Yuji Inagaki, Jun-ichi Kido, Yasufumi Nishikawa, Rie Kido, Eijiro Sakamoto, Mika Bandou, Koji Naruishi, Toshihiko Nagata and Hiromichi Yumoto : Gan-Lu-Yin (Kanroin), Traditional Chinese Herbal Extracts, Reduces Osteoclast Differentiation In Vitro and Prevents Alveolar Bone Resorption in Rat Experimental Periodontitis, Jul. 2021.
Academic Paper (Judged Full Paper):
1.
Rie Kido, Yuka Hiroshima, Jun-ichi Kido, Mika Bandou, Kaya Yoshida, Yuji Inagaki and Hiromichi Yumoto : Lipocalin 2 inhibits the expressions of interleukin-8 and macrophage inflammatory protein-1α in human neutrophil-like cells, Journal of Oral Biosciences, Vol.67, No.1, 2025.
(Summary)
Lipocalin 2 (LCN2) is a glycoprotein with multiple functions, including antimicrobial activity, inflammatory response modulation, and cell migration. LCN2 is expressed in some cells, such as epithelial cells and neutrophils, and its levels are increased in inflammatory diseases. This study investigated the presence of LCN2 receptor (24p3R) in cells around periodontal tissues and function of LCN2 in cells with a receptor to explore the role of LCN2 in periodontal diseases. The presence of 24p3R was examined in periodontal cells, including human gingival fibroblasts, periodontal ligament fibroblasts, human oral epithelial cells (HOECs), and neutrophil-like cells (HL-60), by Western blotting. Changes in periodontal disease-associated proteins in the presence of recombinant LCN2 (rLCN2) were examined using a protein array in differentiated HL-60 (dHL-60) cells. Interleukin-8 (IL-8) and macrophage inflammatory protein-1α (MIP-1α) mRNA expressions were analyzed by qRT-PCR, and IL-8 and MIP-1α levels in dHL-60 cells treated with rLCN2 or Porphyromonas gingivalis-lipopolysaccaharide (P.g-LPS) were determined by enzyme-linked immunosorbent assay. We detected 24p3R in dHL-60 cells. IL-8 was highly expressed and MIP-1α was weakly expressed in dHL-60 cells using a protein array. rLCN2 significantly decreased IL-8 mRNA and protein levels and suppressed P.g-LPS-induced IL-8 production in dHL-60 cells. As dHL-60 cells were co-cultured with HOECs in which LCN2 was knocked down, IL-8 mRNA expression increased in dHL-60 cells. Furthermore, rLCN2 inhibited MIP-1α production in dHL-60 cells. LCN2 suppresses inflammatory responses by regulating IL-8 and MIP-1α expression in periodontal diseases.
Kenji Masutomi, Mika Bando, Yuji Inagaki, Rie Kido, Yuta Uemura, Yukari Hatada, Ichi Jun Kido, Makoto Fukui, Daisuke Hinode and Hiromichi Yumoto : Relationship between oral hypofunction and salivary biomarkers in older adults: a cross-sectional study, BMC Oral Health, Vol.24, No.1, 766, 2024.
(Summary)
Oral health problems have increased among older adults. Oral hypofunction is characterized by seven signs and symptoms: oral uncleanness, oral dryness, decline in occlusal force, decline in the movement function of the tongue and lips, decline in tongue pressure, decline in masticatory function, and decline in swallowing function, the latter being a significant risk factors for oral frailty. Recent research has suggested that salivary biomarkers can be used to assess not only oral diseases, including dental caries and periodontitis, but also systemic diseases, such as cancer and diabetes mellitus. This cross-sectional study investigated the relationship between oral hypofunction and the levels of salivary biomarkers. In total, 116 patients, aged 65 years or older, were included in this cross-sectional study. If three or more signs or symptoms in seven kinds of tests met the criteria of each test, oral hypofunction was diagnosed. The levels of biomarkers in the saliva collected from the patients were analyzed using an enzyme-linked immunosorbent assay. In total, 63.8% of patients were diagnosed with oral hypofunction. Multivariable linear regression analysis showed that calprotectin levels in the saliva were significantly related to oral moisture and masticatory function. Furthermore, 8-OHdG levels in saliva were associated with the movement function of the tongue and lips and oral hygiene level, and salivary AGE correlated only with the movement function of the tongue and lips. Multiple logistic regression analysis revealed that calprotectin levels in the saliva were significantly correlated with the prevalence of oral hypofunction, even after adjusting for age, sex, and periodontal status. However, none of the biomarker levels in the saliva had a significant relationship with the number of examinations outside the reference range. Calprotectin, 8-OHdG, and AGE levels are associated with oral hypofunction in older adults.
