as periodontal disease and arthritis.
Tadashi Nakanishi, Daisuke Takegawa, Kouji Hirao, Kanako Takahashi, Hiromichi Yumoto and Takashi Matsuo : Roles of dental pulp fibroblasts in the recognition of bacterium-related factors and subsequent development of pulpitis, Japanese Dental Science Review, Vol.47, No.2, 161-166, 2011.
Shinya Horiuchi, Shingo Kuroda, Masahiro Hiasa, Toshiyuki Suge, Seitaro Saku, Kenichi Hamada, Takashi Matsuo, Kenzo Asaoka and Eiji Tanaka : Reinforcement of bond strength of self-etching orthodontic adhesive., The Angle Orthodontist, Vol.82, No.1, 30-35, 2011.
(要約)
Abstract Objective: To determine the reinforcement of bond strength of a self-etching system by applying a pretreatment agent. Materials and Methods: Sixty extracted human premolars were used in this study. The enamel surfaces were treated with four pretreatment agents?phosphoric acid, polyacrylic acid, citric acid, and ammonium hexafluorosilicate (SiF)?and were examined under a scanning electron microscope. Afterward, orthodontic brackets were bonded with a self-etching adhesive system (n ?=? 10 for each agent), and shear bond strength was measured through a debonding process. The adhesive remnant index (ARI) was also assessed. Results: Enamel surfaces treated with polyacrylic acid seemed almost the same as intact enamel. Treatment with SiF induced slight shallow depressions compared with the intact enamel. On the other hand, enamel surfaces treated with citric acid and phosphoric acid showed severe etching patterns. All pretreatments increased the bond strength, but SiF-treated specimens revealed the greatest strength (12.201 ? 1.048?MPa), followed by polyacrylic acid (12.030 ? 2.103?MPa). The control group with no pretreatment showed the least strength (9.078 ? 1.678?MPa). All pretreatments increased ARI score compared with the control group. Conclusions: Surface conditioning before bracket adhesion could reinforce the bond strength of the self-etching adhesive system, resulting in a more reliable bonding system.
Yoshitaka Hosokawa, Ikuko Hosokawa, Satoru Shindo, Kazumi Ozaki, Tadashi Nakanishi, Hideaki Nakae and Takashi Matsuo : Black tea polyphenol inhibits CXCL10 production in oncostatin M-stimulated human gingival fibroblasts., International Immunopharmacology, Vol.11, No.6, 670-674, 2011.
(要約)
CXC chemokine ligand 10 (CXCL10) plays an important role in the infiltration of Th1 cells and thus in the exacerbation of periodontal disease. Theaflavin-3,3'-digallate (TFDG), polyphenol in black tea, has some beneficial effects but the effect of TFDG on CXCL10 production from human gingival fibroblasts (HGFs) is uncertain. In this study, we investigated the mechanisms by which TFDG may inhibit oncostatin M (OSM)-induced CXCL10 production in human gingival fibroblasts. TFDG prevented OSM-mediated CXCL10 production by HGFs in a dose dependent manner. TFDG significantly inhibited OSM-induced phosphorylation of c-Jun N terminal kinase (JNK), protein kinase B (Akt) (Ser473) that are related to CXCL10 production from OSM-stimulated HGFs. In addition, TFDG suppressed OSM receptor (OSMR) β expression on HGFs. These data provide a novel mechanism where the black tea flavonoid, theaflavin, could provide direct benefits in periodontal disease.
Katsuhiko Hirota, Hiromichi Yumoto, K Miyamoto, N Yamamoto, Keiji Murakami, Yumi Hoshimo, Takashi Matsuo and Yoichiro Miyake : MPC-Polymer Reduces Adherence and Biofilm Formation by Oral Bacteria., Journal of Dental Research, Vol.90, No.7, 900-905, 2011.
(要約)
Oral biofilms such as dental plaque cause dental caries and periodontitis, as well as aspiration pneumonia and infectious endocarditis by translocation. Hence, the suppression of oral biofilm formation is an issue of considerable importance. Mechanical removal, disinfectants, inhibition of polysaccharide formation, and artificial sugar have been used for the reduction of oral biofilm. From the viewpoint of the inhibition of bacterial adherence, we investigated whether aqueous biocompatible 2-methacryloyloxyethyl phosphorylcholine (MPC)-polymer can reduce streptococcal colonization and biofilm formation. We examined the effects of MPC-polymer on streptococcal adherence to saliva-coated hydroxyapatite and oral epithelial cells, and the adherence of Fusobacterium nucleatum to streptococcal biofilm. MPC-polymer application markedly inhibited both the adherence and biofilm formation of Streptococcus mutans on saliva-coated hydroxyapatite and streptococcal adherence to oral epithelial cells, and reduced the adherence of F. nucleatum to streptococcal biofilms. A small-scale clinical trial revealed that mouthrinsing with MPC-polymer inhibited the increase of oral bacterial numbers, especially of S. mutans. These findings suggest that MPC-polymer is a potent inhibitor of bacterial adherence and biofilm development, and may be useful to prevent dental-plaque-related diseases. (UMIN Clinical Trial Registry UMIN000003471). Abbreviations: QOL, quality of life; MPC, 2-methacryloyloxyethyl phosphorylcholine; BHI, brain-heart infusion broth; PBS, phosphate-buffered saline; SEM, scanning electron microscopy; CFU, colony-forming unit.
J Li, Katsuhiko Hirota, Hiromichi Yumoto, Takashi Matsuo, Yoichiro Miyake and Tetsuo Ichikawa : Enhanced germicidal effects of pulsed UV-LED irradiation on biofilms., Journal of Applied Microbiology, Vol.109, No.6, 2183-2190, 2010.
(要約)
The germicidal effects of UVA-LED irradiation (365 nm, 0·28 mW cm(-2) , in pulsed or continuous mode) on Candida albicans or Escherichia coli biofilms were evaluated by determining colony-forming units. The morphological change of microbial cells in biofilms was observed using scanning electron microscopy. After 5-min irradiation, over 90% of viable micro-organisms in biofilms had been killed, and pulsed irradiation (1-1000 Hz) had significantly greater germicidal ability than continuous irradiation. Pulsed irradiation (100 Hz, 60 min) almost completely killed micro-organisms in biofilm (>99·9%), and 20-min irradiation greatly damaged both microbial species. Interestingly, few hyphae were found in irradiated Candida biofilms. Moreover, mannitol treatment, a scavenger of hydroxyl radicals (OH(•) ), significantly protected viable micro-organisms in biofilms from UVA-LED irradiation.
Yoshitaka Hosokawa, Ikuko Hosokawa, Kazumi Ozaki, Hideaki Nakae and Takashi Matsuo : Oncostatin M synergistically induces CXCL10 and ICAM-1 expression in IL-1beta-stimulated-human gingival fibroblasts., Journal of Cellular Biochemistry, Vol.111, No.1, 40-48, 2010.
(要約)
Periodontitis is a chronic bacterial infection of tooth-supporting structures. T-helper type 1 (Th1) cells are related to the exacerbation of periodontal disease. Human gingival fibroblasts (HGFs), the major cell type in periodontal connective tissues, are involved in immunological response in periodontal tissues. However, it is uncertain whether HGFs are related to Th1 response. Chemokine (C-X-C motif) ligand 10 (CXCL10) is a cytokine, that is related to Th1 cells migration. Intercellular adhesion molecule (ICAM)-1 is involved in Th1 cells retention and activation in inflamed tissue. The aim of this study is to examine the effect of oncostatin M (OSM) on CXCL10 and ICAM-1 expression in HGFs. OSM stimulation induced CXCL10 and ICAM-1 expression in HGFs. Moreover, the synergistic effects of CXCL10 release and ICAM-1 expression in HGFs were observed with combined stimulation of interleukin (IL)-1beta and OSM. OSM increased type 1 IL-1 receptor (IL-1R1) expression, and IL-1beta enhanced OSMRbeta expression on HGFs. IL-1beta + OSM stimulation enhanced the phosphorylation of inhibitor of nuclear factor kappaB (IkappaB)-alpha, signal transducer and activator of transcription (STAT)3, c-Jun N terminal kinase (JNK), and protein kinase B (Akt) compared to OSM or IL-1beta stimulation. CXCL10 production from OSM + IL-1beta stimulated HGFs was suppressed by nuclear factor (NF)-kappaB, STAT3, JNK, and phosphoinositide-3-kinase (PI3K) inhibitors. On the other hand, only NF-kappaB and STAT3 inhibitors suppressed ICAM-1 expression enhanced by OSM + IL-1beta treatment. These effects of OSM and IL-1beta may promote Th1 cells infiltration and retention in periodontally diseased tissues and be related to exacerbation of periodontal disease. J. Cell. Biochem. 111: 40-48, 2010. (c) 2010 Wiley-Liss, Inc.
Yoshitaka Hosokawa, Ikuko Hosokawa, Kazumi Ozaki, Tadashi Nakanishi, Hideaki Nakae and Takashi Matsuo : Tea polyphenols inhibit IL-6 production in tumor necrosis factor superfamily 14-stimulated human gingival fibroblasts., Molecular Nutrition & Food Research, Vol.54 Suppl 2, S151-8, 2010.
(要約)
IL-6 is well recognized to be a potent bone resorptive agent and thus in the development of periodontal disease. Epigallocatechin gallate (EGCG) and epicatechin gallate (ECG), the major catechins in green tea, and theaflavin-3,3'-digallate (TFDG), polyphenol in black tea, have multiple beneficial effects, but the effects of catechins and theaflavins on IL-6 production in human gingival fibroblasts (HGFs) are not known. In this study, we investigated the mechanisms by which EGCG, ECG, and TFDG inhibit tumor necrosis factor superfamily 14 (TNFSF14)-induced IL-6 production in HGFs. We detected TNFSF14 mRNA expression in human diseased periodontal tissues. TNFSF14 increased IL-6 production in HGFs in a concentration-dependent manner. EGCG, ECG, and TFDG prevented TNFSF14-mediated IL-6 production in HGFs. EGCG, ECG, and TFDG prevented TNFSF14-induced extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and nuclear factor-kappaB activation in HGFs. Inhibitors of ERK, JNK, and nuclear factor-kappaB decreased TNFSF14-induced IL-6 production. In addition, EGCG, ECG, and TFDG attenuated TNFSF14 receptor expression on HGFs. These data provide a novel mechanism through which the green tea and black tea polyphenols could be used to provide direct benefits in periodontal disease.
Yoshitaka Hosokawa, Ikuko Hosokawa, Kazumi Ozaki, Tadashi Nakanishi, Hideaki Nakae and Takashi Matsuo : Catechins inhibit CXCL10 production from oncostatin M-stimulated human gingival fibroblasts, The Journal of Nutritional Biochemistry, Vol.21, No.7, 659-664, 2010.
(要約)
CXC chemokine ligand 10 (CXCL10) plays a pivotal role in the recruitment of Th1 cells and, thus, in the development of periodontal disease. Epigallocatechin gallate (EGCG) and epicatechin gallate (ECG), the major catechins derived from green tea, have multiple beneficial effects, but the effects of catechins on CXCL10 production from human gingival fibroblasts (HGFs) is not known. In this study, we investigated the mechanisms by which EGCG and ECG inhibit oncostatin M (OSM)-induced CXCL10 production in HGFs. HGFs constitutively expressed glycoprotein 130 and OSM receptor beta (OSMR beta), which are OSM receptors. OSM increased CXCL10 production in a concentration-dependent manner. EGCG and ECG prevented OSM-mediated CXCL10 production by HGFs. Inhibitors of p38 mitogen-activated protein kinase, c-Jun N-terminal kinase (JNK), phosphatidylinositol-3-OH kinase and signal transducer and activator of transcription (STAT)3 decreased OSM-induced CXCL10 production. EGCG significantly prevented OSM-induced phosphorylation of JNK, Akt (Ser473) and STAT3 (Tyr705 and Ser727). ECG prevented phosphorylation of JNK and Akt (Ser473). In addition, EGCG and ECG attenuated OSMR beta expression on HGFs. These data provide a novel mechanism through which the green tea flavonoids, catechins, can provide direct benefits in periodontal disease.
Tadashi Nakanishi, Kayo Mukai, Hiromichi Yumoto, Kouji Hirao, Yoshitaka Hosokawa and Takashi Matsuo : Anti-inflammatory effect of catechin on cultured dental pulp cells affected by bacteria-derived factors, European Journal of Oral Sciences, Vol.118, No.2, 145-150, 2010.
(要約)
Catechins (bioactive polyphenols in green tea) are known to exhibit potent anti-inflammatory properties. However, the anti-inflammatory effects of catechins on inflamed dental pulp tissue are not known. In this study, we investigated the effect of epigallocatechin-3-gallate (EGCG) and epicatechin gallate (ECG), the major components of green tea catechins, on the expression of pro-inflammatory cytokines and adhesion molecules in human dental pulp cells stimulated with bacteria-derived factors such as lipopolysaccharide (LPS) and peptidoglycan (PG). The expression of interleukin (IL)-6 and of IL-8 was examined using the reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assays. The expression of intercellular adhesion molecule-1 (ICAM-1) and of vascular cell adhesion molecule-1 (VCAM-1) on dental pulp cells was analyzed using flow cytometry. The presence of EGCG and ECG significantly reduced, in a concentration-dependent manner, the expression of IL-6 and IL-8 in dental pulp cells exposed to LPS or PG. Increased expression of ICAM-1 and VCAM-1 on the dental pulp cells in response to bacterial components was also decreased by treatment with EGCG and ECG. These findings suggest that green tea catechins may prevent the exacerbation of pulpitis.
Ikuko Hosokawa, Yoshitaka Hosokawa, Kazumi Ozaki, Hiromichi Yumoto, Hideaki Nakae and Takashi Matsuo : Proinflammatroy effects of muramyldipeptide on human gingival fibroblasts, Journal of Periodontal Research, Vol.45, No.2, 193-199, 2010.
(要約)
Because human gingival fibroblasts (HGFs) are the predominant cells in periodontal tissues, we hypothesized that HGFs are contributed to receptors for components of bacteria. In this study, we focused on expression and function of nucleotide binding oligomerization domain 2 (NOD2) in HGFs, which is a mammalian cytosolic pathogen recognition molecule. Expression of NOD2 in HGFs was examined by reverse transcriptase-polymerase chain reaction (RT-PCR) and flow cytometry. Production of interleukin (IL)-6, IL-8, cc chemokine ligand2, cxc chemokine ligand10 (CXCL10) and CXCL11 from HGFs was examined by enzyme-linked immunosorbent assay (ELISA). We used RT-PCR and immunohistochemistry to detect the NOD2 expression in human gingival tissues. We found clear NOD2 expression in HGFs. Upon stimulation with NOD2 agonist, muramyldipeptide (MDP), production of proinflammatory cytokines was enhanced. Moreover, MDP-induced production of proinflammatory cytokines was inhibited in a different manner by mitogen-activated protein kinase inhibitors and phosphatidylinositol 3-kinase inhibitor. Furthermore, MDP enhanced CXCL10 and CXCL11 productions by tumor necrosis factor-alpha (TNF-alpha)- or interferon-gamma (IFN-gamma)-stimulated HGFs, although MDP alone did not induce these chemokines. TNF-alpha and IFN-gamma increased NOD2 expression in HGFs. In addition, we detected NOD2 expression in mononuclear cells and HGFs in periodontally diseased tissues. These findings indicate that MDP which induces production of cytokines and chemokines from HGFs is related to the pathogenesis of periodontal disease.
