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Alveolar bone loss caused by periodontal disease eventually leads to tooth loss. Periodontal ligament stem cells (PDLSCs) are the tissue-specific cells for maintaining and repairing the periodontal ligament, cementum, and alveolar bone. Here, we investigated the role of erythropoietin receptor (EPOR), which regulates the microenvironment-modulating function of mesenchymal stem cells, in PDLSC-based periodontal therapy. We isolated PDLSCs from patients with chronic periodontal disease and healthy donors, referred to as PD-PDLSCs and Cont-PDLSCs, respectively. PD-PDLSCs exhibited reduced potency of periodontal tissue regeneration and lower expression of EPOR compared to Cont-PDLSCs. EPOR-silencing suppressed the potency of Cont-PDLSCs mimicking PD-PDLSCs, whereas EPO-mediated EPOR activation rejuvenated the reduced potency of PD-PDLSCs. Furthermore, we locally transplanted EPOR-silenced and EPOR-activated PDLSCs into the gingiva around the teeth of ligament-induced periodontitis model mice and demonstrated that EPOR in PDLSCs participated in the regeneration of the periodontal ligament, cementum, and alveolar bone in the ligated teeth. The EPOR-mediated paracrine function of PDLSCs maintains periodontal immune suppression and bone metabolic balance via osteoclasts and osteoblasts in the periodontitis model mice. Taken together, these results suggest that EPOR signaling is crucial for PDLSC-based periodontal regeneration and paves the way for the development of novel options for periodontal therapy.

DOI： 10.1038/s41598-024-57361-y

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The metastases of breast cancer to bone often cause osteolytic lesions not only by stimulating osteoclasts to resorb the bone but also by inhibiting osteoblasts from bone formation. Although tumor cell-derived extracellular vesicles (EVs) promote osteoclast differentiation and bone resorption, their roles in osteoblast differentiation and functions have not been elucidated. In this study, we investigated the effects of breast cancer cell-derived EVs on osteoblast differentiation and functions in vitro. We found that upon osteogenic induction, 4T1 bone metastatic mouse mammary tumor cell-derived EVs (4T1-EVs) were inhibited matrix mineralization of ST2 mouse bone marrow stromal cells. Temporal expression analysis of osteoblast marker genes, including runt-related transcription factor 2 (Runx2), osterix (Osx), alkaline phosphatase (Alp), collagen type I (Col1a1), bone sialoprotein (Bsp), and osteocalcin (Bglap) revealed that 4T1-EVs decreased their expression during the late stage of osteoblast differentiation. Elevated levels of c-Jun N-terminal kinase (JNK) phosphorylation, upon osteogenic induction, were diminished by 4T1-EVs, significantly. In contrast, the nullification of reduced JNK phosphorylation by anisomycin, a potent JNK activator, increased the expression levels of osteoblast differentiation markers. Overall, our data indicated that 4T1-EVs affect osteoblast maturation, at least partially, through the regulation of JNK activity, which provides novel insights into the pathological impact of osteolytic bone metastasis and the role of EVs in osteoblast differentiation.

DOI： 10.1016/j.abb.2023.109821

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Hirschsprung disease (HSCR) and its associated disorders (AD-HSCR) often result in severe hypoperistalsis caused by enteric neuropathy, mesenchymopathy, and myopathy. Notably, HSCR involving the small intestine, isolated hypoganglionosis, chronic idiopathic intestinal pseudo-obstruction, and megacystis-microcolon-intestinal hypoperistalsis syndrome carry a poor prognosis. Ultimately, small-bowel transplantation (SBTx) is necessary for refractory cases, but it is highly invasive and outcomes are less than optimal, despite advances in surgical techniques and management. Thus, regenerative therapy has come to light as a potential form of treatment involving regeneration of the enteric nervous system, mesenchyme, and smooth muscle in affected areas. We review the cutting-edge regenerative therapeutic approaches for managing HSCR and AD-HSCR, including the use of enteric nervous system progenitor cells, embryonic stem cells, induced pluripotent stem cells, and mesenchymal stem cells as cell sources, the recipient intestine's microenvironment, and transplantation methods. Perspectives on the future of these treatments are also discussed.

DOI： 10.1007/s00595-023-02741-6

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：The Japanese Society for Dental Materials and Devices  

<p>Carbonate apatite (CO₃Ap) is a major inorganic bone component and an effective bone substitute. To clarify the function of CO₃Ap, we compared differences among CO₃Ap, hydroxyapatite (HAp), and β-tricalcium phosphate (β-TCP) by focusing on mesenchymal stem cells (MSCs) that have a role in wound healing. For <i>in vivo</i> experiments, maxillary molars were removed and the bone substitute was inserted. MSC accumulation around extraction sockets was significantly promoted in CO₃Ap and β-TCP groups. For <i>in vitro</i> experiments, MSCs were cultured with bone substitutes. The differentiation potential and amount of calcium deposition were significantly lower in CO₃Ap and HAp groups than in the β-TCP group. Increases in insulin-like growth factor-I and vascular endothelial growth factor were found only in the CO₃Ap group. CO₃Ap-filled extraction sockets accumulated MSCs, and MSCs cultured in the presence of CO₃Ap produced large amounts of growth factors. These results suggest that CO₃Ap promotes healing of tooth extraction sockets.</p>

DOI： 10.4012/dmj.2022-040

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Systemic transplantation of mesenchymal stem cells (MSCs), such as bone marrow MSCs (BMMSCs) and stem cells from human exfoliated deciduous teeth (SHED), is considered a prominent treatment for osteopenia. However, the mechanism of action of the transplanted MSCs has been poorly elucidated. In the recipient target tissue, including bone and bone marrow, only a few donor MSCs can be detected, suggesting that the direct contribution of donor MSCs may not be expected for osteopenia treatment. Meanwhile, secretomes, especially contents within extracellular vesicles (EVs) released from donor MSCs (MSC-EVs), play key roles in the treatment of several diseases. In this context, administrated donor MSC-EVs may affect bone-forming function of recipient cells. In this review, we discuss how MSC-EVs contribute to bone recovery recipient tissue in osteopenia. We also summarize a novel mechanism of action of systemic administration of SHED-derived EVs (SHED-EVs) in osteopenia. We found that reduced telomerase activity in recipient BMMSCs caused the deficiency of microenvironmental modulating function, including bone and bone marrow-like niche formation and immunomodulation in estrogen-deficient osteopenia model mice. Systemic administration of SHED-EVs could exert therapeutic effects on bone reduction via recovering the telomerase activity, leading to the rejuvenation of the microenvironmental modulating function in recipient BMMSCs, as seen in systemic transplantation of SHED. RNase-preconditioned donor SHED-EVs diminished the therapeutic benefits of administrated SHED-EVs in the recipient osteopenia model mice. These facts suggest that MSC-EV therapy targets the recipient BMMSCs to rejuvenate the microenvironmental modulating function via telomerase activity, recovering bone density. We then introduce future challenges to develop the reproducible MSC-EV therapy in osteopenia.