Yuka Hiroshima, Rie Kido, Jun-ichi Kido, Mika Bandou, Kaya Yoshida, Akikazu Murakami and Yasuo Shinohara : Synthesis of secretory leukocyte protease inhibitor using cell-free protein synthesis system, Odontology, Vol.112, No.4, 1103-1112, 2024.
(Summary)
Secretory leukocyte protease inhibitor (SLPI) functions as a protease inhibitor that modulates excessive proteolysis in the body, exhibits broad-spectrum antimicrobial activity, regulates inflammatory responses, and plays an important role in the innate immunity. The purpose of the study was to artificially synthesize a SLPI, an antimicrobial peptide, and investigate its effect on antimicrobial activity against Porphyromonas gingivalis and interleukin-6 (IL-6) production. SLPI protein with a molecular weight of approximately 13 kDa was artificially synthesized using a cell-free protein synthesis (CFPS) system and investigated by western blotting and enzyme-linked immunosorbent assay (ELISA). Disulfide bond isomerase in the protein synthesis mixture increased the amount of SLPI synthesized. The synthesized SLPI (sSLPI) protein was purified and its antimicrobial activity was investigated based on the growth of Porphyromonas gingivalis and bacterial adhesion to oral epithelial cells. The effect of sSLPI on IL-6 production in human periodontal ligament fibroblasts (HPLFs) was examined by ELISA. Our results showed that sSLPI significantly inhibited the growth of Porphyromonas gingivalis and bacterial adhesion to oral epithelial cells and further inhibited IL-6 production by HPLFs. These results suggested that SLPI artificially synthesized using the CFPS system may play a role in the prevention of periodontal diseases through its antimicrobial and anti-inflammatory effects.
Jun-ichi Kido, Yuka Hiroshima, Rie Kido, Kaya Yoshida, Yuji Inagaki, Koji Naruishi, Kazuaki Kajimoto, Masatoshi Kataoka, Yasuo Shinohara and Hiromichi Yumoto : Lipocalin 2, synthesized using a cell-free protein synthesis system and encapsulated into liposomes, inhibits the adhesion of Porphyromonas gingivalis to human oral epithelial cells., Journal of Periodontal Research, Vol.58, No.2, 262-273, 2023.
Yuka Hiroshima, Jun-ichi Kido, Rie Kido, Kaya Yoshida, Mika Bandou, Kazuaki Kajimoto, Hiromichi Yumoto and Yasuo Shinohara : β-defensin 2 synthesized by a cell-free protein synthesis system and encapsulated in liposomes inhibits adhesion of Porphyromonas gingivalis to oral epithelial cells., Odontology, Vol.111, 830-838, 2023.
Rie Kido, Jun-ichi Kido, Yasufumi Nishikawa, Eijiro Sakamoto, Yoritoki Tomotake and Hiromichi Yumoto : Diagnosis of inflammatory peri-implant diseases using an immunochromatographic assay for calprotectin in peri-implant crevicular fluid, International Journal of Implant Dentistry, Vol.7, No.1, 106, 2021.
(Summary)
The incidence rate of peri-implant diseases is increasing with implant placement. Early detection of peri-implant diseases is important to prevent and treat these diseases, and a simple and objective diagnostic method is expected. Immunochromatographic (IC) assays are used for rapid diagnostic methods for some diseases. The aim of this clinical study was to determine the amount of calprotectin, an inflammatory marker, in peri-implant crevicular fluid (PICF) using an IC chip, and estimate the possibility of this diagnostic system. Forty-six individuals with dental implants participated in a pilot study. PICF samples were collected from the peri-implant sites with or without inflammation after clinical examinations including probing depth (PD), bleeding on probing (BOP) and gingival index (GI). Calprotectin in PICF was determined by an IC chip and enzyme-linked immunosorbent assay (ELISA) for calprotectin. The density of calprotectin line on the IC chip was measured using an IC reader (IC reader value). The relationship between IC reader value and ELISA value or clinical parameters was investigated. A receiver operating characteristic (ROC) curve analysis of IC reader value of calprotectin was performed to predict inflammation in peri-implant diseases. IC reader value of calprotectin was significantly correlated with its ELISA value and PD. IC reader values of calprotectin in PICF samples from periodontal sites with GI-1 and GI-2, and with BOP-positive sites were significantly higher than those of PICF samples from GI-0 sites, and BOP-negative sites, respectively. The IC reader value for calprotectin in PICF samples from inflammatory diseased sites was significantly higher than that of non-diseased sites. ROC analysis suggested that the IC reader value of PICF calprotectin was useful for predicting inflammatory peri-implant diseases. IC assay for PICF calprotectin may be a possible system for diagnosing the inflammatory peri-implant diseases.
