Tsuyoshi Morita, Shin Matsumoto and Otto Baba : Expression of secretory calcium-binding phosphoprotein (scpp) genes in medaka during the formation and replacement of pharyngeal teeth, BMC Oral Health, Vol.23, No.1, 744, 2023.
(要約)
Analyses of tooth families and tooth-forming units in medaka with regard to tooth replacement cycles and the localization of odontogenic stem cell niches in the pharyngeal dentition clearly indicate that continuous tooth replacement is maintained. The secretory calcium-binding phosphoprotein (scpp) gene cluster is involved in the formation of mineralized tissues, such as dental and bone tissues, and the genes encoding multiple SCPPs are conserved in fish, amphibians, reptiles, and mammals. In the present study, we examined the expression patterns of several scpp genes in the pharyngeal teeth of medaka to elucidate their roles during tooth formation and replacement. Himedaka (Japanese medaka, Oryzias latipes) of both sexes (body length: 28 to 33 mm) were used in this study. Real-time quantitative reverse transcription-polymerase chain reaction (PCR) (qPCR) data were evaluated using one-way analysis of variance for multi-group comparisons, and the significance of differences was determined by Tukey's comparison test. The expression of scpp genes was examined using in situ hybridization (ISH) with a digoxigenin-labeled, single-stranded antisense probe. qPCR results showed that several scpp genes were strongly expressed in pharyngeal tissues. ISH analysis revealed specific expression of scpp1, scpp5, and sparc in tooth germ, and scpp5 was continually expressed in the odontoblasts of teeth attached to pedicles, but not in the osteoblasts of pedicles. In addition, many scpp genes were expressed in inner dental epithelium (ide), but not in odontoblasts, and scpp2 consistently showed epithelial-specific expression in the functional teeth. Taken together, these data indicate that specific expression of scpp2 and scpp5 may play a critical role in pharyngeal tooth formation in medaka. We characterized changes in the expression patterns of scpp genes in medaka during the formation and replacement of pharyngeal teeth.
(キーワード)
Humans / Animals / Oryzias / Calcium / Phosphoproteins / Odontogenesis / Bone and Bones / Mammals
Katsuhiko Ono, Hitoshi Gotoh, Tadashi Nomura, Tsuyoshi Morita, Otto Baba, Mami Matsumoto, Sei Saitoh and Nobuhiko Ohno : Ultrastructural characteristics of oligodendrocyte precursor cells in the early postnatal mouse optic nerve observed by serial block-face scanning electron microscopy, PLoS ONE, Vol.17, No.12, 1-18, 2022.
(要約)
Oligodendrocyte precursor cells (OPC) arise from restricted regions of the central nervous system (CNS) and differentiate into myelin-forming cells after migration, but their ultrastructural characteristics have not been fully elucidated. This study examined the three-dimensional ultrastructure of OPCs in comparison with other glial cells in the early postnatal optic nerve by serial block-face scanning electron microscopy. We examined 70 putative OPCs (pOPC) that were distinct from other glial cells according to established morphological criteria. The pOPCs were unipolar in shape with relatively few processes, and their Golgi apparatus were localized in the perinuclear region with a single cisterna. Astrocytes abundant in the optic nerve were distinct from pOPCs and had a greater number of processes and more complicated Golgi apparatus morphology. All pOPCs and astrocytes contained a pair of centrioles (basal bodies). Among them, 45% of pOPCs extended a short cilium, and 20% of pOPCs had centrioles accompanied by vesicles, whereas all astrocytes with basal bodies had cilia with invaginated ciliary pockets. These results suggest that the fine structures of pOPCs during the developing and immature stages may account for their distinct behavior. Additionally, the vesicular transport of the centrioles, along with a short cilium length, suggests active ciliogenesis in pOPCs.
Arief Waskitho, Yumiko Yamamoto, S Raman, Fumiya Kano, Huijiao Yan, R Raju, S Afroz, Tsuyoshi Morita, Daisuke Ikutame, Kazuo Okura, Masamitsu Ohshima, Akihito Yamamoto, Otto Baba and Yoshizo Matsuka : Peripherally Administered Botulinum Toxin Type A Localizes Bilaterally in Trigeminal Ganglia of Animal Model, Toxins, Vol.13, No.10, 704, 2021.
Huijiao Yan, Masamitsu Ohshima, Raju Raju, S Raman, Kazumitsu Sekine, Arief Waskitho, Miho Inoue, Masahisa Inoue, Otto Baba, Tsuyoshi Morita, Mayu Miyagi and Yoshizo Matsuka : Dentin-pulp complex tissue regeneration via three-dimensional cell sheet layering, Tissue Engineering. Part C, Methods, Vol.27, No.10, 559-570, 2021.