DOI： 10.3389/fendo.2023.1151429

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Language：English   Publisher：(一社)日本歯科理工学会  

骨補填材としての炭酸アパタイト(炭酸Ap)をハイドロキシアパタイト(HAp)およびβリン酸三カルシウム(β-TCP)と比較し、間葉系幹細胞(MSC)に注目してそれらの間にみられる差異を検討した。6週齢の雄性ラット30匹を使用してin vivo実験を施行した。上顎の第1・2大臼歯を抜歯し、抜歯窩に上記3種の骨補填材のいずれかを移植した。その3日後と7日後に深麻酔死させ各種の評価に供した。その結果、炭酸Ap群とβ-TCP群では、抜歯窩付近のMSCの集積が有意に促進された。MSCを骨補填材の存在下で培養するin vitro実験も行ったところ、炭酸Ap群とHAp群では分化能とカルシウム沈着量がβ-TCP群よりも有意に低減していた。インスリン様成長因子I(IGF-I)と血管内皮細胞増殖因子(VEGF)が増加しているという結果は炭酸Ap群のみで認められた。炭酸Apを充填した抜歯窩にはMSCが集積し、炭酸Ap存在下で培養したMSCは成長因子類を大量に産生したことから、炭酸Apは抜歯窩の治癒を促進することが示唆された。

Language：English   Publisher：Journal of Cellular Physiology  

Nuclear protein 1 (NUPR1) is a stress-induced protein activated by various stresses, such as inflammation and oxidative stress. We previously reported that Nupr1 deficiency increased bone volume by enhancing bone formation in 11-week-old mice. Analysis of differentially expressed genes between wild-type (WT) and Nupr1-knockout (Nupr1-KO) osteocytes revealed that high temperature requirement A 1 (HTRA1), a serine protease implicated in osteogenesis and transforming growth factor-β signaling was markedly downregulated in Nupr1-KO osteocytes. Nupr1 deficiency also markedly reduced HtrA1 expression, but enhanced SMAD1 signaling in in vitro-cultured primary osteoblasts. In contrast, Nupr1 overexpression enhanced HtrA1 expression in osteoblasts, suggesting that Nupr1 regulates HtrA1 expression, thereby suppressing osteoblastogenesis. Since HtrA1 is also involved in cellular senescence and age-related diseases, we analyzed aging-related bone loss in Nupr1-KO mice. Significant spine trabecular bone loss was noted in WT male and female mice during 6−19 months of age, whereas aging-related trabecular bone loss was attenuated, especially in Nupr1-KO male mice. Moreover, cellular senescence-related markers were upregulated in the osteocytes of 6−19-month-old WT male mice but markedly downregulated in the osteocytes of 19-month-old Nupr1-KO male mice. Oxidative stress-induced cellular senescence stimulated Nupr1 and HtrA1 expression in in vitro-cultured primary osteoblasts, and Nupr1 overexpression enhanced p16ink4a expression in osteoblasts. Finally, NUPR1 expression in osteocytes isolated from the bones of patients with osteoarthritis was correlated with age. Collectively, these results indicate that Nupr1 regulates HtrA1-mediated osteoblast differentiation and senescence. Our findings unveil a novel Nupr1/HtrA1 axis, which may play pivotal roles in bone formation and age-related bone loss.

DOI： 10.1002/jcp.30949

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Objective: Chronic liver diseases often involve metabolic damage to the skeletal system. The underlying mechanism of bone loss in chronic liver diseases remains unclear, and appropriate therapeutic options, except for orthotopic liver transplantation, have proved insufficient for these patients. This study aimed to investigate the efficacy and mechanism of transplantation of immature hepatocyte-like cells converted from stem cells from human exfoliated deciduous teeth (SHED-Heps) in bone loss of chronic liver fibrosis. Methods: Mice that were chronically treated with CCl4 received SHED-Heps, and trabecular bone density, reactive oxygen species (ROS), and osteoclast activity were subsequently analyzed in vivo and in vitro. The effects of stanniocalcin 1 (STC1) knockdown in SHED-Heps were also evaluated in chronically CCl4 treated mice. Results: SHED-Hep transplantation (SHED-HepTx) improved trabecular bone loss and liver fibrosis in chronic CCl4-treated mice. SHED-HepTx reduced hepatic ROS production and interleukin 17 (Il-17) expression under chronic CCl4 damage. SHED-HepTx reduced the expression of both Il-17 and tumor necrosis factor receptor superfamily 11A (Tnfrsf11a) and ameliorated the imbalance of osteoclast and osteoblast activities in the bone marrow of CCl4-treated mice. Functional knockdown of STC1 in SHED-Heps attenuated the benefit of SHED-HepTx including anti-bone loss effect by suppressing osteoclast differentiation through TNFSF11–TNFRSF11A signaling and enhancing osteoblast differentiation in the bone marrow, as well as anti-fibrotic and anti-ROS effects in the CCl4-injured livers. Conclusions: These findings suggest that targeting hepatic ROS provides a novel approach to treat bone loss resulting from chronic liver diseases.

DOI： 10.1016/j.molmet.2022.101599

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Aberrant osteoclast formation and activation are the hallmarks of osteolytic metastasis. Extracellular vesicles (EVs), released from bone metastatic tumor cells, play a pivotal role in the progression of osteolytic lesions. However, the mechanisms through which tumor cell–derived EVs regulate osteoclast differentiation and function have not been fully elucidated. In this study, we found that 4T1 bone metastatic mouse mammary tumor cell–derived EVs (4T1-EVs) are taken up by mouse bone marrow macrophages to facilitate osteoclastogenesis. Furthermore, treatment of mature osteoclasts with 4T1-EVs promoted bone resorption, which was accompanied by enhanced survival of mature osteoclasts through the negative regulation of caspase-3. By comparing the miRNA content in 4T1-EVs with that in 67NR nonmetastatic mouse mammary tumor cell–derived EVs (67NR-EVs), miR-92a-3p was identified as one of the most enriched miRNAs in 4T1-EVs, and its transfer into mature osteoclasts significantly reduced apoptosis. Bioinformatic and Western blot analyses revealed that miR-92a-3p directly targeted phosphatase and tensin homolog (PTEN) in mature osteoclasts, resulting in increased levels of phospho-Akt. Our findings provide novel insights into the EV-mediated regulation of osteoclast survival through the transfer of miR-92a-3p, which enhances mature osteoclast survival via the Akt survival signaling pathway, thus promoting bone resorption.