Yuji Inagaki, Jun-ichi Kido, Yasufumi Nishikawa, Rie Kido, Eijiro Sakamoto, Mika Bandou, Koji Naruishi, Toshihiko Nagata and Hiromichi Yumoto : Gan-Lu-Yin (Kanroin), traditional Chinese herbal extracts, reduces osteoclast differentiation in vitro and prevents alveolar bone resorption in rat experimental periodontitis, Journal of Clinical Medicine, Vol.10, No.3, 386, 2021.
(Summary)
Gan-Lu-Yin (GLY), a traditional Chinese herbal medicine, shows therapeutic effects on periodontitis, but that mechanism is not well known. This study aims to clarify the precise mechanism by investigating the inhibitory effects of GLY extracts on osteoclastogenesis in vitro and on bone resorption in periodontitis in vivo. RAW264.7 cells are cultured with soluble receptor activator of nuclear factor-kappa B (sRANKL) and GLY extracts (0.01-1.0 mg/mL), and stained for tartrate-resistant acid phosphatase (TRAP) to evaluate osteoclast differentiation. Experimental periodontitis is induced by placing a nylon ligature around the second maxillary molar in rats, and rats are administered GLY extracts (60 mg/kg) daily for 20 days. Their maxillae are collected on day 4 and 20, and the levels of alveolar bone resorption and osteoclast differentiation are estimated using micro-computed tomography (CT) and histological analysis, respectively. In RAW264.7 cells, GLY extracts significantly inhibit sRANKL-induced osteoclast differentiation at a concentration of more than 0.05 mg/mL. In experimental periodontitis, administering GLY extracts significantly decreases the number of TRAP-positive osteoclasts in the alveolar bone on day 4, and significantly inhibits the ligature-induced bone resorption on day 20. These results show that GLY extracts suppress bone resorption by inhibiting osteoclast differentiation in experimental periodontitis, suggesting that GLY extracts are potentially useful for oral care in periodontitis.
Koji Naruishi, Eijiro Sakamoto, Takahisa Ikuta, Rie Kido, Jun-ichi Kido and Hiromichi Yumoto : Evaluation of educational effects of periodontology by lectures and phantom training in dental students, The Japanese Journal of Conservative Dentistry, Vol.63, No.1, 22-29, 2020.
(Summary)
<p> Purpose: In the undergraduate educational curriculum of dentistry, there are two main teaching/learning methods for dental students before clinical training: traditional lectures and phantom training. The purpose of this study was to explore effective educational systems for periodontology by examining the test results of both lectures and phantom training.</p><p> Methods: Both a written test and a practical test of periodontology were administered to students of dentistry. A total of 43 dental students of Tokushima University Dental School were recruited. Written tests were performed before and after the lectures on periodontology. A practical test of scaling and root planing was performed after phantom training. Associations between the results of the written test and practical test were analyzed statistically. P values of less than 0.05 were considered statistically significant.</p><p> Results: The results of the written tests after lectures were higher than those before lectures. A significant positive correlation was found between the results of the written test and practical test.</p><p> Conclusion: In the students' education on periodontology, the results of the practical test were correlated statistically to the results of the written test. These findings may contribute to establishing effective educational systems for periodontology in dentistry.</p>
Eijiro Sakamoto, Rie Kido, Yoritoki Tomotake, Yoshihito Naitou, Yuichi Ishida and Jun-ichi Kido : Calprotectin and cross-linked N-telopeptides of type I collagen levels in crevicular fluid from implant sites with peri-implant diseases: a pilot study, International Journal of Implant Dentistry, Vol.4, No.1, 26, 2018.