(要約)
The dentin-pulp complex is a unique structure in teeth that contains both hard and soft tissues. Generally, deep caries and trauma cause damage to the dentin-pulp complex, and if left untreated, this damage will progress to irreversible pulpitis. The aim of this study was to fabricate a layered cell sheet composed of rat dental pulp (DP) cells and odontogenic differentiation of pulp (OD) cells and to investigate the ability to regenerate the dentin-pulp complex in a scaffold tooth. We fabricated two single cell sheets composed of DP cells (DP cell sheet) or OD cells (OD cell sheet) and a layered cell sheet made by layering both cells. The characteristics of the fabricated cell sheets were analyzed using light microscopy, scanning electron microscope (SEM), hematoxylin-eosin (HE) staining, and immunohistochemistry (IHC). Furthermore, the cell sheets were transplanted into the subrenal capsule of immunocompromised mice for 8 weeks. After this, the regenerative capacity to form dentin-like tissue was evaluated using micro-computed tomography (micro-CT), HE staining, and IHC. The findings of SEM and IHC confirmed that layered cell sheets fabricated by stacking OD cells and DP cells maintained their cytological characteristics. Micro-CT of layered cell sheet transplants revealed a mineralized capping of the access cavity in the crown area, similar to that of natural dentin. In contrast, the OD cell sheet group demonstrated the formation of irregular fragments of mineralized tissue in the pulp cavity, and the DP cell sheet did not develop any hard tissue. Moreover, bone volume/tissue volume (BV/TV) showed a significant increase in hard tissue formation in the layered cell sheet group compared with that in the single cell sheet group ( < 0.05). HE staining also showed a combination of soft and hard tissue formation in the layered cell sheet group. Furthermore, IHC confirmed that the dentin-like tissue generated from the layered cell sheet expressed characteristic markers of dentin but not bone equivalent to that of a natural tooth. In conclusion, this study demonstrates the feasibility of regenerating dentin-pulp complex using a bioengineered tissue designed to simulate the anatomical structure. Impact statement The dentin-pulp complex can be destroyed by deep caries and trauma, which may cause pulpitis and progress to irreversible pulpitis, apical periodontitis, and even tooth loss. Current treatments cannot maintain pulp health, and teeth can become brittle. We developed a three-dimensional (3D) layered cell sheet using dental pulp cells and odontogenic differentiation of pulp cells for dentin-pulp complex regeneration. Our layered cell sheet enables the regeneration of an organized 3D dentin-pulp-like structure comparable with that of natural teeth. This layered cell sheet technology may contribute to dentin-pulp complex regeneration and provide a novel method for complex tissue engineering.
Han Zhu, Zhang Weizhi, Ning Wanshan, Wang Chenwei, Deng Wankun, Li Zhidan, Shang Zehua, Shen Xiaofei, Liu Xiaohui, Otto Baba, Tsuyoshi Morita, Chen Lu, Xue Yu and Jia Da : Model-based analysis uncovers mutations altering autophagy selectivity in human cancer, Nature Communications, Vol.12, No.3258, 1-18, 2021.
(要約)
Autophagy can selectively target protein aggregates, pathogens, and dysfunctional organelles for the lysosomal degradation. Aberrant regulation of autophagy promotes tumorigenesis, while it is far less clear whether and how tumor-specific alterations result in autophagic aberrance. To form a link between aberrant autophagy selectivity and human cancer, we establish a computational pipeline and prioritize 222 potential LIR (LC3-interacting region) motif-associated mutations (LAMs) in 148 proteins. We validate LAMs in multiple proteins including ATG4B, STBD1, EHMT2 and BRAF that impair their interactions with LC3 and autophagy activities. Using a combination of transcriptomic, metabolomic and additional experimental assays, we show that STBD1, a poorly-characterized protein, inhibits tumor growth via modulating glycogen autophagy, while a patient-derived W203C mutation on LIR abolishes its cancer inhibitory function. This work suggests that altered autophagy selectivity is a frequently-used mechanism by cancer cells to survive during various stresses, and provides a framework to discover additional autophagy-related pathways that influence carcinogenesis.
Tsuyoshi Kano, Tsuyoshi Morita, Kaori Sumida, Hiromichi Yumoto and Otto Baba : Expression of fibroblast growth factor receptor1, -2c, and -3c transcripts in mouse molars after tooth eruption, Anatomical Science International, 2021.
Resmi Raju, Masamitsu Ohshima, Miho Inoue, Tsuyoshi Morita, Yan Huijiao, Arief Waskitho, Otto Baba, Masahisa Inoue and Yoshizo Matsuka : Three-dimensional periodontal tissue regeneration using a bone-ligament complex cell sheet, Scientific Reports, Vol.10, No.1, 1656, 2020.