DOI： 10.1111/cas.15557

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Language：English   Publisher：John Wiley & Sons Australia, Ltd  

骨転移乳癌細胞由来細胞外小胞(EV)が破骨細胞の分化や機能に及ぼす影響とそのメカニズムについて検討した。マウス乳癌細胞株4T1由来EV(4T1-EV)がマウス骨髄マクロファージに取り込まれ、破骨細胞形成を促進することを見出した。成熟破骨細胞を4T1-EVで処理すると、骨吸収が促進され、カスパーゼ3を負に制御して成熟破骨細胞の生存が促進された。4T1-EVと非転移性マウス乳癌細胞株67NR由来EV(67NR-EV)におけるmiRNAの内容を比較した。その結果、miR-92a-3pが4T1-EVに最も濃縮されたmiRNAの一つとして特定され、miR-92a-3pの成熟破骨細胞への移入によりアポトーシスが著しく抑制された。さらに、miR-92a-3pは成熟破骨細胞においてphosphatase and tensin homolog(PTEN)を直接標的とし、リン酸化Aktのレベルを増加させた。以上より、マウス乳癌細胞株4T1由来EVに存在するmiR-92a-3pは、Akt生存シグナル経路を介して成熟破骨細胞の生存を高め、骨吸収を促進させることが示された。

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Human dental pulp stem cell (hDPSCs)-based therapy is a feasible option for regenerative medicine, such as dental pulp regeneration. Here, we show the steps needed to colony-forming unit-fibroblasts (CFU-F)-based isolation, expansion, and cryopreservation of hDPSCs for manufacturing clinical-grade products under a xenogeneic-free/serum-free condition. We also demonstrate the characterization of hDPSCs by CFU-F, flow cytometric, and in vitro multipotent assays. For complete details on the use and execution of this protocol, please refer to Iwanaka et al. (2020).

DOI： 10.1016/j.xpro.2022.101386

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Hirschsprung's disease is a congenital entero-neuropathy that causes chronic constipation and intestinal obstruction. New treatments for entero-neuropathy are needed because current surgical strategies have limitations5. Entero-neuropathy results from enteric nervous system dysfunction due to incomplete colonization of the distal intestine by neural crest-derived cells. Impaired cooperation between the enteric nervous system and intestinal pacemaker cells may also contribute to entero-neuropathy. Stem cell therapy to repair these multiple defects represents a novel treatment approach. Dental pulp stem cells derived from deciduous teeth (dDPSCs) are multipotent cranial neural crest-derived cells, but it remains unknown whether dDPSCs have potential as a new therapy for entero-neuropathy. Here we show that intravenous transplantation of dDPSCs into the Japanese Fancy-1 mouse, an established model of hypoganglionosis and entero-neuropathy, improves large intestinal structure and function and prolongs survival. Intravenously injected dDPSCs migrate to affected regions of the intestine through interactions between stromal cell-derived factor-1α and C-X-C chemokine receptor type-4. Transplanted dDPSCs differentiate into both pacemaker cells and enteric neurons in the proximal colon to improve electrical and peristaltic activity, in addition to their paracrine effects. Our findings indicate that transplanted dDPSCs can differentiate into different cell types to correct entero-neuropathy-associated defects.

DOI： 10.1038/s41598-022-10077-3

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Recent advances in mesenchymal stem/stromal cell (MSC) research have led us to consider the feasibility of MSC-based therapy for various diseases. Human dental pulp-derived MSCs (hDPSCs) have been identified in the dental pulp tissue of deciduous and permanent teeth, and they exhibit properties with self-renewal and in vitro multipotency. Interestingly, hDPSCs exhibit superior immunosuppressive functions toward immune cells, especially T lymphocytes, both in vitro and in vivo. Recently, hDPSCs have been shown to have potent immunomodulatory functions in treating systemic lupus erythematosus (SLE) in the SLE MRL/lpr mouse model. However, the mechanisms underlying the immunosuppressive efficacy of hDPSCs remain unknown. This review aims to introduce a new target of hDPSC-based therapy on the recipient niche function in SLE.

DOI： 10.3390/ijms23073479

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DOI： 10.1038/s41374-021-00656-9

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DOI： 10.1038/s41374-021-00661-y

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DOI： 10.1038/s41374-021-00661-y

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DOI： 10.1186/s13287-021-02652-8

Repository Public URL： https://hdl.handle.net/2324/7172586

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DOI： https://doi.org/10.1371/journal.pone.0259966

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DOI： 10.1038/s41374-021-00633-2

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DOI： 10.1038/s41374-021-00633-2

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DOI： 10.4049/jimmunol.2001312

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DOI： 10.1186/s13287-020-02113-8

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DOI： 10.1186/s13287-020-01818-0

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DOI： 10.1186/s13287-020-01630-w

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DOI： 10.1007/s00383-019-04564-4

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DOI： 10.1007/s00383-019-04564-4

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Wilson’s disease (WD) is an inherited metabolic disease arising from ATPase copper transporting beta gene (ATP7B) mutation. Orthotoropic liver transplantation is the only radical treatment of fulminant WD, although appropriate donors are lacking at the onset of emergency. Given the hepatogenic capacity and tissue-integration/reconstruction ability in the liver of stem cells from human exfoliated deciduous teeth (SHED), SHED have been proposed as a source for curing liver diseases. We hypothesized the therapeutic potential of SHED and SHED-converted hepatocyte-like- cells (SHED-Heps) for fulminant WD. SHED and SHED-Heps were transplanted into WD model Atp7b-mutated Long-Evans Cinnamon (LEC) rats received copper overloading to induce a lethal fulminant liver failure. Due to the superior copper tolerance via ATP7B, SHED-Hep transplantation gave more prolonged life-span of fulminant LEC rats than SHED transplantation. The integrated ATP7B-expressing SHED-Heps showed more therapeutic effects on to restoring the hepatic dysfunction and tissue damages in the recipient liver than the integrated naïve SHED without ATP7B expression. Moreover, SHED-Heps could reduce copper-induced oxidative stress via ATP7B- independent stanniocalcin 1 secretion in the fulminant LEC rats, suggesting a possible role for paracrine effect of the integrated SHED-Heps. Taken together, SHED-Heps offer a potential of functional restoring, bridging, and preventive approaches for treating fulminant WD.