Toshiyuki Suge, Akiko Kawasaki, Ishikawa Kunio, Takashi Matsuo and Ebisu Shigeyuki : Effects of ammonium hexafluorosilicate concentration on dentin tubule occlusion and composition of the precipitate, Dental Materials, Vol.26, No.1, 29-34, 2010.
(要約)
Ammonium hexafluorosilicate [SiF: (NH(4))(2)SiF(6)] was prepared in order to overcome the tooth discoloration caused by diamine silver fluoride [AgF: (NH(3))(2)AgF] application. We employed a single concentration of SiF solution in our previous study; therefore, it is still unclear how the concentration of SiF solution affects the occlusion of dentin tubules and composition of the precipitate. The aim of this study was to evaluate the effects of changing the concentration of SiF on its clinical use as a dentin hypersensitivity treatment. To simulate dentin tubules subject to dentin hypersensitivity, dentin disks were treated with EDTA for 2 min. Then, the disks were treated with several concentrations of SiF solution (from 100 to 19,400 ppm) for 3 min. The occlusion of dentin tubules was evaluated using scanning electron microscopy (SEM), and the composition of the precipitate formed in the tubules after SiF treatment was assessed using energy dispersive X-ray analysis (EDXA). SEM photographs demonstrated that dentin tubules after treatment with SiF were occluded homogeneously and fully regardless of the concentration of SiF solution. The Ca/P molar ratio of the precipitate formed in dentin tubules after SiF treatment was increased with the concentration of SiF solution. It was concluded that the capacity to occlude dentin tubules was the same regardless of the concentration of SiF solution. However, the composition of the precipitate formed in the tubules was dependent on the concentration of SiF solution.
Yoshitaka Hosokawa, Ikuko Hosokawa, Kazumi Ozaki, Hideaki Nakae and Takashi Matsuo : TNFSF14 coordinately enhances CXCL10 and CXCL11 production from IFN-gamma-stimulated human gingival fibroblasts, Molecular Immunology, Vol.47, No.4, 666-670, 2010.
(要約)
TNFSF14 is involved in the pathogenesis of some inflammatory diseases such as arthritis. CXCL10 and CXCL11 recruit Th1 cells, and the productions of these chemokines are related to the exacerbation of some inflammatory diseases including arthritis and periodontal disease. We examined in vitro effects of TNFSF14 on IFN-gamma-induced CXCL10 and CXCL11 production in human gingival fibroblasts (HGFs). HGFs constitutively expressed TNFSF14 receptors, LTbetaR and HVEM. TNFSF14 enhanced IFN-gamma-induced secretion of CXCL10 and CXCL11 from HGFs. IFN-gamma treatment increased HVEM expression on HGFs. TNFSF14 in combination with IFN-gamma resulted in increased activation of p38 MAPK, ERK and IkappaB-alpha compared with TNFSF14 or IFN-gamma alone. Moreover, inhibitors of p38 MAPK, ERK and NF-kappaB abolished the CXCL10 and CXCL11 productions from TNFSF14 with IFN-gamma-stimulated HGFs. These effects of TNFSF14 may promote the infiltration of Th1 cells into lesions with inflammatory diseases. TNFSF14 might act as a proinflammatory cytokine in some inflammatory diseases thus is a candidate therapeutic target.
Yoshitaka Hosokawa, Ikuko Hosokawa, Kazumi Ozaki, Tadashi Nakanishi, Hideaki Nakae and Takashi Matsuo : Catechins Inhibit CCL20 Production in IL-17A-stimulated Human Gingival Fibroblasts, Cellular Physiology and Biochemistry, Vol.24, No.5-6, 391-396, 2009.
(要約)
CC chemokine ligand 20 (CCL20) plays a pivotal role in the recruitment of Th17 cells and thus in the development of periodontal disease. Epigallocatechin gallate (EGCG) and epicatechin gallate (ECG), the major catechins in green tea, have multiple beneficial effects, but the effects of catechins on CCL20 production in human gingival fibroblasts (HGFs) are not known. In this study, we investigated the mechanisms by which EGCG and ECG inhibit interleukin (IL)-17A-induced CCL20 production in human gingival fibroblasts. IL-17A increased CCL20 production in HGFs in a concentration-dependent manner. EGCG and ECG prevented IL-17A-mediated CCL20 production in HGFs. Inhibitors of p38 mitogen-activated protein kinase (MAPK) or extracellular signal-regulated kinase (ERK) decreased IL-17A-induced CCL20 production. EGCG and ECG prevented IL-17A-induced phosphorylation of p38 MAPK and ERK in HGFs. In addition, EGCG and ECG attenuated IL-17 receptor expression on HGFs. These data provide a novel mechanism through which the green tea flavonoids catechins could be used to provide direct benefits in periodontal disease.
Kouji Hirao, Hiromichi Yumoto, Kanako Takahashi, Kayo Mukai, Tadashi Nakanishi and Takashi Matsuo : Roles of TLR2, TLR4, NOD2, and NOD1 in pulp fibroblasts., Journal of Dental Research, Vol.88, No.8, 762-767, 2009.
(要約)
Pulp fibroblasts express various pro-inflammatory mediators leading to marked infiltration of inflammatory cells in the progression of pulpitis. We hypothesized that pulp fibroblasts play roles in the recognition of invaded caries-related bacteria and the subsequent innate immune responses. We found clear expressions of TLR2, NOD1, and NOD2 and a faint expression of TLR4 in human dental pulp fibroblasts (HDPF) by RT-PCR and flow cytometry. We also observed that various pro-inflammatory mediators, including cytokines, chemokines, adhesion molecules, prostaglandin E(2) and its key enzyme COX-2, not iNOS or caspase-1, were markedly up-regulated by stimulation with these TLR and NOD agonists. More over, the NOD2 agonist acted synergistically with the TLR2, not the TLR4, agonist to stimulate the production of pro-inflammatory mediators in HDPF. These findings indicate that TLR2, TLR4, NOD2, and NOD1 in HDPF are functional receptors, and NOD2 is a modulator of signals transmitted through TLR2 in pulpal immune responses, leading to progressive pulpitis.
Yoshitaka Hosokawa, Ikuko Hosokawa, Kazumi Ozaki, Hideaki Nakae and Takashi Matsuo : Human gingival fibroblasts express functional chemokine receptor CXCR6., Clinical and Experimental Immunology, Vol.156, No.3, 413-418, 2009.
(要約)
We have reported that CXCL16, a recently discovered transmembrane chemokine, is expressed in human gingival fibroblasts (HGF). However, it is not known whether HGF express CXCR6, the receptor for CXCL16, or CXCL16 affects HGF biology. We have shown that HGF expressed CXCR6 by reverse transcription-polymerase chain reaction and flow cytometric analysis. Moreover, we elucidated that tumour necrosis factor (TNF)-alpha and cytosine-guanine dinucleotide (CpG) DNA (Toll-like receptor-9 ligand) treatment enhanced CXCR6 expression by HGF. Interleukin (IL)-4, IL-13 and CpG DNA up-regulated CXCR6 expression by TNF-alpha-stimulated HGF. On the other hand, IL-1beta and interferon-gamma inhibited CXCR6 expression on TNF-alpha-treated HGF. CXCL16 treatment induced HGF proliferation and phosphorylation of extracellular regulated kinase (ERK) and protein kinase B (AKT) in HGF. In conclusion, HGF expressed CXCR6 functionally, because CXCL16 induced HGF proliferation and ERK and AKT phosphorylation in HGF. These results indicate that CXCL16 may play an important role in the pathogenesis and remodelling in periodontally diseased tissues.
Yoshitaka Hosokawa, Ikuko Hosokawa, Kazumi Ozaki, Hideaki Nakae and Takashi Matsuo : Cytokines differentially regulate CXCL10 production by interferon-gamma-stimulated or tumor necrosis factor-alpha-stimulated human gingival fibroblasts., Journal of Periodontal Research, Vol.44, No.2, 225-231, 2009.
(要約)
CXC chemokine 10 (CXCL10) activates CXC chemokine receptor 3 (CXCR3) and attracts activated T-helper 1 cells. In this study we examined the effects of cytokines on CXCL10 production by human gingival fibroblasts. Human gingival fibroblasts were exposed to pro-inflammatory cytokines (interleukin-1beta, tumor necrosis factor-alpha), a T-helper 1 cytokine (interferon-gamma), T-helper 2 cytokines (interleukin-4, interleukin-13), T-helper 17 cytokines (interleukin-17A, interleukin-22) and regulatory T-cell cytokines (interleukin-10, transforming growth factor-beta1) for 24 h. CXCL10 production by human gingival fibroblasts was examined by enzyme-linked immunosorbent assay. Human gingival fibroblasts produced CXCL10 protein upon stimulation with interleukin-1beta, tumor necrosis factor-alpha and interferon-gamma. Treatment of human gingival fibroblasts with interferon-gamma in combination with tumor necrosis factor-alpha or interleukin-1beta resulted in a synergistic production of CXCL10. However, interleukin-4 and interleukin-13 inhibited CXCL10 production by interferon-gamma-stimulated or tumor necrosis factor-alpha-stimulated-human gingival fibroblasts. On the other hand, interleukin-17A and interleukin-22 enhanced CXCL10 production by human gingival fibroblasts treated with interferon-gamma and inhibited CXCL10 production by tumor necrosis factor-alpha-stimulated human gingival fibroblasts. Furthermore, the anti-inflammatory cytokine, interleukin-10, inhibited CXCL10 production by both interferon-gamma- and tumor necrosis factor-alpha-stimulated human gingival fibroblasts, but transforming growth factor-beta1 enhanced interferon-gamma-mediated CXCL10 production by human gingival fibroblasts. These results mean that the balance of cytokines in periodontally diseased tissue may be essential for the control of CXCL10 production by human gingival fibroblasts, and the production of CXCL10 might be important for the regulation of T-helper 1 cell infiltration in periodontally diseased tissue.
Liu Dali, Hiromichi Yumoto, Keiji Murakami, Katsuhiko Hirota, Shizuo Kayama, Taniguchi Tomonori, Yamamoto Akitake, Tsuneko Ono, Takashi Matsuo and Yoichiro Miyake : Heterologous expression of a histone-like protein from Streptococcus intermedius in Escherichia coli alters the nucleoid structure and inhibits the growth of E. coli, FEMS Microbiology Letters, Vol.288, No.1, 68-75, 2008.
(要約)
Escherichia coli failed to survive after transformation with a Streptococcus intermedius histone-like protein gene (Si-hlp) and its promoter-harbored plasmid. The promoter function of Si-hlp in E. coli was determined using enhanced green fluorescence protein (egfp) gene as a reporter. The inhibitory effect of Si-HLP on E. coli viability was verified by a tetracycline-inducible gene expression system. Further study suggested that Si-HLP may alter the bacterial nucleoid structure, leading to the growth inhibition of E. coli.
Kanako Takahashi, Tadashi Nakanishi, Hiromichi Yumoto, Tomohiko Adachi and Takashi Matsuo : CCL20 production is induced in human dental pulp upon stimulation by Streptococcus mutans and pro-inflammatory cytokines, Oral Microbiology and Immunology, Vol.23, No.4, 320-327, 2008.
(要約)
Pulpitis is characterized by the marked infiltration of inflammatory cells in response to an invasion of caries-related bacteria. It is well known that chemokines regulate the trafficking of lymphocytes, and CC chemokine ligand 20 (CCL20) has been recently shown to play a crucial role in the recruitment of memory T cells and immature dendritic cells into inflammatory lesions. We previously reported that CCL20 was mainly expressed in microvascular endothelial cells and macrophages that accumulated in inflamed pulp tissues and that its specific receptor, CCR6, was expressed on infiltrated lymphocytes. However, the mechanism of CCL20 expression remains unclear. In this study, we investigated the expression of CCL20 in monocytes/macrophages, endothelial cells, and pulpal fibroblasts after stimulation with Streptococcus mutans, a representative of caries-related bacteria, or proinflammatory cytokines. CCL20 messenger RNA was detected by reverse transcription-polymerase chain reaction in inflamed pulp, but not in clinically normal pulp. By enzyme-linked immunosorbent assay, S. mutans induced a human monocytic cell line, differentiated macrophage-like THP-1 cells, and human umbilical vein endothelial cells (HUVEC) to produce an increased amount of CCL20. Lipoteichoic acid from S. mutans also elicited CCL20 production by HUVEC. Moreover, CCL20 production from pulpal fibroblasts was increased by stimulation with inetrleukin-1beta and tumor necrosis factor-alpha. Our results indicate that CCL20 expression is induced by stimulation with caries-related bacteria that have invaded deeply into the dentinal tubules as well as by proinflammatory cytokines in the inflamed pulpal lesions. It may be involved in the progression of pulpitis via accumulation of inflammatory cells.
Yoshitaka Hosokawa, Ikuko Hosokawa, Kazumi Ozaki, Hideaki Nakae and Takashi Matsuo : CC Chemokine ligand 17 in periodontal diseases: expression in diseased tissues and production by human gingival fibroblasts, Journal of Periodontal Research, Vol.43, No.4, 471-477, 2008.
(要約)
It has been reported that T helper 2 (Th2) cells are related to exacerbation of periodontal disease. However, it is uncertain how the migration of Th2 cells is controlled. In this study, we examined the expression of CC chemokine ligand 17 (CCL17), which is a Th2 chemokine, in periodontal tissues. Moreover, we investigated the effects of cytokines and toll-like receptor (TLR) ligands on the production of CCL17 by human gingival fibroblasts (HGFs). We used immunohistochemistry and reverse transcriptase-polymerase chain reaction (RT-PCR) to detect CCL17 in periodontal tissues. HGFs were exposed to cytokines and TLR ligands. Expression of CCL17 was examined by RT-PCR and enzyme-linked immunosorbent assay. We used signal transduction inhibitors in some experiments. Both CCL17 and its receptor, CC chemokine receptor 4 (CCR4), were expressed in diseased periodontal tissues. A combination of tumour necrosis factor alpha (TNF-alpha) and interleukin (IL)-4/IL-13 increased CCL17 expression. Moreover, treatment of HGFs with a low dose of interferon-gamma (IFN-gamma) in combination with TNF-alpha and IL-4 or IL-13 had synergistic effects on the production of CCL17, whereas a high dose of IFN-gamma inhibited CCL17 production. Furthermore, Escherichia coli (E. coli) lipopolysaccharide (TLR4 ligand) and Pam3CSK4 (TLR2 ligand) inhibited CCL17 production by TNF-alpha + IL-4-stimulated HGFs, while CpG DNA (TLR9 ligand) enhanced TNF-alpha + IL-4 induced-CCL17 production by HGFs. Furthermore, a c-Jun NH2 terminal kinase (JNK) inhibitor, a phosphatidylinositol-3-kinase (PI3K) inhibitor and a nuclear factor kappa B (NF-kappa B) inhibitor inhibited CCL17 production by HGFs. These results suggest that the CCL17 produced by HGFs may be involved in the migration of Th2 cells into inflamed tissues, and provide evidence that CCL17 production is controlled by cytokines and TLR ligands in periodontal disease.
Ikuko Hosokawa, Yoshitaka Hosokawa, Kazumi Ozaki, Hideaki Nakae and Takashi Matsuo : Adrenomedullin suppresses tumour necrosis factor alpha-induced CXC chemokine ligand 10 production by human gingival fibroblasts., Clinical and Experimental Immunology, Vol.152, No.3, 568-575, 2008.