(要約)
Periodontal tissue is a distinctive tissue structure composed three-dimensionally of cementum, periodontal ligament (PDL) and alveolar bone. Severe periodontal diseases cause fundamental problems for oral function and general health, and conventional dental treatments are insufficient for healing to healthy periodontal tissue. Cell sheet technology has been used in many tissue regenerations, including periodontal tissue, to transplant appropriate stem/progenitor cells for tissue regeneration of a target site as a uniform tissue. However, it is still difficult to construct a three-dimensional structure of complex tissue composed of multiple types of cells, and the transplantation of a single cell sheet cannot sufficiently regenerate a large-scale tissue injury. Here, we fabricated a three-dimensional complex cell sheet composed of a bone-ligament structure by layering PDL cells and osteoblast-like cells on a temperature responsive culture dish. Following ectopic and orthotopic transplantation, only the complex cell sheet group was demonstrated to anatomically regenerate the bone-ligament structure along with the functional connection of PDL-like fibers to the tooth root and alveolar bone. This study represents successful three-dimensional tissue regeneration of a large-scale tissue injury using a bioengineered tissue designed to simulate the anatomical structure.
Kikuji Yamashita, Dalkhsuren Shine-Od, Aldartsogt Dolgorsuren, Kaori Sumida, Tsuyoshi Morita and Otto Baba : Effects of Dietary Cure Containing AFIR (Antioxidant Far-Infrared Energy Radiating) Ceramics on Living Body of Hatchery Sea Bream (Pagrus Major), International Journal of Innovative Studies in Sciences and Engineering Technology, Vol.2, No.10, 7-16, 2016.
9.
Tsuyoshi Morita, Kaoru Fujikawa, Otto Baba and Shibata Shunichi : An in situ hybridization study of Hyaluronan synthase (Has) mRNA in developing mouse molar and incisor tooth germs, Gene Expression Patterns, Vol.21, No.1, 28-40, 2016.
(要約)
Hyaluronan (HA) is a major constituent molecule in most extracellular matrices and is synthesized by Hyaluronan synthase (Has). In the present study, we examined expression patterns of Has1, -2, -3 mRNA in developing mouse molar and incisor tooth germs from embryonic day (E) 11.5 to postnatal day (P) 7, focusing on Hertwig's epithelial root sheath (HERS) and the apical bud in particular. Has1 mRNA expression was not detected in all tooth germs examined. Has2 mRNA was expressed in the surrounding mesenchyme from E12.0 to 18.0 in both molar and incisor tooth germs, but disappeared after birth. Meanwhile, Has3 mRNA was exclusively expressed within the enamel organ, especially in the inner enamel epithelium (IEE), stellate reticulum (SR), and stratum intermedium (SI) until the early bell stage at E16.0. Has3 mRNA disappeared as IEE differentiated into differentiating ameloblasts (dABs), but remained in SI until the root developmental stage of the molar tooth germ at P7. Has3 mRNA was also expressed in HERS until P7. In incisors, Has3 mRNA was expressed in the apical bud, especially in the transit-amplifying (TA) cell region from E16.0 to P7, and in the papillary layer (PL) adjacent to the mature enamel. These gene expression patterns suggested that Has3 is the main control factor for prenatal and postnatal HA synthesis of the tooth germ, and may in part regulate crown and root formation of the tooth germ, maintenance of stem cell niches in the apical bud as well as mineral transport in PL.
R Raju, Masamitsu Ohshima, Tsuyoshi Morita, Huijiao Yan, Miho Inoue, Otto Baba, Masahisa Inoue and Yoshizo Matsuka : A double-layered complex cell sheet can regenerate the periodontal tissue in a mice model, Dentisphere, Surabayai, Nov. 2019.
2.
Arief Waskitho, Yumiko Yamamoto, Tsuyoshi Morita, Huijiao Yan, R Raju, Masamitsu Ohshima, Junhel Dalanon, Otto Baba and Yoshizo Matsuka : Bilateral effects of unilateral administration of botulinum toxin A in chemotherapy induced neuropathy, Dentisphere, Surabayai, Nov. 2019.
Waskitho Arief, Yamamoto Yumiko, Tsuyoshi Morita, Yan Huijiao, Raju Resmi, Masamitsu Ohshima, Dalanon Junhel, Otto Baba and Yoshizo Matsuka : Orofacial neuropathic pain rat models induced by chemotherapy drugs, Tokushima University Bioscience Retreat, Sep. 2019.
10.
Raju Resmi, Masamitsu Ohshima, Miho Inoue, Tsuyoshi Morita, Yan Huijiao, Otto Baba, Inoue Masahisa and Yoshizo Matsuka : Development of a multilayered complex cell sheet to regenerate periodontal tissue, Tokushima University Bioscience Retreat, Sep. 2019.
11.
Shin Matsumoto and Tsuyoshi Morita : Expression of mineralized tissue formation-related genes involved in morphogenesis of tooth and periodontal tissue, 令和元年度(第25回)スチューデント・クリニシャン・リサーチ・プログラム, Aug. 2019.