DOI： 10.1038/s41598-018-38275-y

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DOI： 10.3390/app9224908

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DOI： 10.3390/app9224908

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DOI： 10.1038/s41374-018-0179-4

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DOI： 10.1038/s41374-018-0179-4

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DOI： 10.1016/j.joen.2019.01.016

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DOI： 10.1016/j.joen.2019.01.016

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DOI： 10.1007/s00595-019-01783-z

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Regenerative medicine using stem cells from human exfoliated deciduous teeth (SHED): a promising new treatment in pediatric surgery.
Stem cells from human exfoliated deciduous teeth (SHEDs), being a type of mesenchymal stem cell, are an ideal cell source for regenerative medicine. They have minimal risk of oncogenesis, high proliferative capacity, high multipotency, and immunosuppressive ability. Stem cell transplantation using SHED has been found to have an anti-fibrotic effect on liver fibrosis in mice. SHED transplantation and the bio 3D printer, which can create scaffold-free 3-D images of the liver and diaphragm, provide a new innovative treatment modality for intractable pediatric surgical diseases such as biliary atresia and diaphragmatic hernia.

DOI： 10.1007/s00595-019-01783-z

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DOI： 10.1038/s41598-018-21183-6

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DOI： 10.1186/s13287-018-1077-9

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DOI： 10.1186/s13287-018-1042-7

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Pamidronate decreases bilirubin-impaired cell death and improves dentinogenic dysfunction of stem cells from human deciduous teeth.
BACKGROUND: Hyperbilirubinemia that occurs in pediatric liver diseases such as biliary atresia can result in the development of not only jaundice in the brain, eyes, and skin, but also tooth abnormalities including green pigmentation and dentin hypoplasia in the developing teeth. However, hyperbilirubinemia-induced tooth impairments remain after liver transplantation. No effective dental management to prevent hyperbilirubinemia-induced tooth impairments has been established. METHODS: In this study, we focused on pamidronate, which is used to treat pediatric osteopenia, and investigated its effects on hyperbilirubinemia-induced tooth impairments. We cultured stem cells from human exfoliated deciduous teeth (SHED) under high and low concentrations of unconjugated bilirubin in the presence or absence of pamidronate. We then analyzed the effects of pamidronate on the cell death, associated signal pathways, and dentinogenic function in SHED. RESULTS: We demonstrated that a high concentration of unconjugated bilirubin induced cell death in SHED via the mitochondrial pathway, and this was associated with the suppression of AKT and extracellular signal-related kinase 1 and 2 (ERK1/2) signal pathways and activation of the nuclear factor kappa B (NF-κB) signal pathway. The high concentration of unconjugated bilirubin impaired the in vitro and in vivo dentinogenic capacity of SHED, but not the low concentration. We then demonstrated that pamidronate decreased the bilirubin-induced cell death in SHED via the altered AKT, ERK1/2, and NF-κB signal pathways and recovered the bilirubin-impaired dentinogenic function of SHED. CONCLUSIONS: Our findings suggest that pamidronate may prevent tooth abnormalities in pediatric patients with hyperbilirubinemia.

DOI： 10.1186/s13287-018-1042-7

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Suppression of AKT-mTOR signal pathway enhances osteogenic/dentinogenic capacity of stem cells from apical papilla.
BACKGROUND: Stem cells from apical papilla (SCAP) are a subpopulation of mesenchymal stem cells (MSCs) isolated from the apical papilla of the developing tooth root apex of human teeth. Because of their osteogenic/dentinogenic capacity, SCAP are considered as a source for bone and dentin regeneration. However, little is understood about the molecular mechanism of osteogenic/dentinogenic differentiation of SCAP. Phosphoinositide 3 kinase (PI3K)-AKT-mammalian target of rapamycin (mTOR) signal pathway participates in regulating the differentiation of various cell types, such as MSCs. In this study, we examined the role of the PI3K-AKT-mTOR signal pathway in the osteogenic/dentinogenic differentiation of SCAP. Moreover, we challenge to fabricate scaffold-free SCAP-based spheroidal calcified constructs. METHODS: SCAP were pretreated with or without small interfering RNA for AKT (AKT siRNA), PI3K inhibitor LY294402, and mTOR inhibitor rapamycin and were cultured under osteogenic/dentinogenic differentiation to examine in vitro and in vivo calcified tissue formation. Moreover, SCAP-based cell aggregates were pretreated with or without LY294402 and rapamycin. The cell aggregates were cultured under osteogenic/dentinogenic condition and were analyzed the calcification of the aggregates. RESULTS: Pretreatment with AKT siRNA, LY294402, and rapamycin enhances the in vitro and in vivo calcified tissue-forming capacity of SCAP. SCAP were fabricated as scaffold-free spheroids and were induced into forming calcified 3D constructs. The calcified density of the spheroidal constructs was enhanced when the spheroids were pretreated with LY294402 and rapamycin. CONCLUSIONS: Our findings indicate that the suppression of PI3K-AKT-mTOR signal pathway plays a role in not only enhancing the in vivo and in vitro osteogenic/dentinogenic differentiation of SCAP, but also promoting the calcification of scaffold-free SCAP-based calcified constructs. These findings suggest that a suppressive regulation of PI3K-AKT-mTOR signal pathway is a novel approach for SCAP-based bone and dentin regeneration.

DOI： 10.1186/s13287-018-1077-9

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Bilirubin reversibly affects cell death and odontogenic capacity in stem cells from human exfoliated deciduous teeth.

DOI： 10.1111/odi.12827

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DOI： 10.1111/odi.12827

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DOI： 10.33425/2639-9490.1018

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Exogenous nitric oxide stimulates the odontogenic differentiation of rat dental pulp stem cells.
Nitric oxide (NO) is thought to play a pivotal regulatory role in dental pulp tissues under both physiological and pathological conditions. However, little is known about the NO functions in dental pulp stem cells (DPSCs). We examined the direct actions of a spontaneous NO gas-releasing donor, NOC-18, on the odontogenic capacity of rat DPSCs (rDPSCs). In the presence of NOC-18, rDPSCs were transformed into odontoblast-like cells with long cytoplasmic processes and a polarized nucleus. NOC-18 treatment increased alkaline phosphatase activity and enhanced dentin-like mineralized tissue formation and the expression levels of several odontoblast-specific genes, such as runt related factor 2, dentin matrix protein 1 and dentin sialophosphoprotein, in rDPSCs. In contrast, carboxy-PTIO, a NO scavenger, completely suppressed the odontogenic capacity of rDPSCs. This NO-promoted odontogenic differentiation was activated by tumor necrosis factor-NF-κB axis in rDPSCs. Further in vivo study demonstrated that NOC-18-application in a tooth cavity accelerated tertiary dentin formation, which was associated with early nitrotyrosine expression in the dental pulp tissues beneath the cavity. Taken together, the present findings indicate that exogenous NO directly induces the odontogenic capacity of rDPSCs, suggesting that NO donors might offer a novel host DPSC-targeting alternative to current pulp capping agents in endodontics.