(要約)
Periodontal disease is an inflammatory disorder characterized by the involvement of chemokines that are important for the recruitment of leucocytes. Several cytokines, including tumour necrosis factor alpha (TNF-alpha), are involved in regulating levels of chemokines in periodontal disease. CXC chemokine ligand 10 (CXCL10) is a chemokine related to the migration of T helper 1 cells. In this study, we examined CXCL10 expression in human gingival fibroblasts (HGFs). Moreover, we investigated the effects of adrenomedullin (AM), which is a multi-functional regulatory peptide, on the production of CXCL10 by HGFs. We revealed that TNF-alpha stimulation induced CXCL10 production by HGFs. HGFs expressed AM and AM receptors, calcitonin-receptor-like receptor (CRLR) and receptor-activity-modifying protein (RAMP) 2, mRNAs constitutively. AM treatment supressed CXCL10 production by TNF-alpha-stimulated HGFs. Moreover, we elucidated that AM produced by HGFs inhibited CXCL10 production by HGFs, because AM antagonist enhanced CXCL10 production by HGFs. TNF-alpha treatment enhanced CRLR and RAMP2 mRNA expression in HGFs. Furthermore, AM is expressed in human periodontal tissues, including both inflamed and clinically healthy tissues. These results suggest that the CXCL10 produced by HGFs may be involved in the migration of leucocytes into inflamed tissues and related to exacerbation of periodontal disease. AM might be a therapeutic target of periodontal disease, because AM can inhibit CXCL10 production by HGFs.
Liu Dali, Hiromichi Yumoto, Keiji Murakami, Katsuhiko Hirota, Tsuneko Ono, Hideaki Nagamune, Shizuo Kayama, Takashi Matsuo and Yoichiro Miyake : The essentiality and involvement of Streptococcus intermedius histone-like DNA-binding protein in bacterial viability and normal growth, Molecular Microbiology, Vol.68, No.5, 1268-1282, 2008.
(要約)
Streptococcus intermedius histone-like DNA-binding protein (Si-HLP) is a homodimeric protein and, conserved with Escherichia coli HU, a well-documented nucleoid-associated protein (NAP). In E. coli, HU plays important roles as both structural and regulatory factors, but it is not essential for E. coli viability. Streptococcal HLP has been found to bind host cells and induce cytokine production, but its physiological role remains poorly defined. In the present study, using gene insertion knockout and tetracycline-regulated antisense RNA expression techniques, we determined whether Si-HLP is essential for bacterial viability and normal growth in S. intermedius. The Si-HLP-downregulated S. intermedius strain showed alterations in its morphology and surface properties. Downregulation of Si-HLP led to an expanded nucleoid to fill the intracellular space. Transcription levels of several genes, including virulence-associated factors, were found to be activated or repressed in the antisense Si-hlp RNA-expressing strain by real-time PCR and reverse-transcription PCR. Collectively, these data suggest that Si-HLP serves as an essential NAP governing the nucleoid architecture and controlling the gene transcription profile in S. intermedius.
Two days training camp for the fourth year students of dental school was executed in 2006 and 2007.The purpose of this program is to develop good practical dental doctors through the promotion ofcommunication skills. The training camp was held at the YMCA oceanic center in Anan city of TokushimaPrefecture. It began with an oceanic program under relaxed atmosphere, and then, the programscontaining games, lectures, and workshops related to communication skills were carried out. The theme "support for career formation as a medical profession" was added to "promotion of communication skills" in 2007,and then the skills to learn the definition and the practice of the medical team treatment were installed andexecuted.The usefulness of methodology different from the regular course class was verified based on the results,and the possibility of the application in the future curriculum for the career formation was discussed in thepaper.平成18 年度と19 年度に，歯学部4年生に対して1泊2日の合宿研修を正課外授業として実施した．本事業は，コミュニケーション能力の開発を通じてより良き医療人を育成することが目的である．合宿研修場所は徳島県内の海洋センターであった．リラックスした雰囲気で海洋プログラムからスタートし，次いで，コミュニケーションに対する導入的なゲームや講演，そしてワークショップにより自分たち自身で問題点を抽出し，さらに解決するための討論を行い，その結果をプレゼンテーションをするという内容であった．平成19 年度の2回目には，医学部と薬学部の協力を得て，実施テーマとして「コミュニケーション能力の育成」に「医療専門職としてのキャリア形成支援」という新たなテーマを加え，チーム医療の定義や実践を学ぶ機会を設けて実施した．従来の正課授業と異なった教育方法の可能性について成果を基に検証し，今後のキャリア形成支援教育への応用の可能性について考察した．
Dali Liu, Hiromichi Yumoto, Katsuhiko Hirota, Keiji Murakami, Kanako Takahashi, Kouji Hirao, Takashi Matsuo, Kazuto Ohkura, Hideaki Nagamune and Yoichiro Miyake : Histone-like DNA binding protein of Streptococcus intermedius induces the expression of pro-inflammatory cytokines in human monocytes via action of ERK1/2 and JNK pathways, Cellular Microbiology, Vol.10, No.1, 262-276, 2008.
(要約)
Streptococcus intermedius is a commensal associated with serious, deep-seated purulent infections in major organs, such as the brain and liver. Histone-like DNA binding protein (HLP) is an accessory architectural protein in a variety of bacterial cellular processes. In this study, we investigated the mechanisms of pro-inflammatory cytokine inductions in THP-1 cells by stimulation with recombinant HLP of S. intermedius (rSi-HLP). rSi-HLP stimulation-induced production of pro-inflammatory cytokines (IL-8, IL-1 beta and TNF-alpha) occurred in a time- and dose-dependent manner. In contrast with the heat-stable activity of DNA binding, the induction activity of rSi-HLP was heat-unstable. In subsequent studies, rSi-HLP acted cooperatively with lipoteichoic acid, the synthetic Toll-like receptor 2 agonist, Pam3CSK4, and the cytosolic nucleotide binding oligomerization domain 2 receptor agonist, muramyldipeptide. Furthermore, Western blot and blocking assays with specific inhibitors showed that rSi-HLP stimulation induced the activation of cell signal transduction pathways, extracellular signal-regulated kinase 1/2 (ERK1/2) and c-Jun N-terminal kinase (JNK). In addition to its physiological role in bacterial growth through DNA binding, these results indicate that Si-HLP can trigger a cascade of events that induce pro-inflammatory responses via ERK1/2 and JNK signal pathways, and suggest that bacterial HLP may contribute to the activation of host innate immunity during bacterial infection.
(キーワード)
Acetylmuramyl-Alanyl-Isoglutamine / Bacterial Proteins / Cell Line / Cytokines / DNA, Bacterial / DNA-Binding Proteins / Hot Temperature / Humans / JNK Mitogen-Activated Protein Kinases / Lipopolysaccharides / Mitogen-Activated Protein Kinase 3 / Molecular Sequence Data / Monocytes / Recombinant Proteins / Sequence Analysis, DNA / Signal Transduction / Streptococcus intermedius / Teichoic Acids / Up-Regulation
Tomohiko Adachi, Tadashi Nakanishi, Hiromichi Yumoto, Kouji Hirao, Kanako Takahashi, Kayo Mukai, Hideaki Nakae and Takashi Matsuo : Caries-related bacteria and cytokines induce CXCL10 in dental pulp., Journal of Dental Research, Vol.86, No.12, 1217-1222, 2007.
(要約)
Marked infiltration of inflammatory cells, such as activated T-cells, is observed in the progression of pulpitis; however, little is known about the mechanism of their recruitment into pulpal lesions. It has been recently demonstrated that CXC chemokine ligand 10 (CXCL10) chemoattracts CXC chemokine receptor 3 (CXCR3)-positive activated T-cells. We therefore examined whether CXCL10 is involved in the pathogenesis of pulpitis. CXCL10 mRNA expression levels in clinically inflamed dental pulp were higher than those in healthy dental pulp. Immunostaining results revealed that CXCL10 was detected in macrophages, endothelial cells, and fibroblasts in inflamed dental pulp, and that CXCR3 expression was observed mainly on T-cells. Moreover, cultured dental pulp fibroblasts produced CXCL10 after stimulation with live caries-related bacteria, peptidoglycans, and pro-inflammatory cytokines. In contrast, heat-killed bacteria did not induce CXCL10 secretion. These findings suggest that CXCL10-CXCR3 may play an important role in the pulpal immune response to caries-related bacterial invasion. Abbreviations: CXCL10, CXC chemokine ligand 10; CXCR3, CXC chemokine receptor 3; IFN, interferon; FBS, fetal bovine serum; LTA, lipoteichoic acid; PGN, peptidoglycan; IL, interleukin; TNF, tumor necrosis factor; PBS, phosphate-buffered saline; ELISA, enzyme-linked immunosorbent assay; CCL, C-C chemokine ligand; TLR, Toll-like receptor; NOD, nucleotide oligomerization domain; HDPF, human dental pulp fibroblasts.
Chihiro Shinohara, Kikuji Yamashita, Takashi Matsuo, Seiichiro Kitamura and Fumiaki Kawano : Effects of Carbonic AnhydraseInhibitor Acetazolamide(AZ)on Osteoclasts and Bone Structure, Journal of Hard Tissue Biology, Vol.16, No.3, 115-123, 2007.
(要約)
We examined the changes of osteoclasts and bone structure in vivo and in vitro under the condition of low activity of bone resorption induced by acetazolamide (AZ). AZ is a specific inhibitor of carbonic anhydrase II (CA II), which is the enzyme indispensable for substantial proton generation by osteoclasts. In vivo study, we investigated histologically and histomorphometrically the changes of trabecular bone and osteoclasts at the cartilage-bone junction of tibiae in rats injected subcutaneously with AZ for 21 days. The area of trabecular bone of AZ treated group increased compared with control group. Moreover, AZ treatment decreased about 20% the number of osteoclasts at the cartilage-bone junction. In vitro study, we examined the effects of AZ on osteoclasts using culture system of rat bone marrow. The number of osteoclasts on the bone slice was suppressed by AZ of a concentration from 10^<-5> to 10^<-4> M in a doze-dependent manner. The number of osteoclasts cultured with 10^<-4>M AZ for 24 hours and with or without parathyroid hormone (PTH) on the bone slice and on the glass showed a significant decrease compared with the control group. Some osteoclasts treated with AZ showed morphological changes of apoptosis-like cell death, including shrinking of cytoplasm and fragmentation of nuclei. These results show that high-doze of AZ decreases bone resorption not only by inhibiting CA II but also by decreasing the number of osteoclasts via induction of cell death. This suggests that CA II plays an important role not only in proton formation but also in osteoclast survival.
(キーワード)
Acetazolamide / Bone / Carbonic anhydrase II / Osteoclast
Taro Muto, Keiko Miyoshi, Seiichi Munesue, Hiroshi Nakada, Minoru Okayama, Takashi Matsuo and Takafumi Noma : Differential expression of syndecan isoforms during mouse incisor amelogenesis., The Journal of Medical Investigation : JMI, Vol.54, No.3-4, 331-339, 2007.
(要約)
Syndecans are transmembranous heparan sulfate proteoglycans (HSPGs) with covalently attached glycosaminoglycan side-chains located on the cell surface. The mammalian syndecan family is composed of four types of syndecans (syndecan-1 to -4). Syndecans interact with the intracellular cytoskeleton through the cytoplasmic domains of their core proteins and membrane proteins, extracellular enzymes, growth factors, and matrix components, through their heparan-sulfate chains, to regulate developmental processes.Here, as a first step to assess the possible roles of syndecan proteins in amelogenesis, we examined the expression patterns of all syndecan isoforms in continuously growing mouse incisors, in which we can overview major differentiation stages of amelogenesis at a glance. Understanding the expression domain of each syndecan isoform during specific developmental stages seems useful for investigating their physiological roles in amelogenesis. Immunohistochemical analysis of syndecan core proteins in the lower incisors from postnatal day 1 mice revealed spatially and temporally specific expression patterns, with syndecan-1 expressed in undifferentiated epithelial and mesenchymal cells, and syndecan-2, -3, and -4 in more differentiated cells. These findings suggest that each syndecan isoform functions distinctly during the amelogenesis of the incisors of mice.
Yoshitaka Hosokawa, Ikuko Hosokawa, Kazumi Ozaki, Hideaki Nakae and Takashi Matsuo : CXC chemokine ligand 16 in periodontal diseases: expression in diseased tissues and production by cytokine-stimulated human gingival fibroblasts., Clinical and Experimental Immunology, Vol.149, No.1, 146-154, 2007.
(要約)
Periodontal disease is an inflammatory disorder characterized by the involvement of chemokines that are important for the recruitment of leucocytes. Several cytokines are involved in regulating levels of chemokines in periodontal disease. CXCL16 is a chemokine related to the migration of T helper 1 (Th1) cells and natural killer (NK) cells. In this study, we examined its expression in periodontal tissues. Moreover, we investigated the effects of cytokines on the production of CXCL16 by human gingival fibroblast (HGF). Reverse transcription-polymerase chain reaction (RT-PCR) analysis and immunohistochemistry revealed that CXCL16 and its receptor, CXCR6, were expressed at the mRNA and protein levels in diseased tissues. Proinflammatory cytokines [interleukin (IL)-1beta, tumour necrosis factor (TNF)-alpha and interferon (IFN)-gamma] increased the mRNA expression and release of CXCL16 in a dose-dependent manner. Moreover, treatment of HGFs with IFN-gamma in combination with IL-1beta had a synergistic effect on the production of CXCL16. On the other hand, IL-4 and IL-13 inhibited the IL-1beta-induced CXCL16 production by HGFs. Inhibitors of A disintegrin and metalloprotease (ADAM)10 and ADAM17, a recently identified protease of CXCL16, reduced the amount of CXCL16 released from HGFs. These results suggest that the CXCL16 produced by HGFs may be involved in the migration of leucocytes into inflamed tissues, and provide evidence that CXCL16 production is controlled by cytokines in periodontal disease.
Hiromichi Yumoto, Masayoshi Yamada, Chihiro Shinohara, Hideaki Nakae, Kanako Takahashi, Hiroyuki Azakami, Ebisu Shigeyuki and Takashi Matsuo : Soluble products from Eikenella corrodens induce cell proliferation and expression of interleukin-8 and adhesion molecules in endothelial cells via mitogen-activated protein kinase pathways, Oral Microbiology and Immunology, Vol.22, No.1, 36-45, 2007.
(要約)
The periodontal vasculature is profoundly affected during the progression of periodontitis, and several specific bacteria are believed to be involved in this inflammatory disease. Eikenella corrodens is one of the common bacteria detected in periodontitis diseased lesions; however, the function of this organism in periodontitis is not well understood. In this study, we investigated the E. corrodens-induced endothelial cell alteration and inflammation process that leads to leukocyte infiltration in inflamed regions. Soluble products from E. corrodens (EcSP) induced the gene expression and protein production of vascular endothelial growth factor in oral epithelial cells and human umbilical vein endothelial cells (HUVEC). Direct stimulation by EcSP also activated endothelial cell proliferation. Moreover, EcSP induced ERK1/2 (p44/42) and p38 mitogen-activated protein kinase (MAPK) phosphorylation within 10-30 min in HUVEC, as demonstrated by Western blot analysis and up-regulated intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1), E-selectin and interleukin-8 (IL-8) production demonstrated by reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay. The specific p38 MAPK inhibitor SB203580 reduced the expression of ICAM-1, VCAM-1 and IL-8, whereas the blockade of p44/42 by MAPK kinase (MEK1) inhibitor, PD98059, inhibited only IL-8 expression. Our results indicate that E. corrodens can trigger a cascade of events that induce inflammatory responses in periodontal tissue via the MAPK cascade and may promote chronic periodontitis without bacteria-cell contact.