(Summary)
Peri-implant crevicular fluid (PICF) contains calprotectin and NTx, which are markers for inflammation and bone resorption, respectively. The aims of this pilot study were to compare calprotectin and NTx levels in PICF from implant sites with or without peri-implant diseases and to evaluate the usefulness of calprotectin and NTx as diagnostic markers for peri-implant diseases. Thirty-five patients with dental implants participated in this pilot study. PICF samples were collected from peri-implant disease sites (n = 40) and non-diseased (healthy) sites (n = 34) after clinical indicators including probing depth (PD), bleeding on probing (BOP), gingival index (GI), and bone loss (BL) rate were investigated. Calprotectin and NTx amounts in PICF were measured using their respective ELISA kits and then compared between diseased and healthy samples. The relationship between PICF calprotectin or NTx levels and clinical indicator levels was investigated. A receiver operating characteristic (ROC) curve analysis of calprotectin and NTx was performed to predict peri-implant diseases. Calprotectin and NTx levels in PICF were significantly higher from peri-implant disease sites than from healthy sites. PICF calprotectin amounts correlated with PD, and its levels were significantly higher in the GI-1 and GI-2 groups than in the GI-0 group. PICF NTx amounts correlated with PD and the BL rate. ROC curves indicated that PICF calprotectin and NTx are useful biomarkers for peri-implant diseases. Calprotectin and NTx in PICF have potential as biomarkers for the diagnosis of peri-implant diseases.
Takeo Iwata, Kyoko Kuribayashi, Masahiko Nakasono, Noriko Saito-Tarashima, Noriaki Minakawa, Noriko Mizusawa, Rie Kido and Katsuhiko Yoshimoto : The AMPK/mTOR pathway is involved in D-dopachrome tautomerase gene transcription in adipocytes differentiated from SGBS cells, a human preadipocyte cell line, Cytokine, Vol.96, 195-202, 2017.
(Summary)
In adipose tissue, D-dopachrome tautomerase (DDT), a cytokine with structural similarity to macrophage migration inhibitory factor, is mainly expressed in adipocytes rather than preadipocytes and acts as an anti-obesity adipokine in an autocrine manner. However, its transcriptional regulation is largely unknown. In order to explore molecules affecting DDT transcription, a chemical library screening using HEK293 cells stably expressing a DDT promoter-reporter construct was performed. Several derivatives of 5-aminoimidazole-4-carboxamide-1--d-ribofuranoside (AICAR), an AMP-activated protein kinase (AMPK) activator, were identified as transcriptional activators of the DDT gene. Furthermore, DDT mRNA levels were reduced in SGBS adipocytes treated with compound C, an AMPK inhibitor, suggesting involvement of AMPK in DDT transcription. Overexpression of the FOXO1 constitutive active form reduced transcriptional activity of the DDT gene in SGBS cells, but increased it in HEK293 cells. Cell-type specific effects were also observed in the DDT gene expression of cells treated with AS1842856, a FOXO1 inhibitor. Finally, involvement of the mammalian target of rapamycin (mTOR) signaling in DDT transcription in SGBS adipocytes was investigated. Rapamycin, an inhibitor of mTOR, increased DDT mRNA levels and attenuated the inhibitory effects of compound C on DDT mRNA levels in SGBS adipocytes. In conclusion, DDT transcription may be regulated in a cell-dependent manner, and were enhanced by AMPK activation in SGBS adipocytes through inhibiting the mTOR signaling.
Eijiro Sakamoto, Yuka Hiroshima, Jun-ichi Kido, Yasufumi Nishikawa, Koji Naruishi, Rie Kido and Hiromichi Yumoto : Various roles of calprotectin in periodontal disease and its possibility as a diagnostic marker for periodontal disease, Journal of the Japanese Association of Periodontology, Vol.62, No.4, 193-199, Dec. 2020.
枡富 健二, Mika Bandou, Yuji Inagaki, Rie Kido, Yuta Uemura, 畑田 祐佳里, Jun-ichi Kido and Hiromichi Yumoto : 歯周病高齢患者のオーラルフレイルと口腔内バイオマーカーとの関連, 第66回秋季日本歯周病学会学術大会, Oct. 2023.
3.