DOI： 10.1038/s41598-018-21183-6

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As osteoclasts have the central roles in normal bone remodeling, it is ideal to regulate only the osteoclasts performing pathological bone destruction without affecting normal osteoclasts. Based on a hypothesis that pathological osteoclasts form under the pathological microenvironment of the bone tissues, we here set up optimum culture conditions to examine the entity of pathologically activated osteoclasts (PAOCs). Through searching various inflammatory cytokines and their combinations, we found the highest resorbing activity of osteoclasts when osteoclasts were formed in the presence of M-CSF, receptor activator of NF-κB ligand, and IL-1β. We have postulated that these osteoclasts are PAOCs. Analysis using confocal laser microscopy revealed that PAOCs showed extremely high proton secretion detected by the acid-sensitive fluorescence probe Rh-PM and bone resorption activity compared with normal osteoclasts. PAOCs showed unique morphology bearing high thickness and high motility with motile cellular processes in comparison with normal osteoclasts. We further examined the expression of Kindlin-3 and Talin-1, essential molecules for activating integrin b-chains. Although normal osteoclasts express high levels of Kindlin-3 and Talin-1, expression of these molecules was markedly suppressed in PAOCs, suggesting the abnormality in the adhesion property. When whole membrane surface of mature osteoclasts was biotinylated and analyzed, the IL-1β-induced cell surface protein was detected. PAOCs could form a subpopulation of osteo-clasts possibly different from normal osteoclasts. PAOC-specific molecules could be an ideal target for regulating pathological bone destruction.

DOI： 10.4049/jimmunol.1602035

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DOI： 10.4049/jimmunol.1602035

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1186/s40729-017-0112-4

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1038/s41598-017-14542-2

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1038/labinvest.2017.55

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1038/labinvest.2017.55

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1038/s41598-017-14542-2

Language：Japanese  

Language：Japanese  

Language：Japanese  

Language：English   Publishing type：Research paper (scientific journal)  

Therapeutic interactions between mesenchymal stem cells for healing medication-related osteonecrosis of the jaw.
BACKGROUND: Mesenchymal stem cells (MSCs) have been isolated from a variety of tissues, including bone marrow, adipose, and mucosa. MSCs have the capacity for self-renewal and differentiation. Reports have been published on the systemic administration of MSCs leading to functional improvements by engraftment and differentiation, thus providing a new strategy to regenerate damaged tissues. Recently, it has become clear that MSCs possess immunomodulatory properties and can therefore be used to treat diseases. However, the therapeutic effect mechanisms of MSCs are yet to be determined. Here, we investigated these mechanisms using a medication-related osteonecrosis of the jaw (MRONJ)-like mouse model. METHODS: To generate MRONJ-like characteristics, mice received intravenous zoledronate and dexamethasone two times a week. At 1 week after intravenous injection, maxillary first molars were extracted, and at 1 week after tooth extraction, MSCs were isolated from the bone marrow of the mice femurs and tibias. To compare "diseased MSCs" from MRONJ-like mice (d-MSCs) with "control MSCs" from untreated mice (c-MSCs), the isolated MSCs were analyzed by differentiation and colony-forming unit-fibroblast (CFU-F) assays and systemic transplantation of either d-MSCs or c-MSCs into MRONJ-like mice. Furthermore, we observed the exchange of cell contents among d-MSCs and c-MSCs during coculture with all combinations of each MSC type. RESULTS: d-MSCs were inferior to c-MSCs in differentiation and CFU-F assays. Moreover, the d-MSC-treated group did not show earlier healing in MRONJ-like mice. In cocultures with any combination, MSC pairs formed cell-cell contacts and exchanged cell contents. Interestingly, the exchange among c-MSCs and d-MSCs was more frequently observed than other pairs, and d-MSCs were distinguishable from c-MSCs. CONCLUSIONS: The interaction of c-MSCs and d-MSCs, including exchange of cell contents, contributes to the treatment potential of d-MSCs. This cellular behavior might be one therapeutic mechanism used by MSCs for MRONJ.

DOI： 10.1186/s13287-016-0367-3

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1186/s13287-016-0367-3

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1038/srep19286

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1038/srep19286

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1186/s13287-015-0154-6

Language：English   Publishing type：Research paper (scientific journal)  

In vivo hepatogenic capacity and therapeutic potential of stem cells from human exfoliated deciduous teeth in liver fibrosis in mice.
INTRODUCTION: Liver transplantation is a gold standard treatment for intractable liver diseases. Because of the shortage of donor organs, alternative therapies have been required. Due to their potential to differentiate into a variety of mature cells, stem cells are considered feasible cell sources for liver regeneration. Stem cells from human exfoliated deciduous teeth (SHED) exhibit hepatogenic capability in vitro. In this study, we investigated their in vivo capabilities of homing and hepatocyte differentiation and therapeutic efficacy for liver disorders in carbon tetrachloride (CCl4)-induced liver fibrosis model mice. METHODS: We transplanted SHED into CCl4-induced liver fibrosis model mice through the spleen, and analyzed the in vivo homing and therapeutic effects by optical, biochemical, histological, immunological and molecular biological assays. We then sorted human leukocyte antigen-ABC (HLA-ABC)-positive cells from primary CCl4-damaged recipient livers, and analyzed their fusogenicity and hepatic characteristics by flow cytometric, genomic DNA, hepatocyte-specific gene assays. Furthermore, we examined the treatment effects of HLA-positive cells to a hepatic dysfunction by a secondary transplantation into CCl4-treated mice. RESULTS: Transplanted SHED homed to recipient livers, and expressed HLA-ABC, human hepatocyte specific antigen hepatocyte paraffin 1 and human albumin. SHED transplantation markedly recovered liver dysfunction and led to anti-fibrotic and anti-inflammatory effects in the recipient livers. SHED-derived HLA-ABC-positive cells that were sorted from the primary recipient liver tissues with CCl4 damage did not fuse with the host mouse liver cells. Sorted HLA-positive cells not only expressed human hepatocyte-specific genes including albumin, cytochrome P450 1A1, fumarylacetoacetase, tyrosine aminotransferase, uridine 5'-diphospho-glucuronosyltransferase, transferrin and transthyretin, but also secreted human albumin, urea and blood urea nitrogen. Furthermore, SHED-derived HLA-ABC-positive cells were secondary transplanted into CCl4-treated mice. The donor cells homed into secondary recipient livers, and expressed hepatocyte paraffin 1 and human albumin, as well as HLA-ABC. The secondary transplantation recovered a liver dysfunction in secondary recipients. CONCLUSIONS: This study indicates that transplanted SHED improve hepatic dysfunction and directly transform into hepatocytes without cell fusion in CCl4-treated mice, suggesting that SHED may provide a feasible cell source for liver regeneration.