Yoshitaka Hosokawa, Ikuko Hosokawa, Kazumi Ozaki, Hideaki Nakae and Takashi Matsuo : Proinflammatory effects of tumour necrosis factor-like weak inducer of apoptosis (TWEAK) on human gingival fibroblasts., Clinical and Experimental Immunology, Vol.146, No.3, 540-549, 2006.
(要約)
Tumour necrosis factor (TNF)-like weak inducer of apoptosis (TWEAK), a member of the TNF family, is a multi-functional cytokine that regulates cellular proliferation, angiogenesis, inflammation and apoptosis. In this study, we investigated TWEAK expression in periodontally diseased tissues and the effect of TWEAK on human gingival fibroblasts (HGF). Reverse transcription-polymerase chain reaction (RT-PCR) analysis and immunohistochemistry revealed that TWEAK and the TWEAK receptor, fibroblast growth factor-inducible 14 (Fn14), mRNA and protein were expressed in periodontally diseased tissues. HGF expressed Fn14 and produced interleukin (IL)-8 and vascular endothelial growth factor (VEGF) production upon TWEAK stimulation in a dose-dependent manner. The IL-8 and VEGF production induced by TWEAK was augmented synergistically by simultaneous stimulation with transforming growth factor (TGF)-beta1 or IL-1beta. IL-1beta and TGF-beta1 enhanced Fn14 expression in a dose-dependent manner. Moreover, TWEAK induced intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) expression on HGF in a dose-dependent manner. The ICAM-1 expression induced by TWEAK was augmented by TGF-beta1. On the other hand, the TWEAK-induced VCAM-1 expression was inhibited by TGF-beta1. Phosphatidylinositol 3-kinase (PI3K) and nuclear factor-kappaB (NF-kappaB) inhibitor inhibit both ICAM-1 and VCAM-1 expression induced by TWEAK. However, mitogen-activated protein kinase (MEK) and c-Jun NH2-terminal kinase (JNK) inhibitor enhanced only VCAM-1 expression on HGF. These results suggest that TWEAK may be involved in the pathophysiology of periodontal disease. Moreover, in combination with IL-1beta or TGF-beta1, TWEAK may be related to the exacerbation of periodontal disease to induce proinflammatory cytokines and adherent molecules by HGF.
Yoshitaka Hosokawa, Ikuko Hosokawa, Kazumi Ozaki, Hideaki Nakae and Takashi Matsuo : Cytokines differentially regulate ICAM-1 and VCAM-1 expression on human gingival fibroblasts., Clinical and Experimental Immunology, Vol.144, No.3, 494-502, 2006.
(要約)
The expression of intercellular adhesion molecule-1 (ICAM-1) and vascular adhesion molecule-1 (VCAM-1) on human gingival fibroblasts (HGF) may be important for migration and retention of inflammatory cells in periodontally diseased tissue. This study aimed to assess which cytokines regulate ICAM-1 and VCAM-1 expression on HGF. Tumour necrosis factor (TNF)-alpha and interferon (IFN)-gamma enhanced both ICAM-1 and VCAM-1 expression on HGF. Interleukin (IL)-1beta mainly up-regulated ICAM-1 expression. On the other hand, IL-4 and IL-13 enhanced only VCAM-1 expression on HGF. IL-10 did not modulate both ICAM-1 and VCAM-1 expression. Transforming growth factor (TGF)-beta1 enhanced ICAM-1 expression. However, TGF-beta1 inhibited the VCAM-1 expression induced by TNF-alpha or IL-4. Both ICAM-1 and VCAM-1 expression by HGF was inhibited by nuclear factor-kappaB (NF-kappaB) activation inhibitor (MG-132). Mitogen-activated protein kinases (MAPK) inhibitors did not influence ICAM-1 expression induced by TNF-alpha. Interestingly, VCAM-1 expression was enhanced by MEK inhibitor (PD98059) and c-Jun NH2-terminal kinase (JNK) inhibitor (SP600125). These results mean that the balance of cytokines in periodontally diseased tissue may be essential for control of ICAM-1 and VCAM-1 expression on HGF, and the balance of ICAM-1 and VCAM-1 expression might be important for regulation of leucocytes infiltration and retention in periodontally diseased tissue.
Toshiyuki Suge, Akiko Kawasaki, Kunio Ishikawa, Takashi Matsuo and Shigeyuki Ebisu : Effect of ammonium hexafluorosilicate on dentin tubule occlusion for the treatment of dentin hypersensitivity, American Journal of Dentistry, Vol.19, No.4, 248-252, 2006.
(要約)
To evaluate the occluding ability of ammonium hexafluorosilicate (SiF). Dentin disks prepared from human extracted teeth were grouped as follows to prepare different situations of dentin hypersensitivity: (1) those sonicated for 20 minutes; (2) those treated with 0.5 mol/L EDTA for 2 minutes; (3) those treated with 6% citric acid for 2 minutes; (4) those treated with 50% citric acid for 2 minutes. Then, SiF or diamine silver fluoride (AgF) was applied to the dentin disks and the dentin tubule occlusion was observed with scanning electron microscopy (SEM). The percent of open tubules before and after SiF or AgF treatment were measured by NIH image using SEM photographs. Also, the dentin permeability was measured. SEM micrographs demonstrated that the dentin tubules were completely occluded by the precipitate after SiF treatment. Also, the dentin permeability was reduced to 10.3%. In contrast, most of the dentin tubules remained open after AgF treatment. EDXA analysis showed that the precipitate in the dentin tubules that forms after SiF treatment contains Si, Ca and P, indicating a silica-calcium phosphate complex.
Toshiyuki Suge, Akiko Kawasaki, Kunio Ishikawa, Takashi Matsuo and Shigeyuki Ebisu : Effects of plaque control on the patency of dentinal tubules-an in vivo study in beagle dogs, Journal of Periodontology, Vol.77, No.3, 454-459, 2006.
(要約)
The aim of this in vivo study was to evaluate the effects of plaque control on the patency of dentinal tubules using vital teeth of beagle dogs. Class V cavities were prepared on the cervical areas of the mandibular and maxillary molars in each dog with a diamond point. To simulate the state of dentinal hypersensitivity, the teeth were etched with 50% citric acid for 2 minutes to obtain patent dentinal tubules. Plaque control was achieved by brushing the left-side teeth every day, whereas no plaque control was performed for the right-side teeth. A dentin biopsy was performed after 1, 2, and 3 weeks using the cylindrical diamond point to obtain dentin specimens. In the plaque control group, some of the dentinal tubules were occluded with precipitate (Ca/P=1.49), and the diameter of the dentinal tubules decreased from 2.42+/-0.33 microm (mean+/-SD) to 1.11+/-0.51 microm after 7 days, although most of the dentinal tubules remained open. In contrast, no precipitate was observed in the dentinal tubules of the non-plaque control group. Also, the diameter of the dentinal tubules increased from 2.42+/-0.33 to 2.9+/-0.49 microm, due to the demineralization of the peritubular and intertubular dentin. Plaque control plays a key role in reducing the patency of dentinal tubules and, therefore, might promote the natural repair of dentinal hypersensitivity.
Toshiyuki Suge, Akiko Kawasaki, Kunio Ishikawa, Takashi Matsuo and SHigeyuki Ebisu : Comparison of the occluding ability of dentinal tubules with different morphology between calcium phosphate precipitation method and potassium oxalate treatment, Dental Materials Journal, Vol.24, No.4, 522-529, 2005.
(要約)
The aim of this study was to evaluate the occluding ability of calcium phosphate precipitation (CPP) method and potassium oxalate treatment when each method was applied to dentin disks with different surface morphology. Occluding ability was observed by scanning electron microscopy. Irrespective of the diameter of the dentinal tubules, the CPP method showed a consistent occluding ability for dentinal tubules at the dentin surface, and that the depths of the precipitate formed in the dentinal tubules by CPP method were not significantly different. In contrast, the occluding ability of potassium oxalate treatment was reduced with increasing diameter of the dentinal tubules. However, the reduction of the occluding ability of potassium oxalate treatment was more markedly affected by the demineralization of dentin surface. Since the CPP method showed a consistent occluding ability irrespective of the diameter of the dentinal tubules, it is suggested that the CPP method would be a useful means for treating dentin hypersensitivity.
Yoshitaka Hosokawa, Ikuko Hosokawa, Kazumi Ozaki, Hideaki Nakae and Takashi Matsuo : Increase of CCL20 expression by human gingival fibroblasts upon stimulation with cytokines and bacterial endotoxin, Clinical and Experimental Immunology, Vol.142, No.2, 285-291, 2005.
(要約)
We have demonstrated recently that CCL20 was expressed in periodontal diseased tissues and abundant CCR6 positive T cells infiltrated in periodontally diseased tissue. However, it is uncertain which cells can elicit CCL20 production. In the present study, we examined the properties of CCL20 production by human gingival fibroblasts (HGF) culture. Here, we report that interleukin-1 beta (IL-1beta), tumour necrosis factor-alpha (TNF-alpha) and Escherichia coli lipopolysaccharide (LPS) can significantly induce the production of CCL20 by HGF. We found that TNF-alpha and E. coli LPS enhanced the production of CCL20 by HGF treated with IL-1beta. In contrast, interferon-gamma (IFN-gamma) dramatically diminished CCL20 production induced by IL-1beta. Moreover, we demonstrated that nuclear factor-kappaB (NF-kappaB), p38 mitogen-activated protein kinase (MAPK) and extracellular signal-regulated kinases (ERK) play an important role in mediating the production of CCL20 induced by IL-1beta and TNF-alpha. On the other hand, we found that not only NF-kappaB, p38 MAPK and ERK but also c-Jun NH2-terminal kinase (JNK) are involved in CCL20 production induced by E. coli LPS. Finally, we found that HGF express CCR6, CCL20 receptor, and CCL20 induced vascular endothelial growth factor (VEGF) by HGF. Taken together, these findings that HGF will be a source of CCL20 in periodontal tissue, and the CCL20 production will be controlled by proinflammatory cytokine and bacterial LPS in periodontally diseased tissue. Thus, CCL20 by HGF might be involved in inflammatory cells infiltration, and promote the progression of periodontal disease.
Yoshitaka Hosokawa, Ikuko Hosokawa, Kazumi Ozaki, Hideaki Nakae, Keiji Murakami, Yoichiro Miyake and Takashi Matsuo : CXCL12 and CXCR4 expression by human gingival fibroblasts in periodontal disease, Clinical and Experimental Immunology, Vol.141, No.3, 467-474, 2005.
(要約)
CXCL12 is a CXC chemokine that is related to lymphocyte infiltration and angiogenesis in inflammatory sites such as arthritis. However, the expression and roles of CXCL12 in periodontal disease are uncertain. The aim of this study was to assess the expression of CXCL12 and its receptor, CXCR4, in periodontal tissue and to investigate the properties of CXCL12 and CXCR4 expression by human gingival fibroblasts (HGF). RT-PCR analysis revealed that CXCL12 and CXCR4 mRNA were expressed in both normal gingival tissues and periodontal diseased tissues. Immunohistochemistry disclosed that CXCL12 was expressed and CXCR4 positive cells were found in both normal and periodontal diseased gingival tissues. Our in vitro experiments elucidated that HGF constitutively produced CXCL12, and the levels were enhanced by stimulation with tumour necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma), transforming growth factor-beta (TGF-beta), regulated upon activation normal T cell expressed and secreted (RANTES) and macrophage inflammatory protein 3(alpha) (MIP-3(alpha)). On the other hand, heat killed Porphyromonas gingivalis (P. gingivalis) and P. gingivalis LPS reduced the CXCL12 production by HGF. Flow cytometry analysis clarified that CXCR4 was highly expressed on HGF, and CXCR4 expression was abrogated by TNF-alpha, IFN-gamma and P. gingivalis LPS. Moreover, CXCL12 induced vascular endothelial growth factor (VEGF) production by HGF. Our results demonstrated that CXCL12 might be related to CXCR4+ cells infiltration and angiogenesis both in normal periodontal tissues and periodontal diseased tissue. P. gingivalis, a known periodontal pathogen, inhibits the production of CXCL12 and the expression of CXCR4 by HGF. This fact means that P. gingivalis may inhibit CXCR4+ cells infiltration and neovascularization in periodontal tissue and escape from the immune response.
Akiko Kawasaki, Toshiyuki Suge, Kunio Ishikawa, Kazumi Ozaki, Takashi Matsuo and Shigeyuki Ebisu : Ammonium hexafluorosilicate increase acid resistance of bovine enamel and dentine, Journal of Materials Science. Materials in Medicine, Vol.16, No.5, 461-466, 2005.
(要約)
Although diamine silver fluoride (AgF: (NH3)2AgF) stains teeth black, it is known as a very effective agent to prevent the dental caries progress. In order to find another fluoride that has a similar anticariogenic effect without changing tooth color, we prepared ammonium hexafluorosilicate (SiF: (NH4)2SiF6), in which the silver of AgF is replaced with silicon. In this study, the anticariogenic effect of SiF was evaluated using bovine teeth. Fluoride solutions, SiF, AgF, acidulated phosphate fluoride (APF), and sodium fluoride (NaF), were applied to bovine enamel and dentine blocks, and the depth of demineralization was measured after exposure to a demineralizing solution for 24 h. Also, fluoride was applied to a simulated dentine caries specimen to evaluate the caries progress-preventing ability. For the dentine specimens, mineral loss (Delta Z) was also measured with microradiography. We found that SiF treated enamel showed better acid resistance than specimens treated with NaF or APF. AgF treated enamel also showed similar acid resistance, but was stained black. SiF and AgF treated caries-affected dentine showed reduced demineralization when exposed to a demineralization solution for 24 h. Mineral loss (Delta Z) was reduced to 85% and 75%, respectively. Although the acid resistance of the SiF treated teeth was inferior to that of the AgF treated teeth, we consider that SiF has good potential as anticariogenic agent, since it increased acid resistance without changing tooth color.
Toshiyuki Suge, Ishikawa Kunio, Akiko Kawasaki, Takashi Matsuo and EBISU Shigeyuki : Effects of pre- or post- application with calcium chloride on occluding ability of potassium oxalate for the treatment of dentin hypersensitivity, American Journal of Dentistry, Vol.18, No.2, 121-125, 2005.
(要約)
To evaluate whether calcium ion supply using 1-6 mol/L CaCl2 solution could enhance the occluding ability of dentin tubules with 30% potassium oxalate treatment. Calcium chloride solution (1-6 mol/L) was applied to dentin disks before or after application of 30% potassium oxalate solution. Occluding ability after potassium oxalate treatment was evaluated with scanning electron microscopic (SEM) observation and measurement of dentin permeability. In addition, the composition of the precipitate formed when mixing potassium oxalate and calcium chloride was analyzed with a powder x-ray diffractometer (XRD). SEM observations revealed that the dentin tubules were occluded homogeneously and completely with the precipitate when calcium chloride solution was applied before or after potassium oxalate treatment. However, the depth of the precipitate in dentin tubules from the dentin surface became shallower when pre-treated with calcium chloride before potassium oxalate. Although dentin permeability was greatly reduced in both groups, no significant difference could be observed between samples with and without calcium chloride application.
Yoshitaka Hosokawa, Tadashi Nakanishi, Daisuke Yamaguchi, Hideaki Nakae and Takashi Matsuo : Expression of fractalkine (CX3CL1) and its receptor, CX3CR1, in periodontal diseased tissue, Clinical and Experimental Immunology, Vol.139, No.3, 506-512, 2005.