畑田 祐佳里, Yuji Inagaki, 仲村 大輔, Yuta Uemura, Rie Kido and Hiromichi Yumoto : 藍(タデアイ)に含まれるTryptanthrinは破骨細胞の分化を抑制する, 第66回秋季日本歯周病学会学術大会, Oct. 2023.
4.
Mika Bandou, 枡富 健二, Yuji Inagaki, Rie Kido, Yuta Uemura, 畑田 祐佳里, Jun-ichi Kido and Hiromichi Yumoto : 糖尿病関連歯周炎高齢者患者のオーラルフレイルリスクバイオマーカーの探索, 四国歯学会第62回例会, Jul. 2023.
5.
枡富 健二, Mika Bandou, Yuji Inagaki, Rie Kido, 植村 勇太, 畑田 祐佳里, Jun-ichi Kido and Hiromichi Yumoto : 糖尿病関連歯周炎高齢患者のオーラルフレイルリスクバイオマーカーの探索, 第66回春季日本歯周病学会学術大会(高松), May 2023.
6.
Rie Kido, Yuka Hiroshima, Jun-ichi Kido, Kaya Yoshida, Takahisa Ikuta and Hiromichi Yumoto : Antimicrobial activity of artificially synthesized secretory leukocyte protease inhibitor and its encapsulation into liposomes, 第66回春季日本歯周病学会学術大会, May 2023.
7.
Jun-ichi Kido, Yuka Hiroshima, Rie Kido, Kaya Yoshida, Yuji Inagaki, Koji Naruishi and Hiromichi Yumoto : 人工合成したβ-defensin 2によるPorphyromonas gingivalisの付着抑制およびリポソーム封入と口腔上皮細胞への送達, 四国歯学会第60回例会, Jun. 2022.
8.
Jun-ichi Kido, Yuka Hiroshima, Rie Kido, Kaya Yoshida, Yuji Inagaki, Koji Naruishi and Hiromichi Yumoto : 人工合成したbeta-defensin 2によるPorphyromonas gingivalisの付着抑制およびリポソーム封入と口腔上皮細胞への送達, 第65回春季日本歯周病学会学術大会, Jun. 2022.
9.
Rie Kido, Eijiro Sakamoto, Yuka Hiroshima, Mika Bandou, Jun-ichi Kido, Takahisa Ikuta, Masami Ninomiya, Yuji Inagaki, Yasufumi Nishikawa, Yoritoki Tomotake, Koji Naruishi and Hiromichi Yumoto : イムノクロマト法を用いたカルプロテクチン測定による歯周病およびインプラント疾患の診断, 四国歯学会第59回例会, Mar. 2022.
10.
Yuka Hiroshima, Rie Kido, Jun-ichi Kido, Yuji Inagaki, Koji Naruishi and Hiromichi Yumoto : 人工合成したLopocalin 2はPorphyromonas gingivalisの口腔上皮細胞への付着を抑制する, 第64回秋季日本歯周病学会学術大会, Oct. 2021.
11.
Jun-ichi Kido, Yuka Hiroshima, Rie Kido, Kaya Yoshida, Yuji Inagaki, Koji Naruishi and Hiromichi Yumoto : リポソームに封入したリポカリン2の口腔上皮細胞への送達, 第64回春季日本歯周病学会学術大会, May 2021.
Jun-ichi Kido, Yuka Hiroshima, Rie Kido, Yuji Inagaki, Koji Naruishi and Hiromichi Yumoto : 無細胞蛋白質合成系を用いた抗菌ペプチドの合成とリポソーム封入, 日本歯科保存学会2020年度春季学術大会(第152回)(神戸), Jun. 2020.
14.
Yuka Hiroshima, Jun-ichi Kido, Rie Kido, Kaya Yoshida, Yuji Inagaki, Koji Naruishi and Hiromichi Yumoto : オーラルケアへの応用に関連するリポソームのデリバリー法の検討, 第63回春季日本歯周病学会学術大会, May 2020.
15.
Rie Kido, Yuka Hiroshima, Takahisa Ikuta, Yuji Inagaki, Mika Bandou, Koji Naruishi, Jun-ichi Kido and Hiromichi Yumoto : 最終糖化産物による口腔上皮細胞の Lipocalin2 発現誘導は好中球の遊走性とサイトカイン発現を 調節する, 第63回春季日本歯周病学会学術大会, May 2020.