DOI： 10.1186/s13287-015-0154-6

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.stem.2015.04.016

Language：English   Publishing type：Research paper (scientific journal)  

Transplantation of mesenchymal stem cells ameliorates secondary osteoporosis through interleukin-17-impaired functions of recipient bone marrow mesenchymal stem cells in MRL/lpr mice.
INTRODUCTION: Secondary osteoporosis is common in systemic lupus erythematosus and leads to a reduction in quality of life due to fragility fractures, even in patients with improvement of the primary disorder. Systemic transplantation of mesenchymal stem cells could ameliorate bone loss and autoimmune disorders in a MRL/lpr mouse systemic lupus erythematosus model, but the detailed therapeutic mechanism of bone regeneration is not fully understood. In this study, we transplanted human bone marrow mesenchymal stem cells (BMMSCs) and stem cells from exfoliated deciduous teeth (SHED) into MRL/lpr mice and explored their therapeutic mechanisms in secondary osteoporotic disorders of the systemic lupus erythematosus model mice. METHODS: The effects of systemic human mesenchymal stem cell transplantation on bone loss of MRL/lpr mice were analyzed in vivo and ex vivo. After systemic human mesenchymal stem cell transplantation, recipient BMMSC functions of MRL/lpr mice were assessed for aspects of stemness, osteogenesis and osteoclastogenesis, and a series of co-culture experiments under osteogenic or osteoclastogenic inductions were performed to examine the efficacy of interleukin (IL)-17-impaired recipient BMMSCs in the bone marrow of MRL/lpr mice. RESULTS: Systemic transplantation of human BMMSCs and SHED recovered the reduction in bone density and structure in MRL/lpr mice. To explore the mechanism, we found that impaired recipient BMMSCs mediated the negative bone metabolic turnover by enhanced osteoclastogenesis and suppressed osteoblastogenesis in secondary osteoporosis of MRL/lpr mice. Moreover, IL-17-dependent hyperimmune conditions in the recipient bone marrow of MRL/lpr mice damaged recipient BMMSCs to suppress osteoblast capacity and accelerate osteoclast induction. To overcome the abnormal bone metabolism, systemic transplantation of human BMMSCs and SHED into MRL/lpr mice improved the functionally impaired recipient BMMSCs through IL-17 suppression in the recipient bone marrow and then maintained a regular positive bone metabolism via the balance of osteoblasts and osteoclasts. CONCLUSIONS: These findings indicate that IL-17 and recipient BMMSCs might be a therapeutic target for secondary osteoporosis in systemic lupus erythematosus.

DOI： 10.1186/s13287-015-0091-4

Language：English  

DOI： 10.1016/j.stem.2015.04.016

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1186/s13287-015-0091-4

Language：Japanese  

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1007/s00383-014-3576-9

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1007/s00383-014-3576-9

Language：Japanese  

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1038/labinvest.2013.152

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1002/emmm.201303000

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1038/labinvest.2013.152

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1371/journal.pone.0090681

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1002/emmm.201303000

Language：English   Publishing type：Research paper (scientific journal)  

Abnormal stem cell function makes a known contribution to many malignant tumors, but the role of stem cells in benign tumors is not well understood. Here, we show that ossifying fibroma (OF) contains a stem cell population that resembles mesenchymal stem cells (OFMSCs) and is capable of generating OF-like tumor xenografts. Mechanistically, OFMSCs show enhanced TGF-β signaling that induces aberrant proliferation and deficient osteogenesis via Notch and BMP signaling pathways, respectively. The elevated TGF-β activity is tightly regulated by JHDM1D-mediated epigenetic regulation of thrombospondin-1 (TSP1), forming a JHDM1D/TSP1/TGF-β/SMAD3 autocrine loop. Inhibition of TGF-β signaling in OFMSCs can rescue their abnormal osteogenic differentiation and elevated proliferation rate. Furthermore, chronic activation of TGF-β can convert normal MSCs into OF-like MSCs via establishment of this JHDM1D/TSP1/TGF-β/SMAD3 autocrine loop. These results reveal that epigenetic regulation of TGF-β signaling in MSCs governs the benign tumor phenotype in OF and highlight TGF-β signaling as a candidate therapeutic target.

DOI： 10.1016/j.stem.2013.08.010

Language：English   Publishing type：Research paper (scientific journal)  

Abnormal stem cell function makes a known contribution to many malignant tumors, but the role of stem cells in benign tumors is not well understood. Here, we show that ossifying fibroma (OF) contains a stem cell population that resembles mesenchymal stem cells (OFMSCs) and is capable of generating OF-like tumor xenografts. Mechanistically, OFMSCs show enhanced TGF-β signaling that induces aberrant proliferation and deficient osteogenesis via Notch and BMP signaling pathways, respectively. The elevated TGF-β activity is tightly regulated by JHDM1D-mediated epigenetic regulation of thrombospondin-1 (TSP1), forming a JHDM1D/TSP1/TGF-β/SMAD3 autocrine loop. Inhibition of TGF-β signaling in OFMSCs can rescue their abnormal osteogenic differentiation and elevated proliferation rate. Furthermore, chronic activation of TGF-β can convert normal MSCs into OF-like MSCs via establishment of this JHDM1D/TSP1/TGF-β/SMAD3 autocrine loop. These results reveal that epigenetic regulation of TGF-β signaling in MSCs governs the benign tumor phenotype in OF and highlight TGF-β signaling as a candidate therapeutic target.

Language：English   Publishing type：Research paper (scientific journal)  

The role of phosphoinositide 3-kinase in adhesion of oral epithelial cells to titanium.
BACKGROUND: Oral epithelial cells (OECs) adhesion to titanium may improve the success rate of implant restoration. PURPOSE: We investigated the mechanism by which OECs adhere to titanium dental implants. MATERIALS AND METHODS: (1) After culturing rat OECs on titanium plates (Ti) or culture dishes in the presence or absence of a phosphoinositide 3-kinase (PI3K) activator or inhibitors and/or growth factors, and OEC morphology under these conditions were analyzed. (2) Right maxillary first molars were extracted and replaced with experimental implants. The rats were treated with or without growth factors. RESULTS: (1) Cell adherence was lower of OECs on Ti than in those on culture dishes, as were the levels of integrin β4 and the continuity of F-actin structures. After PI3K inhibition, markedly reducing adherence to both substrates. In contrast, PI3K activation with activator or insulin-like growth factor restored the OEC adherence and the expression of adhesion molecules on Ti to the levels seen in OECs cultured on dishes. Cell migration was inhibited by PI3K activation. (2) High expression of integrin β4 was observed in the peri-implant epithelia of PI3K-activated rats. CONCLUSION: These findings suggest that PI3K plays an important role in the adhesion of OECs to Ti.