(要約)
The regulatory role of chemokines and chemokine receptors on specific leucocyte recruitment into periodontal diseased tissue is poorly characterized. We observed that leucocytes infiltrating inflamed gingival tissue expressed marked levels of CX3CR1. In periodontal diseased tissue, the expression of fractalkine and CX3CR1 mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR) and further, fractalkine was distributed mainly on endothelial cells, as shown by immunohistochemistry. Moreover, we can detect CX3CR1-expressing cells infiltrated in periodontal diseased tissue by immunohistochemical staining. Furthermore, fractalkine production by human umbilical vein endothelial cells (HUVEC) was up-regulated by pathogen-associated molecular patterns (PAMPs), including Porphyromonas gingivalis lipopolysaccharide (LPS). Thus, these findings suggested that CX3CR1 and the corresponding chemokine, fractalkine may have an important regulatory role on specific leucocyte migration into inflamed periodontal tissue.
Tadashi Nakanishi, Kanako Takahashi, Yoshitaka Hosokawa, Tomohiko Adachi, Hideaki Nakae and Takashi Matsuo : Expression of macrophage inflammatory protein 3alpha in human inflamed dental pulp tissue, Journal of Endodontics, Vol.31, No.2, 84-87, 2005.
(要約)
Severe pulpitis resulting from dental caries is characterized by marked inflammatory infiltrate such as lymphocytes. Little is known about the recruitment of these cells into the dental pulp lesions of carious teeth. Macrophage inflammatory protein-3alpha (MIP-3alpha), a CC chemokine attracts CC chemokine receptor 6 (CCR6)-expressing T cells. We examined the distribution of MIP-3alpha-positive and/or CCR6-positive cells in human inflamed and normal dental pulp by immunohistochemistry. MIP-3alpha was observed in all inflamed pulp sections, and was mostly distributed in macrophages that had accumulated in the area adjacent to carious lesions. Furthermore, CCR6 expression was also observed in the infiltrating lymphocytes. In contrast, MIP-3alpha and CCR6 were rarely detected in normal pulp. These findings suggest that MIP-3alpha plays a role in the advancement of pulpal inflammation via the recruitment of CCR6-expressing lymphocytes.
Takashi Matsuo, Toshiyuki Shirakami, Kazumi Ozaki, Tadashi Nakanishi, Hiromichi Yumoto and Shigeyuki Ebisu : An immunohistological study of the localization of bacteria invading root pulpal walls of teeth with periapical lesions, Journal of Endodontics, Vol.29, No.3, 194-200, 2003.
(要約)
We immunohistologically examined the prevalence and localization of bacteria invading dentinal tubules of the roots of teeth with infected canals. Forty extracted teeth with apical lesions were selected and divided into two groups: a group of untreated teeth and a group of canal-enlarged teeth. The bacteria in the specimens were detected by Brown-Brenn stain and the labeled-streptavidin-biotin method with specific antisera for 16-bacteria. Seventy percent of the examined teeth showed bacteria invading the dentinal tubules of the roots. Fusobacterium nucleatum, Eubacterium alactolyticum, E. nodatum, Lactobacillus casei, and Peptostreptococcus micros were abundant. Even in the canal-enlarged group, invasion of bacteria was observed in 65% of teeth. This study revealed the actual condition of bacteria in infected root dentin and suggested that the canal-enlargement procedure could not completely remove all the bacteria in the infected dentinal tubules of the root.
Masayoshi Yamada, Hideaki Nakae, Hiromichi Yumoto, Chihiro Shinohara, Shigeyuki Ebisu and Takashi Matsuo : N-acetyl-D-galactosamine specific lectin of Eikenella corrodens induces intercellular adhesion molecule-1 (ICAM-1) production by human oral epithelial cells, Journal of Medical Microbiology, Vol.51, No.12, 1080-1089, 2002.
(要約)
During the acute inflammatory response in periodontitis, gingival epithelial cells are considered to play important roles in the recruitment of inflammatory cells to the site of infection through the secretion of chemokines. However, little is known about the expression of molecules that are involved in the interaction between the epithelium and neutrophils following bacterial attachment. Earlier work reported that periodontopathogenic Eikenella corrodens strain 1,073 up-regulated the expression and secretion of chemokines such as interleukin-8 (IL-8) from KB cells (a human oral epithelial cell line derived from a human oral epidermoid carcinoma). To elucidate the mechanism of the transmigration of neutrophils through the epithelium, the present study investigated the expression of adhesion molecules on KB cells in response to E. corrodens attachment. Adhesion molecule gene expression was assessed by RT-PCR and adhesion proteins expressed on KB cell surfaces were determined by cell-based ELISA and FACS. In RT-PCR, ICAM-1 mRNA levels were significantly increased within 1 h in response to exposure to E. corrodens and continued to increase over the 12-h period of study. In ELISA, increased surface ICAM-1 expression was paralleled by increased ICAM-1 mRNA levels. Furthermore, the increases in ICAM-1 expression on epithelial cells infected with E. corrodens were observed to be due to the N-acetyl-D-galactosamine (GalNAc) specific bacterial lectin-like substance of E. corrodens (EcLS), which was one of the adhesins of E. corrodens. This is the first study to report that a bacterial lectin-like substance increased the expression of ICAM-1 on gingival epithelial cells.
(キーワード)
レクチン / ICAM-1 / Eikenella corrodens / 上皮細胞
(文献検索サイトへのリンク)
● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 12466406
Toshiyuki Suge, Kunio ISHIKAWA, Akiko Kawasaki, Kazuomi SUZUKI, Takashi Matsuo, Yuichiro NOIRI, Satoshi IMAZATO and Shigeyuki EBISU : Calcium phosphate precipitation method for the treatment of dentin hypersensitivity, American Journal of Dentistry, Vol.15, No.4, 220-226, 2002.
(要約)
To evaluate the feasibility of the calcium phosphate precipitation (CPP) method as a treatment for dentin hypersensitivity using vital teeth of beagle dogs. Also, gingival tissue response to two types of CPP methods was examined histologically. Dentin tubules of the dogs' vital teeth were exposed by shallow cavity preparation followed by etching with 50% citric acid to simulate the condition of hypersensitive dentin. After CPP treatment was applied to the vital tooth, the dentin surface and longitudinal sections were observed by SEM to evaluate the occluding ability of the CPP method. The precipitate was also analyzed by energy-dispersive X-ray microanalysis to obtain compositional information. The gingival tissue before and after CPP method was examined histologically with light microscopy. Dentin tubules were occluded homogeneously and completely with an apatitic mineral after application of the CPP treatment in vital teeth. However, the depth of the precipitate in dentin tubules from the dentin surface was approximately half that seen in extracted teeth. No histological change was observed in gingival tissues when NaHCO3 was used as a post-treatment solution of the CPP method, whereas another CPP method using NaOH solution resulted in atrophy and degeneration of the epithelium of gingival tissue.
Yoshitaka Hosokawa, Tadashi Nakanishi, Daisuke Yamaguchi, Kanako Takahashi, Hiromichi Yumoto, Kazumi Ozaki and Takashi Matsuo : Macrophage Inflammatory Protein 3α-CC chemokine receptor 6 interactions play an important role in CD4+ T-cell accumulation in periodontal diseased tissue., Clinical and Experimental Immunology, Vol.128, No.3, 548-554, 2002.
(要約)
The regulatory role of chemokines and chemokine receptors on specific lymphocyte recruitment into periodontal diseased tissue is poorly characterized. We observed that lymphocytes infiltrating inflamed gingival tissue expressed marked levels of CCR6. In periodontal diseased tissue, the expression of MIP-3alpha mRNA was detected by RT-PCR and further, MIP-3alpha was distributed in the basal layer of gingival epithelial cells, microvascular endothelial cells and the areas of inflammatory cells as shown by immunohistochemistry. Moreover, CCR6-expressing cells infiltrated into periodontal diseased tissue, and the proportion of CCR6-positive CD4+ T cells was significantly elevated in periodontal diseased tissue compared with peripheral blood in the same patients. Furthermore, gingival lymphocytes isolated from patients showed migration toward MIP-3alpha in an in vitro chemotaxis assay in which migration was abrogated by specific antibody to CCR6. Thus, these findings suggested that CCR6 and the corresponding chemokine, MIP-3alpha may have an important regulatory role in specific lymphocyte migration into inflamed periodontal tissue.
Hiromichi Yumoto, Hideaki Nakae, Masayoshi Yamada, Keiko Fujinaka, Chihiro Shinohara, Shigeyuki Ebisu and Takashi Matsuo : Soluble products from Eikenella corrodens stimulate oral epithelial cells to induce inflammatory mediators, Oral Microbiology and Immunology, Vol.16, No.5, 296-305, 2001.
(要約)
In the inflammatory response elicited by bacterial colonization in periodontal pockets, pocket epithelial cells not only serve as a barrier to isolate the pocket microenvironment from external stimuli but also regulate the functions of neighboring cells including fibroblasts and inflammatory cells. To elucidate this mechanism, we characterized the effects of periodontopathic bacterium Eikenella corrodens 1073 components on the production of some inflammatory mediators in a human oral epithelial cell line (KB). In enzyme-linked immunosorbent assay (ELISA), the E. corrodens supernatant induced interleukin-6 (IL-6), IL-8 and prostaglandin E2 but not interferon-gamma (IFN-gamma) production by KB cells. After incubation with E. corrodens supernatant, KB cells showed a marked increase in the levels of IL-6, IL-8 and PG G/H synthase (cyclooxygenase)-2, but not IFN-gamma, gene expression by reverse-transcriptase polymerase chain reaction. All these E. corrodens products responsible for production of these inflammatory mediators resisted freezing and boiling and were present in a 10-kDa filtrate. These results imply that these soluble small-molecular-mass products from E. corrodens stimulate various inflammatory mediator productions by human oral epithelial cells and may play a role in the initiation of periodontal inflammation and subsequently perpetuate the inflammatory response during chronic infection.
Tadashi Nakanishi, Hirotoshi Shimizu, Yoshitaka Hosokawa and Takashi Matsuo : An immunohistological study on cyclooxygenase-2 in human dental pulp., Journal of Endodontics, Vol.27, No.6, 385-388, 2001.
(要約)
Characteristics of cyclooxygenase-2 (COX-2) expressing cells in human dental pulp were immunohistologically studied. Extirpated pulpal tissues from extracted teeth were examined to elucidate the localization and distribution of COX-2. Pulpal tissues were examined by the labeled streptavidin biotin method using specific mouse monoclonal antibodies for COX-2. Cell types of the COX-2 expressing cells were also investigated by the double stain technique using both monoclonal antibodies for CD68/macrophage and anti-COX-2. COX-2 expressing cells could be found in all of the inflamed pulps, and these cells were mostly distributed close to the area of accumulation of inflammatory cells. COX-2 was mainly expressed in fibroblasts rather than macrophages. In contrast, COX-2 expressing cells were scarcely found in the normal pulps. These findings indicate that pulpal fibroblasts, as well as macrophages, may participate in the production of prostaglandin through COX-2 expression in pulpal inflammation, and might be involved in the pathogenesis of irreversible pulpitis.
Akiko Kawasaki, Kunio ISHIKAWA, Toshiyuki Suge, Hirotoshi SHIMIZU, Kazuomi SUZUKI, Takashi Matsuo and Shigeyuki EBISU : Effects of plaque control on the patency and occlusion of dentine tubules in situ., Journal of Oral Rehabilitation, Vol.28, No.5, 439-449, 2001.
(要約)
To elucidate the nature of dentine hypersensitivity, the effects of plaque control on the patency and occlusion of dentinal tubules were investigated systematically in situ using human dentine slabs embedded in partial dentures. The dentine slabs were divided into three groups. In group I, the dentine slabs were kept in an oral cavity without plaque control. In group II, plaque was removed mechanically by brushing. Plaque control was carried out chemically using chlorhexidine in group III. After being kept in the oral cavities for 1, 2 and 3 weeks, the slabs were removed from the partial denture, followed by SEM observation to determine the morphological changes of the dentinal tubules. When no attempt was made to remove plaque, the diameter of tubule orifices increased to 390% of the original values within 3 weeks. In contrast, dentinal tubules were found to be occluded, i.e. the tubule orifices became <20% of the original value within 1 week when plaque control efforts were made, using either method of plaque control. We conclude that plaque control plays one of the key roles in the patency versus occlusion of dentinal tubules, and thus in the aetiology and natural reparative process of dentine hypersensitivity.
Kinya Murakami, Katsuhiko Hirota, Takashi Matsuo and Yoichiro Miyake : Effect of antibiotics on CD15s-related antigen expressed on the Streptococcus anginosus group, Dentistry in Japan, Vol.36, 22-27, 2000.
(キーワード)
S. Anginosus Group / CD15s-Related Antigen / 抗生物質 (antibiotics)
Takeshi Kimochi, Masahiro Yoshiyama, Akihisa Urayama and Takashi Matsuo : Adhesion of a new commercial self-etching/self-priming bonding resin to human caries-infected dentin, Dental Materials Journal, Vol.18, No.4, 437-443, 1999.
(キーワード)
Resin / Adhesion / caries-infected dentin
110.
Toshiyuki Suge, Kunio ISHIKAWA, Akiko Kawasaki, Kazuomi SUZUKI, Takashi Matsuo and Shigeyuki EBISU : Evaluation of post-treatment solutions for clinical use with the calsium phosphate precipitation method, Journal of Dentistry, Vol.27, No.7, 487-496, 1999.
(要約)
We have proposed a calcium phosphate precipitation (CPP) method that occludes dentine tubules with apatitic minerals for the treatment of dentine hypersensitivity. The current CPP method uses 1 mol/l NaOH as the post-treatment solution. However, its high pH is not desirable for clinical use. The aim of this study was to evaluate several solutions, especially buffer solutions, as post-treatment solutions for the CPP method. The CPP solution was mixed with several post-treatment solutions at various mixing ratios in test tubes. The precipitates were collected, and freeze-dried in a vacuum. Their weights were measured to evaluate the feasibility of buffer solution for the post-treatment solutions. Among the solutions, we selected one buffer solution and have done further evaluation using human dentine disks. The degree of occlusion of dentine tubules was evaluated by scanning electron microscopic observation and measurement of dentine permeability. Also, the composition of the precipitate was analyzed using energy-dispersive X-ray microanalysis and powder X-ray diffraction. Among the solutions, 1 mol/l NaHCO3 gave sufficient amounts of precipitate at a relatively mild pH, and thus was considered to be a good candidate for a post-treatment solution. Scanning electron microscopic observation showed that dentine tubules were occluded with the precipitate to a depth of approximately 10 microns from the surface when the CPP treatment was done twice, and dentine permeability was reduced to 98.8%. The precipitate was dicalcium phosphate dihydrate (DCPD; CaHPO4-2H2O) when the post-treatment solution was free of NaF. In contrast, the precipitate was an apatitic mineral with CaF2 as a by-product when the post-treatment solution contained NaF. We concluded that 1 mol/l NaHCO3 containing 0.3 mol/l NaF would be suitable as the post-treatment solution for the CPP method as it has a relatively mild pH and occludes dentinal tubules well.
Fumiko Michishige, Sumiko Yoshinaga, Eriko Harada, Katsuhiko Hirota, Yoichiro Miyake, Takashi Matsuo and Susumu Yasuoka : Relationships between activity of daily living, and oral cavity care and the number of oral cavity microorganisms in patients with cerebrovascular diseases, The Journal of Medical Investigation : JMI, Vol.46, No.1-2, 79-85, 1999.
(要約)
We examined the relationships among the activity of daily living (ADL), oral cavity care, and the number of oral cavity microorganisms in 40 patients with cerebrovascular diseases (CVD). The CVD patients were classified into 4 groups, I, II, III and IV based on their ADL and the method used for oral cavity care. The ADL was highest in group I and lowest in group III. Only the patients of only group III could not eat by themselves and were receiving naso-esophageal feeding. Oral cavity care was performed by the patients themselves in groups I and IV, but was performed by caregivers in groups II and III. The group IV patients had no teeth, but could eat by themselves using full dentures. The numbers of microorganisms in the pharyngeal swabs from the 4 groups were measured and expressed as colony-forming units (cfu). The numbers of both Staphylococci spp. and Candida spp. were significantly higher in group III than in the other groups. Moreover, Pseudomonas aeruginosa was isolated only from patients of group III (in about 66%). The oral cavity care by caregivers was almost the same in groups II and III, but the numbers of oral cavity microorganisms were significantly higher in group III than in group II. These results indicated that microorganisms grow more easily in the oral cavities of CVD patients with low ADL compared with CVD patients with higher ADL, and that eating is thought to be important for the prevention of an increase of microorganisms in the oral cavity.
Hiromichi Yumoto, Hideaki Nakae, Keiko Fujinaka, Shigeyuki Ebisu and Takashi Matsuo : Interleukin-6 (IL-6) and IL-8 are induced in human oral epithelial cells in response to exposure to periodontopathic Eikenella corrodens, Infection and Immunity, Vol.67, No.1, 384-394, 1999.
Masahiro Yoshiyama, Takashi Matsuo, Shigeyuki Ebisu and David Pashley : Regional bond strengths of self-etching/self-priming adhesive systems, Journal of Dentistry, Vol.26, No.7, 609-616, 1998.
(要約)
The purpose of this study was to measure the regional tensile bond strengths (TBS) at various portions of human tooth enamel and dentin, and to observe the resin-dentin interfaces of two commercially available self-etching/self-priming adhesive systems by scanning electron microscopy (SEM). Twelve extracted human cuspid teeth were used to measure TBS and four additional teeth were used for scanning electron microscopy (SEM). Outer enamel and dentin were removed from the labial tooth surfaces to form a long cavity preparation into middle dentin extending from the mid-crown to the apex of the root within the same tooth. Either Clearfil Liner Bond 2 (LB 2, Kuraray) or Fluoro Bond (FB, Shofu) was bonded to the surfaces, and covered with resin-composite. The resin-bonded teeth were serially sliced at right angles to the long axis of the tooth, and the bonded surfaces were trimmed to give a bonded cross-sectional surface area of 1 mm2 for TBS tests. LB 2 and FB showed significantly higher TBS in coronal, cervical and middle root dentin than that in enamel and apical root dentin. SEM showed that the thickness of the hybrid layer of both systems was about 1.0 microm in coronal, cervical and middle root dentin, and less than 0.5 microm in apical root dentin. These results suggested that the self-etching/self-priming systems produce good adhesion in coronal, cervical and middle root dentin by creating thin hybrid and transitional layers, but bonding to enamel and apical root dentin should be improved.
Katsuhiko Hirota, Kinya Murakami, Ken Nemoto, Tsuneko Ono, Takashi Matsuo, Hiromi Kumon and Yoichiro Miyake : Fosfomycin reduces CD15s-related antigen expression of Streptococcus pyogenes, Antimicrobial Agents and Chemotherapy, Vol.42, No.5, 1083-1987, 1998.
(要約)
We have previously shown the immunological mimicry of human sialyl-Lewis(x) (CD15s) by a surface antigen of Streptococcus pyogenes. This mimicking surface antigen may act as a ligand to the selectin family and may induce antibody production against CD15s on host cells, suggesting a possible role in the pathogenesis of S. pyogenes. In this study, the effects of antibiotics on the CD15s-related antigen expression of S. pyogenes were examined at a concentration below the MIC (sub-MIC). The amounts of CD15s on the surfaces of S. pyogenes cells and on the surfaces of S. pyogenes biofilms were determined by a whole-cell enzyme-linked immunosorbent assay and by laser scanning fluorescence microscopy, respectively, by using an anti-CD15s monoclonal antibody. At the sub-MICs, fosfomycin (1R,2S-1,2-epoxypropyl phosphonic acid), its enantiomer (1S,2R-1,2-epoxypropyl phosphonic acid), and benzylpenicillin significantly inhibited the CD15s expression of all strains studied. The effects of fosfomycin and its enantiomer on biofilms were also observed by scanning electron microscopy. Incubation of S. pyogenes with the sub-MIC of fosfomycin or its enantiomer, which has no antibacterial activity, reduced the amount of CD15s on the biofilm surface and made it smooth. These results suggest that fosfomycin or its enantiomer might be useful for preventing S. pyogenes adherence to human CD15s receptors and the resulting immunological pathogenicity.
Yuichiro Noiri, Kazumi Ozaki, Hideaki Nakae, Takashi Matsuo and Shigeyuki Ebisu : An immunohistochemical study on the localization of Porphyromonas gingivalis, Campylobacter rectus and Actinomyces viscosus in human periodontal pockets, Journal of Periodontal Research, Vol.32, No.7, 598-607, 1997.
Hiromichi Yumoto, Hiroyuki Azakami, Hideaki Nakae, Takashi Matsuo and Shigeyuki Ebisu : Cloning, sequencing and expression of an Eikenella corrodens gene encoding a component protein of the lectin-like adhesin complex, Gene, Vol.183, No.1-2, 115-121, 1996.
123.
Hiroyuki Azakami, Hiromichi Yumoto, Hideaki Nakae, Takashi Matsuo and Shigeyuki Ebisu : Molecular analysis of the gene encoding a protein component of the Eikenella corrodens adhesin complex that is close to the carbohydrate recognition domain, Gene, Vol.180, No.1-2, 207-212, 1996.
124.
Takashi Matsuo, Tadashi Nakanishi, Hirotoshi Shimizu and Shigeyuki Ebisu : A clinical study of direct pulp capping applied to carious-exposed pulps, Journal of Endodontics, Vol.22, No.10, 551-556, 1996.
125.
Takashi Matsuo, Chisato Tagawa, Kazumi Ozaki, Yuichiro Noiri and Shigeyuki Ebisu : A Histological Analysis of T and B Cells in Periodontal Tissue Destruction, Oral Medicine & Pathology, Vol.1, No.2, 77-83, 1996.
(要約)
We immunohistologically examined the cell densities and distribution of T and B cells in periodontitis and analyzed their relationship in terms of periodontal tissue destruction. Specimens were labeled with monoclonal antibodies for T cells (CD3) and B cells (CD20) and developed using the double staining technique. CD3^+ cells appeared in low density infiltrates and increased gradually with the number of total infiltrates; in contrast, CD20^+ cells did not appear in small infiltrates but increased rapidly with the number of infiltrates. The number of fibroblastic cells negatively correlated with that of the total infiltrates and CD20^+ cells, but it did not correlate with the number of CD3^+ cells. These findings suggest that the T cells infiltrated into the inflamed sites at the onset and increased gradually with the development of inflammation and that B cells infiltrated later than T cells and their number increased rapidly with inflammation.
Takashi Matsuo, Tadashi Nakanishi and Shigeyuki Ebisu : Immunoglobulins in periapical exudates of infected root canals: correlations with the clinical findings of the involved teeth, Endodontics & dental traumatology, Vol.11, No.2, 95-99, 1995.
128.
Tadashi Nakanishi, Takashi Matsuo and Shigeyuki Ebisu : Quantitative analysis of immunoglobulins and inflammatory factors in human pulpal blood from exposed pulps, Journal of Endodontics, Vol.21, No.3, 131-136, 1995.
Hideaki Nakae, Hiromichi Yumoto, Takashi Matsuo and Shigeyuki Ebisu : Mitogenic stimulation of murine B lymphocytes by the N-acetyl-D-galactosamine specific bacterial lectin-like substance from Eikenella corrodens, FEMS microbiology letters, Vol.116, No.3, 349-353, 1994.
P C Quan, S Watanabe, F Vuillier, R Pires, Takashi Matsuo, M Stanislawski, J Pillot and P J Bouvet : Purification and partial amino acid sequence of suppressive lymphokine from a CD8+ CD57+ human T hybridoma, Immunology, Vol.78, No.2, 205-209, 1993.
139.
Takashi Matsuo, Shigeyuki Ebisu, Yoshio Shimabukuro, Tsuyoshi Ohtake and Hiroshi Okada : Quantitative analysis of immunocompetent cells in human periapical lesions: correlations with clinical findings of the involved teeth, Journal of Endodontics, Vol.18, No.10, 497-500, 1992.
H Okada, Y Shimabukuro, Y Kassai, Hiro-O Ito, Takashi Matsuo, S Ebisu and Y Harada : The Function of Gingival Lymphocytes on the Establishment of Human Periodontitis, Advances in Dental Research, Vol.2, No.2, 364-367, 1988.
147.
Takashi Matsuo, Shinichiro Watanabe, Bouvet Jean-Pierre, Kolb Jean-Pierre and P Canh Quan : Suppressor factor secreted by T hybridoma established from peripheral blood lymphocytes of a bone marrow transplantation patient. I. Establishment of human T-cell hybridoma and partial characterization of suppressor factor, Cellular Immunology, Vol.116, No.2, 450-466, 1988.
(要約)
A stable human T-cell hybridoma was established by cell fusion between activated human peripheral blood lymphocytes from an allogeneic bone marrow transplantation patient and the JD1-17 cell line, a subclone of the human T leukemia Jurkat cell line. This hybrid clone 1-8, which bore the surface phenotype of suppressor cells (CD8+HNK1+), spontaneously secreted a factor which, at high dilutions, suppressed the responses of T and B cells induced by mitogens and alloantigens. This suppressor factor was found to be heat-resistant (56 degrees C, 30 min), stable at alkaline but not acid pH, unaffected by 2-mercaptoethanol, and sensitive to trypsin. Preparative isoelectric focusing revealed an isoelectric point of 5.35. The suppressor activity was selectively absorbed by blast T cells. By gel filtration on Sephacryl S-200 and HPLC, the suppressor activity was found in two peaks corresponding to 40-45 kDa (monomer) and 90-95 kDa (dimer).
(キーワード)
Absorption / Biological Factors / Bone Marrow Transplantation / Carbohydrates / Cell Cycle / Chromatography, Gel / Chromatography, High Pressure Liquid / Hot Temperature / Humans / Hybridomas / Hydrogen-Ion Concentration / Immune Tolerance / Interferons / Interleukin-2 / Interleukin-4 / Interleukins / Lymphocyte Activation / Phenotype / T-Lymphocytes, Regulatory
Hiro-O Ito, Yasushi Harada, Takashi Matsuo, Shigeyuki Ebisu and Hiroshi Okada : Possible role of T cells in the establishment of IgG plasma cell-rich periodontal lesion--Augmentation of IgG synthesis in the polyclonal B cell activation response by autoreactive T cells, Journal of Periodontal Research, Vol.23, No.1, 39-45, 1988.
松尾 敬志, 中西 正, 湯本 浩通, 平尾 功治, 篠原 千尋, Toshihiko Tominaga : 感染根管治療の考え方, Journal of Oral Health and Biosciences, Vol.28, No.2, 87-92, 2016年.
(要約)
It is a well-established fact that bacterial infection of the dental pulp ultimately results in the formation of apical periodontitis. And apical periodontitis may develop periapical leisons consisting granulomas and cysts. In this article, we show the pathologic condition of infected root canal systems and periapical lesions immunohistologically. We also describe the possible causes of refractory apical periodontitis that conventional root canal therapy could not work effectively. Finally,we explain our newly developing treatment for refractory apical periodontitis named electro-magnetic apical treatment, and illustrate clinical efficacy of this treatment.
Toshiyuki Suge and Takashi Matsuo : Effects of ammonium hexafluorosilicate concentration on crystallinity of hydroxyapatite powder and enamel, Key Engineering Materials, Vol.529-530, 526-530, 2013.
The sbjects were classified 3 groups. Oral care was effectively carried out at 10o'clok in the morning group, 15o'clok in the midday group and 19o'clok in the night group. The number of Streptococci spp. and Staphlococci spp. in the samples was quantitatively measured. These results showed that effect of mouth care is not influenced by the time when it is done, as far as it is effectively done once in a day, and suggested that in the subjects who can not perform oral care by themselves, and can not receive oral care service several times in a day. It is important to perform effective mouth care which remove completely dental plaque, once in a day, by caregivers.
Nur Asikin, Katsuhiko Hirota, Hiromichi Yumoto, Kouji Hirao, Kanako Takahashi, Keiji Murakami, Takashi Matsuo and Yoichiro Miyake : Biological Roles of Extracellular DNA in Bacterial Infection, Asean Plus and Tokushima Joint International Conference, Program and Proceeding Book, 47-48, 2012.
3.
Katsuhiko Hirota, Takeyoshi Yoneyama, Mayumi Sakamoto, Hiroshi Miyamoto, Masanori Kurihara, Shizuo Kayama, Keiji Murakami, Hiromichi Yumoto, Takashi Matsuo and Yoichiro Miyake : High prevalence of Pseudomonas aeruginosa from oropharyngeal biofilm in patients with cerebrovascular infarction and dysphagia., Chest, Vol.138, No.1, 237-238, 2010.
Hiromichi Yumoto, Kouji Hirao, Yuki Hosokawa, Hitomi Kuramoto, Daisuke Takegawa, Tadashi Nakanishi and Takashi Matsuo : The Roles of Odontoblasts in Dental Pulp Innate Immunity, Japanese Dental Science Review, Vol.54, No.3, 105-117, Aug. 2018.
(要約)
Odontoblasts located in the outermost layer of dental pulp form a natural barrier between mineralized tissues, dentin, and soft tissues, dental pulp, of the vital tooth, and they first recognize caries-related pathogens and sense external irritations. Therefore, odontoblasts possess a specialized innate immune system to fight oral pathogens invading into dentin. Generally, the rapid initial sensing of microbial pathogens, especially pathogen-associated molecular patterns (PAMPs) shared by microorganisms, are mediated by pattern recognition receptors (PRRs), such as Toll-like receptor and the nucleotide-binding oligomerization domain (NOD). The innate immune responses in odontoblasts initiated by sensing oral pathogens provide host protective events, such as inflammatory reactions, to produce a variety of pro-inflammatory mediators, including chemokines and cytokines. These attract various inflammatory cells and cause antibacterial reactions, such as the production of defensins, to kill microorganisms in the proximal region of the odontoblast layer. This review focuses on innate immunity, especially cellular and molecular mechanisms regarding the sensing of PAMPs from oral pathogens by PRRs, in odontoblasts and provides information for future studies for the development of novel therapeutic strategies, including diagnosis and treatment, to prevent exceeding dental pulp inflammation and preserve the dental pulp tissues.
Hiromichi Yumoto, Kouji Hirao, Yuki Hosokawa, Hitomi Kuramoto, Katsuhiko Hirota, Yoichiro Miyake and Takashi Matsuo : Streptococcal histone-like DNA binding protein induces MINCLE expression on monocytes, 95th General Session & Exhibition of the International Association for Dental Research, San Francisco, Mar. 2017.
2.
Yuki Hosokawa, Hiromichi Yumoto, Kouji Hirao, Hitomi Kuramoto, Tadashi Nakanishi, Daisuke Takegawa and Takashi Matsuo : Anti-inflammatory effects of polyphenols on rat odontoblastic cells, 95th General Session & Exhibition of the International Association for Dental Research (San Francisco), Mar. 2017.
3.
Daisuke Takegawa, Tadashi Nakanishi, Kouji Hirao, Hiromichi Yumoto, Yuki Hosokawa and Takashi Matsuo : Interferon-gamma modulates the innate immune response in odontoblast-like cells, 95th General session and exhibition of the IADR, Mar. 2017.