DOI： 10.1016/j.archoralbio.2013.07.013

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1002/jbm.a.34608

Language：English   Publishing type：Research paper (scientific journal)  

Promotive effect of insulin-like growth factor-1 for epithelial sealing to titanium implants
Improvement of oral epithelial adhesion to titanium (Ti) may significantly enhance the efficacy of dental implants. Here, we investigated whether insulin-like growth factor-1 (IGF-1) improved the sealing of the peri-implant epithelium (PIE) around the implant. Right maxillary first molars were extracted from rats and replaced with experimental implants. After 4 weeks of IGF-1 treatment, the implant-PIE interface exhibited a band of immunoreactive laminin-332 (Ln-5), similar to the tooth-junctional epithelium interface, that was partially absent in the untreated group. Immunoelectron microscopy showed a characteristic Ln-5-positive band including hemidesmosomes at both the apical and upper portions of the implant-PIE interface in the IGF-1-treated group. We also investigated the effects of IGF-1/PI3K inhibitors on the dynamics of rat oral epithelial cells (OECs) grown on Ti plates. In OECs cultured with IGF-1, adhesion protein expression increased, cell adherence to Ti plates was higher, and proliferation was faster, whereas migration and apoptosis were induced in the absence of IGF-1 or in the presence of both IGF-1 and a PI3K inhibitor. These data suggest that PI3K mediates the promotive effects of IGF-1, and that IGF-1 is effective at enhancing epithelial integration around Ti implants.

DOI： 10.1002/jbm.a.34608

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.archoralbio.2013.07.013

Language：Japanese  

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1177/0022034513488787

Language：Others  

Immune therapeutic potential of stem cells from human supernumerary teeth.

DOI： 10.1177/0022034513490732

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1177/0022034513490732

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1002/jcb.24433

Language：English   Publishing type：Research paper (scientific journal)  

Tunneling nanotube formation is essential for the regulation of osteoclastogenesis.
Osteoclasts are the multinucleated giant cells formed by cell fusion of mononuclear osteoclast precursors. Despite the finding of several membrane proteins involving DC-STAMP as regulatory proteins required for fusion among osteoclast precursors, cellular and molecular events concerning this process are still ambiguous. Here we identified Tunneling Nanotubes (TNTs), long intercellular bridges with small diameters, as the essential cellular structure for intercellular communication among osteoclast precursors in prior to cell fusion. Formation of TNTs was highly associated with osteoclastogenesis and it was accompanied with the significant induction of the M-Sec gene, an essential gene for TNT formation. M-Sec gene expression was significantly upregulated by RANKL-treatment in osteoclast precursor cell line. Blockage of TNT formation by Latrunclin B or by M-Sec siRNA significantly suppressed osteoclastogenesis. We have detected the rapid intercellular transport of not only the membrane phospholipids labeled with DiI but also the DC-STAMP-GFP fusion protein through TNTs formed among osteoclast precursors during osteoclastogenesis. Transportation of such regulatory molecules through TNTs would be essential for the process of the specific cell fusion among osteoclast precursors.

DOI： 10.1002/jcb.24433

Language：English   Publishing type：Research paper (other academic)  

DOI： 10.4028/www.scientific.net/KEM.529-530.407

Language：English  

Expression of Integrin alpha-3 and beta-4 subunits on the process of peri-implant epithelium formation
Integrin, a component of the hemidesmosome, plays a role for epithelial cell migration and adhesion. This study investigated the process of peri-implant epithelium (PIE) formation after implantation, and compared it to the process of oral mucosa healing after tooth extraction. At the healing site of extraction socket without implant, the original junctional epithelium (JE) had disappeared at week 2, and the oral epithelium (OE) with integrin-α3 positive basal cells extending from the sides of the wound, then joined in the middle of the extraction socket. On the other hand in implant group, newly formed epithelium with integrin-α3 positive cells from the OE extended apically 1 week after implantation. After 3 weeks, basal cells of the new epithelium consisted of those with integrin-α3 positive but β4 negative. Finally, after 4 weeks, integrin-β4 was expressed at the implant-PIE interface. These findings suggest that integrin α3β1 plays a role in cell migration during PIE formation from OE. Furthermore, after the completion of PIE constitution, integrin α6β4 contributes to the attachment to titanium. © (2013) Trans Tech Publications, Switzerland.

DOI： 10.4028/www.scientific.net/KEM.529-530.407

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1186/scrt131

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1371/journal.pone.0051777

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1371/journal.pone.0051777

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1155/2012/273291

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.stem.2012.03.007

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.stem.2012.03.007

Language：Japanese  

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1177/0022034510387796

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1177/0022034510387796

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1186/scrt5

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1242/dev.051672

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1242/dev.051672

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1002/jbmr.37

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.2217/rme.10.30

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.2217/rme.10.30

Language：English   Publishing type：Research paper (scientific journal)  

Skeletal muscles are formed from two cell lineages, myogenic and fibroblastic. Mesoderm-derived myogenic progenitors form muscle cells whereas fibroblastic cells give rise to the supportive connective tissue of skeletal muscles, such as the tendons and perimysium. It remains unknown how myogenic and fibroblastic cell-cell interactions affect cell fate determination and the organization of skeletal muscle. In the present study, we investigated the functional significance of cell-cell interactions in regulating skeletal muscle development. Our study shows that cranial neural crest (CNC) cells give rise to the fibroblastic cells of the tongue skeletal muscle in mice. Loss of Tgfbr2 in CNC cells (Wnt1-Cre;Tgfbr2^flox/flox) results in microglossia with reduced Scleraxis and Fgf10 expression as well as decreased myogenic cell proliferation, reduced cell number and disorganized tongue muscles. Furthermore, TGF-β2 beads induced the expression of Scleraxis in tongue explant cultures. The addition of FGF10 rescued the muscle cell number in Wnt1-Cre;Tgfbr2^flox/flox mice. Thus, TGF-β induced FGF10 signaling has a critical function in regulating tissue-tissue interaction during tongue skeletal muscle development.