4.
Tadashi Nakanishi, Daisuke Takegawa, Kouji Hirao, Hiromichi Yumoto, Yuki Hosokawa and Takashi Matsuo : Effect of interleukin-17A on CCL20 production from odontoblast-like cells, 95th General session and exhibition of the IADR, Mar. 2017.
5.
Susilowati Heni, Keiji Murakami, Takashi Amoh, Katsuhiko Hirota, Yoichiro Miyake, Hiromichi Yumoto and Takashi Matsuo : Royal Jelly inhibits Pseudomonas aeruginosa adherence and reduces excessive inflammatory resoponses in human epithelial cells, 2016 IADR, Seoul, Jun. 2016.
6.
Toshihiko Tominaga, Kazuki Takahira, Hiromichi Yumoto, Takashi Matsuo, Naoki Bando and Kouji Hirao : EMAT (Electromagnetic Apical Treatment): Clinical Application of Pulsed Electric Current Energization for Root Canal Treatment, 2015 Annual Session of the American Association of Endodontists, Seattle (USA), May 2015.
7.
Tadashi Nakanishi, Daisuke Takegawa, Hiromichi Yumoto, Kouji Hirao and Takashi Matsuo : Interleukin-17 Regulates Inflammatory Mediator Expression in IL-1beta-stimulated Dental Pulp Cells, 93th General Session & Exhibition of the International Association for Dental Research, Boston (USA), Mar. 2015.
8.
Shindo Satoru, Yoshitaka Hosokawa, Ikuko Hosokawa, Kazumi Ozaki and Takashi Matsuo : The effect of genipin on dendritic cells activation, 93rd General Session & Exhibition of the IADR, Boston, Mar. 2015.
9.
Ikuko Hosokawa, Yoshitaka Hosokawa, Shindo Satoru, Kazumi Ozaki and Takashi Matsuo : Melatonin inhibits inflammatory mediators productions in human periodontal ligament cells, 93rd General Session & Exhibition of the IADR, Boston, Mar. 2015.
10.
Yoshitaka Hosokawa, Ikuko Hosokawa, Satoru Shindo, Kazumi Ozaki and Takashi Matsuo : IL-4 modulates chemokine productions from IL-1-stimulated human periodontal ligament cells, 93rd General Session & Exhibition of the IADR, Boston, Mar. 2015.
11.
Kouji Hirao, Hiromichi Yumoto, Chihiro Shinohara, Hosokawa Yuki and Takashi Matsuo : Functional roles of NOD1 in odontoblasts on pulp innate immunity, The 62nd annual meeting of japanese association for dental research, 99, Dec. 2014.
12.
Hiromichi Yumoto, Takashi Matsuo, Katsuhiko Hirota, Kouji Hirao, Miyazaki Tsuyoshi, Keiji Murakami, Natsumi Fujiwara and Yoichiro Miyake : Anti-inflammatory and protective effects of 2-methacryloyloxyethyl-phosphorylcholine-polymer on oral epithelial cells, 92th General Session & Exhibition of the International Association for Dental Research (Cape Town, South Africa), Jun. 2014.
13.
Satoru Shindo, Yoshitaka Hosokawa, Ikuko Hosokawa and Takashi Matsuo : The effect of genipin on MMP-1 and MMP-3 expression in IL-1beta-stimulated human periodontal ligament cells, The 15th Joint Scientific Meeting of JSCD-KACD, Gyeongju, Korea, Nov. 2013.
14.
Hiromichi Yumoto, Katsuhiko Hirota, Kouji Hirao, Keiji Murakami, Yoichiro Miyake and Takashi Matsuo : Pathogenic roles of Streptococcal histone-like protein in microbial infection, The 15th Joint Scientific Meeting of JSCD-KACD (Gyeongju, Korea), Nov. 2013.
15.
Toshihiko Tominaga, Naoki Bando, Hiromichi Yumoto and Takashi Matsuo : Trial of Electro-Magnetic Wave Irradiation Therapy for Vertical Root Fracture as Tooth Conservative Therapy: a Case report, The 9th World Endodontic Congress, Tokyo, Japan, May 2013.
16.
Naoki Bando, Toshihiko Tominaga, Hiromichi Yumoto and Takashi Matsuo : Clinical Application of Electro-Magnetic Wave Irradiation to the Treatment for Periapical Periodontitis; Case Report, The 9th World Endodontic Congress, Tokyo, Japan, May 2013.
17.
Tadashi Nakanishi, Daisuke Takegawa, Kouji Hirao, Hiromichi Yumoto, Kanako Takahashi and Takashi Matsuo : Prostaglandin F2a Regulates Cytokine Expression in Dental Pulp Cells, 91th General Session & Exhibition of the IADR, Seattle, USA, Mar. 2013.
18.
Daisuke Takegawa, Tadashi Nakanishi, Kouji Hirao, Hiromichi Yumoto, Kanako Takahashi and Takashi Matsuo : Interferon-g modulates the innate immune response of dental pulp cells, 91th General Session & Exhibition of the IADR, Seattle, USA, Mar. 2013.
19.
Nur Asikin, Katsuhiko Hirota, Hiromichi Yumoto, Kouji Hirao, Kanako Takahashi, Keiji Murakami, Takashi Matsuo and Yoichiro Miyake : Biological Roles of Extracellular DNA in Bacterial Infection, Asean Plus and Tokushima Joint International Conference, Yogyakarta, Dec. 2012.
20.
Hiromichi Yumoto, Toshihiko Tominaga, Kouji Hirao, Naoki Bando, Takashi Sumitomo, Kanako Takahashi and Takashi Matsuo : Signaling Pathways and Gene Expression Activated by Electro-Magnetic-Wave in Osteoblastic-Cells, 90th General Session and Exhibition of the IADR, Iguazu Falls, Brazil, Jun. 2012.
21.
Kouji Hirao, Hiromichi Yumoto, Katsuhiko Hirota, Dali Liu, Nur Asikin, Kanako Takahashi, Takashi Matsuo and Yoichiro Miyake : Global Gene Analysis in Monocytes Stimulated with Streptococcal Histone-Like Protein, 90th General Session and Exhibition of the IADR, Iguazu Falls, Brazil, Jun. 2012.
22.
Toshihiko Tominaga, Hiromichi Yumoto and Takashi Matsuo : Clinical Application of Electro-Magnetic Wave Irradiation for Infected Root Canal Treatment -EMAT (Electro-Magnetic Apical Treatment) -, 2012 Annual Session of the American Association of Endodontists, Boston, USA, Apr. 2012.
23.
Hiromichi Yumoto, Toshihiko Tominaga and Takashi Matsuo : Electro-Magnetic Wave Irradiation Stimulates Osteoblasts to Enhance Cell Growth and Gene Expression of Growth Factors, 2012 Annual Session of the American Association of Endodontists, Boston, USA, Apr. 2012.
24.
Katsuhiko Hirota, Hiromichi Yumoto, Nur Asikin, Kouji Hirao, Kanako Takahashi, Liu Dali, Keiji Murakami, Takashi Matsuo and Yoichiro Miyake : Gene Expression Profiling of Human Monocytic THP-1 Cells Stimulated With Extracellular Streptococcus intermedius Histone-Like DNA Binding Protein, International Union of Microbiological Societies 2011 Congress, Sep. 2011.
25.
Takashi Sumitomo, Toshihiko Sumitomo, Naoki Bando, Saki Hirao, Shinya Kako, Hiromichi Yumoto and Takashi Matsuo : A Case Report; Electro-Magnetic Apical Treatment, The 9th KAE-JEA Joint Meeting, Jul. 2011.
26.
Yoichiro Miyake, Katsuhiko Hirota, Hiromichi Yumoto, N Yamamoto, K Miyamoto and Takashi Matsuo : Inhibition of the adherence of oral bacterial by MPC-polymer, EUROBIOFILMS 2011 -Second European Congress on Microbial Biofilms-Basic and Dlinical Aspedts-, Copenhagen, Jul. 2011.
27.
Tsuyoshi Fujita, Hiromichi Yumoto, Kazuhisa Ouhara, Hideki Shiba, Tsuyoshi Miyagawa, S Matsuda, Hiroyuki Kawaguchi, Takashi Matsuo and Hidemi Kurihara : Irsogladine Maleate Regulates Gingival Epithelial Barrier Functions In Inflammatory Condition, 89th General Session & Exhibition of the International Association for Dental Research, Mar. 2011.
28.
Tadashi Nakanishi, Kouji Hirao, Takegawa Daisuke, Hiromichi Yumoto, Kanako Takahashi and Takashi Matsuo : Catechins suppress cyclooxygenase-2 expression in human dental pulp cells, 88th General Session and Exhibition of the IADR, Jul. 2010.
29.
Toshihiko Tominaga, Hiromichi Yumoto and Takashi Matsuo : Novel Therapeutic Approach of Electro-Magnetic Wave for Infected Root Canal, 88th General Session and Exhibition of the IADR, Jul. 2010.
30.
Takegawa Daisuke, Tadashi Nakanishi, Kouji Hirao, Hiromichi Yumoto, Kanako Takahashi and Takashi Matsuo : Interferon-gamma enhances toll-like receptor ligand-induced cytokine production in pulpal cells, 88th General Session and Exhibition of the IADR, Jul. 2010.
31.
Hiromichi Yumoto, T. TOMINAGA, Kouji Hirao, Tomoko Kimura and Takashi Matsuo : Inactivation and Bactericidal Activity against Oral Bacteria of Electro-Magnetic Wave, 88th General Session and Exhibition of the IADR, Jul. 2010.
32.
Toshihiko Tominaga, Takashi Sumitomo, Hiromichi Yumoto and Takashi Matsuo : The effects of Electro-Magnetic-Wave on bacterial viability and osteoblastic cell proliferation, American Association of Endodontics, 2009 Annual Session, Apr. 2009.
33.
Ikuko Hosokawa, Yoshitaka Hosokawa, Kazumi Ozaki, Hiromichi Yumoto, Hideaki Nakae and Takashi Matsuo : Pro-inflammatory Roles of NOD2 in Human Gingival Fibroblasts, The 3rd International Symposium on "Oral Sciences to Improve the Quality of Life", Tokushima, Sep. 2008.
34.
Hiromichi Yumoto, Liu Dali, Keiji Murakami, Katsuhiko Hirota, Hideaki Nagamune, Shizuo Kayama, Takashi Matsuo and Yoichiro Miyake : Gene-regulatory and Physiological Functions of Histone-like Protein from Streptococci, 86th General Session and Exhibition of the International Association for Dental Research, Toronto (Canada), Jul. 2008.
35.
Ikuko Hosokawa, Yoshitaka Hosokawa, Kazumi Ozaki, Hiromichi Yumoto, Hideaki Nakae and Takashi Matsuo : Pro-inflammatory Roles of NOD2 in Human Gingival Fibroblasts, 86th General Session and Exhibition of the International Association for Dental Research, Toronto, Jul. 2008.
36.
Kayo Mukai, Tadashi Nakanishi, Hiromichi Yumoto, Kanako Takahashi, Kouji Hirao and Takashi Matsuo : Effects of Catechins on Cytokine Production in Human Odontoblast-like Cells, 86th General Session and Exhibition of the International Association for Dental Research, Jul. 2008.
37.
Kouji Hirao, Hiromichi Yumoto, Tadashi Nakanishi, Kanako Takahashi, Kayo Mukai, Miho Yoneda and Takashi Matsuo : Functional Analysis of Pattern Recognition Receptor Expression in Pulpal Cells, 86th General Session and Exhibition of the International Association for Dental Research, Jul. 2008.
38.
Yoshitaka Hosokawa, Ikuko Hosokawa, Kazumi Ozaki, Hideaki Nakae and Takashi Matsuo : Th17 cytokines enhance CCL20 production by human gingival fibroblasts., 86th General Session and Exhibition of the International Association for Dental Research, Toronto, Jun. 2008.
39.
Liu Dali, Hiromichi Yumoto, Keiji Murakami, Katsuhiko Hirota, Shizuo Kayama, Takashi Matsuo and Yoichiro Miyake : The essentiality and involvements of streptocuccus intermedius histone-like protein in bacterial viability and normal growth, The 2nd International Symposium on "The Future Direction of Oral Sciences in the 21st Century", Tokushima, Dec. 2007.
40.
Tomoko Kimura, Kazumi Ozaki, Keiji Murakami, Kikumi Fujisaka and Takashi Matsuo : The Analysis of role of Phosphoenolpyruvate-Dependent Sucrose Phosphotransferase System in pathogenicity of Streptococcus mutans., 第1回国際シンポジウム・FDワークショップ, Awaji, Mar. 2007.
41.
Kouji Hirao, Hiromichi Yumoto, Kanako Takahashi, Kayo Mukai and Takashi Matsuo : Expression and Functional Analyses of Pattern recognition Receptor (PRR) in Cultured Human Dental Pulp Cells, The 1st international symposium and workshop "The Future Direction of Oral Sciences in the 21st century", Mar. 2007.
42.
Kouji Hirao, Hiromichi Yumoto, Kanako Takahashi, Kayo Mukai and Takashi Matsuo : Expression and Functional Analyses of Pattern recognition Receptor (PRR) in Cultured Human Dental Pulp Cells, The 8th Joint-Scientific Meeting between KACD & JSCD, Busan(Korea), Nov. 2006.
43.
Kayo Mukai, Tadashi Nakanishi, Yoshitaka Hosokawa, Kanako Takahashi and Takashi Matsuo : Inhibitory effects of catechins on inflammatory reactions in pulp fibroblasts, 84th General Session & Exhibition of the IADR, Brisbane, Jun. 2006.
44.
Miho Yoneda, Hideaki Nakae, Hiromichi Yumoto, Chihiro Shinohara and Takashi Matsuo : Expression pattern of claudin family in human gingival tissues, 84th General Session and Exhibition of the International Association for Dental Research, Jun. 2006.
45.
Akiko Kawasaki, Toshiyuki Suge, Kunio ISHIKAWA and Takashi Matsuo : Effect of ammonium hexafluorosilicate on acid resistance of enamel and dentin, Sydney, May 2004.
46.
Toshiyuki Suge, Akiko Kawasaki, Kunio Ishikawa and Takashi Matsuo : Occlusion of dentinal tubules with ammonium hexafluorosilicate for treatment of dentin hypersensitivity, 7th World Biomaterials Congress, Sydney, May 2004.
47.
Natsuko Ida, Kazumi Ozaki, Shinji Suenobu, Kanako Takahashi, Daisuke Yamaguchi and Takashi Matsuo : Opsonized Streptococcus mutans influences maturation of human dendritic cells, The 82nd General Session of IADR Meeting, Hawaii, Mar. 2004.
48.
Kazumi Ozaki and Takashi Matsuo : Expression of adrenomedullin and its receptors in inflamed gingival tissue, The 82nd General Session of IADR Meeting, Hawaii, Mar. 2004.
49.
Tadashi Nakanishi, Tomohiko Adachi, Kanako Takahashi and Takashi Matsuo : Expression of interferon-gamma -inducible protein 10/CXCL10 in inflamed dental pulp, 82th General Session of the IADR(INternational Association of Dental Research), Honolulu, Mar. 2004.
50.
Kanako Takahashi, Tadashi Nakanishi, Tomohiko Adachi, Natsuko Ida, Hideaki Nakae and Takashi Matsuo : Effects of Streptococcus mutans on MIP-3 α production by macrophages, 82th General Session of the IADR(INternational Association of Dental Research), Honolulu, Mar. 2004.
51.
Natsuko Ida, Kazumi Ozaki, Shinji Suenobu, Kanako Takahashi, Daisuke Yamaguchi and Takashi Matsuo : Opsonized Streptococcus mutans influences maturation of human dendritic cells., Seoul, Nov. 2003.
52.
Takuya Shibutani, Kazumi Ozaki and Takashi Matsuo : Comparison of root canal preparation by three Ni-Ti instruments, Seoul, Nov. 2003.
53.
Daisuke Yamaguchi, Tadashi Nakanishi, Kanako Takahashi, Hideaki Nakae and Takashi Matsuo : Expression of macrophage inflammatory protein-3alpha/CCL20 in human gingival epithelial cells, 81th General Session of the IADR(INternational Association of Dental Research), エルサレム, Jun. 2003.
54.
Kazumi Ozaki, Shinji Suenobu, Natsuko Ida and Takashi Matsuo : Bacterial Viability Assay at an Electron Microscopic Leved, The 81th General Session of IADR Meeting, Goteborg, Sweden, Jun. 2003.
55.
Shinji Suenobu, Kazumi Ozaki and Takashi Matsuo : The Analysis of mRNA Expression of Glucosyltransferases and Dextranase A from Streptococcs mutans Adhered to the Dentin, The 81th General Session of IADR Meeting, Goteborg, Sweden, Jun. 2003.
56.
Takuya Shibutani, Kazumi Ozaki and Takashi Matsuo : Comparison of the Effects of Three Ni-Ti Preparation Techniques by Micro-computed Tomography, The 81th General Session of IADR Meeting, Goteborg, Sweden, Jun. 2003.
57.
Akiko Kawasaki, Toshiyuki Suge, Kunio Ishikawa, Kazumi Ozaki, Takashi Matsuo and Shigeyuki Ebisu : Ammonium hexafluorosilicate increase acid resistance of bovine enamel and dentin, Journal of Dental Research, Vol.82, No.Special Issue B, B-71, Goteborg, Jun. 2003.
58.
Toshiyuki Suge, Kunio Ishikawa, Akiko Kawasaki, Takashi Matsuo and Shigeyuki Ebisu : Duraiton of ammonium hexafluorosilicate for dentinal tubules occlusion, Journal of Dental Research, Vol.82, No.Special Issue B, B-47, Goteborg, Jun. 2003.
59.
Hiromichi Yumoto, Katsuhiko Hirota, Hideaki Nagamune, Kazuto Ohkura, Keiji Murakami, Masayoshi Yamada, Hideaki Nakae, Yoichiro Miyake and Takashi Matsuo : Molecular analysis of histone-like protein from Streptococcus intermedius, 80th General Session and Exhibition of the International Association for Dental Research, San Diego(USA), Mar. 2002.
60.
Masayoshi Yamada, Hiromichi Yumoto, Chihiro Shinohara, Hideaki Nakae, Shigeyuki Ebisu and Takashi Matsuo : Eikenella corrodens Induces Endothelial Cells via MAPK Family pathways, 80th General Session and Exhibition of the International Association for Dental Research, San Diego(USA), Mar. 2002.
61.
Yoshitaka Hosokawa, Tadashi Nakanishi, Daisuke Yamaguchi, Hiromichi Yumoto and Takashi Matsuo : LARC-CCR6 Interactions play an important role in CD4+ T cell accumulation in periodontal diseased tissue, 80th General Session and Exhibition of the International Association for Dental Research, San Diego(USA), Mar. 2002.
62.
Tadashi Nakanishi, Kanako Takahashi, Yoshitaka Hosokawa, Daisuke Yamaguchi, Chihiro Shinohara, Hiromichi Yumoto and Takashi Matsuo : Expression of macrophage inflammatory protein-3 α in human pulpitis, 80th General SEssion ofthe IADR(International Association of Dental Research), San Diego(USA), Mar. 2002.
63.
Akiko Kawasaki, Toshiyuki Suge, Kunio ISHIKAWA, Kazumi Ozaki and Takashi Matsuo : Ammonium hexafluorosilicate increased acid resistance of bovine enamel and dentin, Transactions of the fourth international congress on dental materials, Vol.16, 195, Honolulu, Jan. 2002.
64.
Toshiyuki Suge, Kunio ISHIKAWA, Akiko Kawasaki, Takashi Matsuo and Shigeyuki EBISU : Effects of ammonium hexafluorosilicate on dental tubules' occlusion, Transactions of the fourth international congress on dental materials, Vol.16, 253, Honolulu, Jan. 2002.
65.
Takashi Matsuo, Keiko Fujinaka, Yoshiko Yoshida and Kazumi Ozaki : Scanning electron microscope observations in the apices of roots with refractory apical periodontitis, The 3rd Joint Scientific Meeting between KACD and JSCD, Seoul, Nov. 2001.
66.
Tadashi Nakanishi, Kanako Takahashi, Kazumi Ozaki, Hideaki Nakae and Takashi Matsuo : An immunohistological study on the localization of selected bacteria and chemokines in human deep-carious teeth, The International Conference on Dentin/Pulp complexs, Chiba, Jul. 2001.
67.
Akiko Kawasaki, Toshiyuki Suge, Kunio ISHIKAWA, Kazuomi SUZUKI, Takashi Matsuo and Shigeyuki EBISU : Effects of ammonium hexafluorosilicate on occluion of dentinal tubules, Journal of Dental Research, Vol.80, No.Special Issue, 750, Makuhari, Jun. 2001.
Hideaki Nakae, Masayoshi Yamada, Hiromichi Yumoto, Chihiro Shinohara and Takashi Matsuo : Bacterial lectin of Eikenella corrodens induces cytokines and adhesion molecules, Modern Periodontology New Directions in 21st Century-, Kanagawa(Japan), Jun. 2001.
70.
Hiromichi Yumoto, Daisuke Yamaguchi, Yoshitaka Hosokawa, Chihiro Shinohara, Tadashi Nakanishi, Hirotoshi Shimizu, Masayoshi Yamada, Hideaki Nakae and Takashi Matsuo : Expression of IL-18 and IL-18 receptor in human periodontally diseased tissue, Modern Periodontology New Directions in 21st Century-, Kanagawa(Japan), Jun. 2001.
71.
Chihiro Shinohara, Hiromichi Yumoto, Masayoshi Yamada, Hideaki Nakae, Shigeyuki Ebisu and Takashi Matsuo : Secreted Products from Eikenella corrodens Activate Endothelial cells., The 86th Annual Meeting of the American Academy of Periodontology inconjunction with the Japanese Society of Periodontology, Hawaii(USA), Sep. 2000.
72.
Masayoshi Yamada, Hiromichi Yumoto, Hideaki Nakae, Chihiro Shinohara, Ebisu Shigeyuki and Takashi Matsuo : Activation of mitogen-activated protein kinases in oral epithelial cells by Secreted Eikenella corrodens, The 86th Annual Meeting of the American Academy of Periodontology inconjunction with the Japanese Society of Periodontology, Hawaii(USA), Sep. 2000.
73.
Hiromichi Yumoto, Hideaki Nakae, Masayoshi Yamada, Shigeyuki Ebisu and Takashi Matsuo : Eikenella corrodens induces prostaglandin E2 production and cyclooxygenase-2 expression in oral epithelial cells, 11th International Conference on Periodontal Research, Goteborg(Sweden), Jun. 1999.
74.
Hideaki Nakae, Masayoshi Yamada, Hiromichi Yumoto, Shigeyuki Ebisu and Takashi Matsuo : Infection of human oral epithelial cells with Eikenella corrodens upregulates ICAM-1 expression, 11th International Conference on Periodontal Research, Goteborg(Sweden), Jun. 1999.
75.
Kazumi Ozaki and Takashi Matsuo : A three-dimensional examination of morphological changes in experimental cementum caries lesions, 77th General Session of IADR Meeting, Vancouver, Mar. 1999.
76.
Akiko Kawasaki, ISHIKAWA Kunio, Toshiyuki Suge, SUZUKI Kazuomi, Takashi Matsuo and EBISU Shigeyuki : In vivo evaluation of plaque control on dentin hypersensitivity, Journal of Dental Research, Vol.77, No.Special Issue, 571, Nice, Jun. 1998.
77.
Toshiyuki Suge, ISHIKAWA Kunio, Akiko Kawasaki, SUZUKI Kazuomi, Takashi Matsuo and EBISU Shigeyuki : Comparison of dentinal tubules occlusion-CPP, Resin and Potassium Oxalate, Journal of Dental Research, Vol.77, No.Special Issue, 651, Nice, Jun. 1998.
78.
Masahiro Yoshiyama and Takashi Matsuo : Adhesion to Wedge-shaped Defects and Treatments of Dentin Hypersensitivity, Modern Trends in Adhesive Dentistry Proceedings of the Adhesive Dentistry Forum `98 in Sapporo, 28-37, Sapporo, Feb. 1998.
79.
Akihisa Urayama, Takao Hanawa, Kenzo Asaoka and Takashi Matsuo : Bacterial adhesion to saliva-coated titanium surface, Transactions of the 3rd International Congress on Dental Materials, 244, Honolulu, Nov. 1997.
80.
Chihiro Shinohara, Kikuji Yamashita, Seiichiro Kitamura and Takashi Matsuo : Effect of Acetazolamide on Osteoclasts and Osteoblasts in the rat, The International Association for Dental Research, Orlando(USA), Mar. 1997.
81.
Shigeyuki Ebisu, T Yasunaga, Toshiyuki Suge, Akiko Kawasaki, Kunio Ishikawa, Kenzo Asaoka and Takashi Matsuo : Acid resistance of bovine enamel after calcium phosphate precipitation method using dogs, Journal of Dental Research, Vol.76, No.Special Issue, 180, Orlando, Mar. 1997.
82.
Akiko Kawasaki, Kunio Ishikawa, Toshiyuki Suge, Kenzo Asaoka, Takashi Matsuo and Shigeyuki Ebisu : Effects of plaque control on the occlusion of dentin tubules, Journal of Dental Research, Vol.76, No.Special Issue, 180, Orlando, Mar. 1997.
83.
Toshiyuki Suge, Kunio Ishikawa, Akiko Kawasaki, Kenzo Asaoka, Takashi Matsuo and Shigeyuki Ebisu : Acid resistance of bovine enamel after calcium phosphate precipitation method, Journal of Dental Research, Vol.76, No.Special Issue, 97, Orlando, Mar. 1997.
84.
Kazumi Ozaki, Yuichirou Noiri, Hideaki Nakae, Takashi Matsuo and Shigeyuki Ebisu : A histopathological and microbiological study on experimental root-surface caries in humans, 74th General Session of IADR Meeting, San Francisco, Mar. 1996.
85.
Yuichirou Noiri, Kazumi Ozaki, Tae Okamoto, Keiko Fujinaka, Takashi Matsuo and Shigeyuki Ebisu : Localization of periodontal disease-associated bacteria in human periodontal pockets, 74th General Session of IADR Meeting, San Francisco, Mar. 1996.
86.
H Okada, Y Shimabukuro, Y Kassai, Hiro-O Ito, Takashi Matsuo, S Ebisu and Y Harada : The Function of Gingival Lymphocytes on the Establishment of Human Periodontitis, Advances in Dental Research, Vol.2, No.2, 364-367, Kobe, Nov. 1988.
Nur Asikin, Katsuhiko Hirota, Hiromichi Yumoto, Kouji Hirao, Kanako Takahashi, Keiji Murakami, Takashi Matsuo and Yoichiro Miyake : Effect of extracellular DNA on pyocyanin production from Pseudomonas aeruginosa, 第27回Bacterial Agherence & Biofilm学術集会, Jul. 2013.
Nur Asikin, Katsuhiko Hirota, Hiromichi Yumoto, Kouji Hirao, Kanako Takahashi, Keiji Murakami, Takashi Matsuo and Yoichiro Miyake : Role of Extracellular DNA and DNA-binding protein in biofilm formation of Streptococcus intermeius, 第26回Bacterial Adherence & Biofilm学術集会, Jul. 2012.
Nur Asikin, Katsuhiko Hirota, Hiromichi Yumoto, Kouji Hirao, Kanako Takahashi, Keiji Murakami, Takashi Matsuo and Yoichiro Miyake : Roles of extracellular DNA and DNA-binding protein in cell aggregation, biofilm formation and antibiotic tolerance of Streptococcus intermedius, 第21回Lancefieldレンサ球菌研究会&第44回レンサ球菌感染症研究会合同開催, Jun. 2012.
Ikuko Hosokawa, Yoshitaka Hosokawa, Kazumi Ozaki, Hideaki Nakae and Takashi Matsuo : Adrenomedullin inhibits CXCL10 production by human gingival fibroblasts, 56th Annual Meeting of Japanese Association for Dental Research, Nov. 2008.
Liu Dali, Hiromichi Yumoto, Keiji Murakami, Katsuhiko Hirota, Shizuo Kayama, Takashi Matsuo and Yoichiro Miyake : The essentiality and involvements of Streptococcus intermedius histone-like protein in bacterial viability and normal growth, The 2nd international symposium on "The future direction of oral sciences in the 21st century", Dec. 2007.
Ikuko Hosokawa, Yoshitaka Hosokawa, Kazumi Ozaki, Hiromichi Yumoto, Hideaki Nakae and Takashi Matsuo : Proinflammatory Roles of NOD2 in Human Gingival Fibroblasts, 第55回 国際歯科研究学会日本部会(JADR)総会・学術大会, Nov. 2007.
98.
Ikuko Hosokawa, Yoshitaka Hosokawa, Kazumi Ozaki, Hiromichi Yumoto, Hideaki Nakae and Takashi Matsuo : Proinflammatory Roles of NOD2 in Human Gingival Fibroblasts, 第55回 国際歯科研究学会日本部会(JADR)総会・学術大会, Nov. 2007.
99.
Dali LIU, 湯本 浩通, 松尾 敬志, 三宅 洋一郎 : Histone-like DNA binding protein of Streptococcus intermedius induces the expression of pro-inflammatory cytokines in human monocytes via activation of ERK1/2 and JNK pathways., 日本歯周病学会50周年記念大会, 2007年9月.
Akiko Kawasaki, Kunio Ishikawa, Toshiyuki Suge, Kenzo Asaoka, Takashi Matsuo and Shigeyuki Ebisu : Effect of apatite forming method on the crystallinity of hydroxyapatite powder, Transaction of the 12 th Symposium on apatite, 6-7, Dec. 1996.
191.
Toshiyuki Suge, Kunio Ishikawa, Akiko Kawasaki, Kenzo Asaoka, Takashi Matsuo and Shigeyuki Ebisu : Apatite forming method -duration of dentinal tubules occlusion-, Transactions of the 12th Symposium on apatite, 4-5, Dec. 1996.
192.
Toshiyuki Suge, Kunio Ishikawa, Akiko Kawasaki, Kenzo Asaoka, Takashi Matsuo and Shigeyuki Ebisu : A new treatment method for dentin hypersensitivity -apatite forming method-, Transactions of the 12th Symposium on apatite, 2-3, Dec. 1996.
尾崎 和美, 松尾 敬志 : Bacterial Viability Assay at an Electron Microscopic Leved, カリオロージに基づく齲蝕の診断法および治療法の開発, 51-58, 2005年3月.
4.
尾崎 和美, 松尾 敬志 : PCR Amplificcation of Target DNA Extracted From Bacterial Cells Embedded in Plastic Material for Electron Microscopy, カリオロージに基づく齲蝕の診断法および治療法の開発, 29-49, 2005年3月.
(キーワード)
Bactrerial DNA / Semiquantitative PCR / Embedding-Material