DOI： 10.1016/j.ydbio.2010.02.030

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.ydbio.2010.02.030

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1089/ten.tea.2009.0518

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1089/ten.TEA.2009.0518

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1111/j.1601-0825.2009.01593.x

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1371/journal.pone.0007798

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1371/journal.pone.0007798

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1007/s00418-009-0616-y

Language：English   Publishing type：Research paper (scientific journal)  

TRPV2 expression in rat oral mucosa.
The oral mucosa is a highly specialised, stratified epithelium that confers protection from infection and physical, chemical and thermal stimuli. The non-keratinised junctional epithelium surrounds each tooth like a collar and is easily attacked by foreign substances from the oral sulcus. We found that TRPV2, a temperature-gated channel, is highly expressed in junctional epithelial cells, but not in oral sulcular epithelial cells or oral epithelial cells. Dual or triple immunolabelling with immunocompetent cell markers also revealed TRPV2 expression in Langerhans cells and in dendritic cells and macrophages. Electron microscopy disclosed TRPV2 immunoreactivity in the unmyelinated and thinly myelinated axons within the connective tissue underlying the epithelium. TRPV2 labelling was also observed in venule endothelial cells. The electron-dense immunoreaction in junctional epithelial cells, macrophages and neural axons occurred on the plasma membrane, on invaginations of the plasma membrane and in vesicular structures. Because TRPV2 has been shown to respond to temperature, hypotonicity and mechanical stimuli, gingival cells expressing TRPV2 may act as sensor cells, detecting changes in the physical and chemical environment, and may play a role in subsequent defence mechanisms.

DOI： 10.1007/s00418-009-0616-y

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1038/ncb1913

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1038/ncb1913

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1002/stem.68

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1002/stem.68

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1111/j.1601-0825.2009.01564.x

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1002/stem.2

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1002/stem.2

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1182/blood-2008-10-182246

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1182/blood-2008-10-182246

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1007/s00441-008-0720-7

Language：English   Publishing type：Research paper (scientific journal)  

Distribution of substance P and neurokinin-1 receptors in the peri-implant epithelium around titanium dental implants in rats.
We examined the distribution of substance P and neurokinin-1 (NK1) receptors and substance-P-containing nerve fibers in the peri-implant mucosa around titanium dental implants in rats. Immunohistochemistry and immunocytochemistry revealed that substance-P-immunoreactive nerve fibers abundantly innervated the peri-implant epithelium (PIE) compared with other epithelia of the peri-implant mucosa. NK1 receptor mRNA and protein expression in the peri-implant mucosa were confirmed by reverse transcription with the polymerase chain reaction and immunoblotting. Immunoelectron microscopy revealed that NK1 receptor immunoreactivity was preferentially localized in peri-implant epithelial cells. NK1-receptor-positive products were found on the plasma membrane and in vesicles and granules in PIE cells. Neutrophils and intraepithelial nerve axons in the PIE were positive for the NK1 receptor. NK1 receptor immunoreactivity was also detected in endothelial cells, fibroblasts, and nerve fibers in the connective tissue beneath the PIE. These findings suggest that peri-implant tissue receives sensory information through regenerated nerves expressing substance P and the NK1 receptor. In the peri-implant mucosa, the substance P/NK1 receptor system may play a role in pain transmission, the endocytosis of neutrophils, the extravasation of crevicular fluid, and the migration of macrophages and neutrophils in response to neurogenic inflammation, as in healthy gingiva.

DOI： 10.1007/s00441-008-0720-7

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.2217/17460816.3.6.499

Language：English   Publishing type：Research paper (scientific journal)  

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DOI： 10.1371/journal.pone.0002615

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DOI： 10.1111/j.1601-0825.2007.01396.x

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1371/journal.pone.0002615

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1111/j.1601-0825.2007.01396.x

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.joen.2007.11.021

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.joen.2007.11.021

Language：Japanese  

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.bbrc.2007.06.028

Language：English   Publishing type：Research paper (scientific journal)  

Cystatin C stimulates the differentiation of mouse osteoblastic cells and bone formation.
Cystatin C (CysC) is a natural cysteine proteinase inhibitor that suppresses the differentiation and bone-resorptive function of osteoclasts. By contrast, the effect of CysC on the differentiation and bone-formative function of osteoblasts has not been elucidated thoroughly. We examined the effects of CysC on mouse osteoblastic cells using in vitro cultures from bone marrow and calvaria and ex vivo calvarial cultures. CysC-stimulated cells showed increased alkaline phosphatase (ALP) activity, mineralization of the new bone matrix, and calvarial bone formation. The cells treated with CysC immunodepleted by anti-CysC antibody (iCysC) and a chemical papain-like cysteine proteinase inhibitor, E-64, did not induce mineralization. Elevated mRNA levels of bone morphogenetic protein (BMP)-2, the differentiation marker osteocalcin, and a master osteogenic transcription factor, Runx2, were observed in CysC-treated cells. These results suggest that CysC affects the BMP signaling cascades in osteoblastic cells and then promotes osteoblast differentiation, mineralization, and bone formation.

DOI： 10.1016/j.bbrc.2007.06.028

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1177/154405910708600703

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1177/154405910708600703

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1007/s00441-005-0003-5

Language：English   Publishing type：Research paper (scientific journal)  

Language：English   Publishing type：Research paper (scientific journal)  

Sequential expression of endothelial nitric oxide synthase, inducible nitric oxide synthase, and nitrotyrosine in odontoblasts and pulp cells during dentin repair after tooth preparation in rat molars.
Nitric oxide (NO) stimulates osteoblast differentiation, but whether NO contributes to odontoblast differentiation during dentin repair is unknown. By using reverse transcription/polymerase chain reaction and immunostaining, we investigated the gene expression and/or immunolocalization of endothelial NO synthase (eNOS), inducible NOS (iNOS), and nitrotyrosine (a biomarker for NO-derived peroxinitrite), and alkaline phosphatase (ALP) and osteocalcin (early and terminal differentiation markers of odontoblasts, respectively) in dental pulp tissue after rat tooth preparation. At the early stage (1-3 days) post-preparation, markedly increased expression of iNOS and nitrotyrosine was found in odontoblasts and pulp cells beneath the cavity, whereas eNOS expression was significantly decreased. ALP mRNA expression was significantly increased after 1 day but decreased after 3 days, whereas ALP activity was weak in the dentin-pulp interface under the cavity after 1 day but strong after 3 days. Osteocalcin mRNA expression was significantly increased at this stage. At 7 days post-preparation, tertiary dentin was formed under the cavity. All the molecules studied were expressed at control levels in odontoblasts/pulp cells beneath the cavity. These findings show that abundant NO is released from odontoblasts and pulp cells at an early stage after tooth preparation and indicate that, after tooth preparation, the up-regulation of iNOS and nitrotyrosine in odontoblasts is synchronized with increased cellular expression of ALP and osteocalcin. Therefore, the NO synthesized by iNOS after tooth preparation probably participates in regulating odontoblast differentiation during tertiary dentinogenesis.

DOI： 10.1007/s00441-005-0003-5

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1634/stemcells.2006-0576

Language：English  

DOI： 10.1002/9780470167526.ch8

Language：English  

DOI： 10.1002/9780470167526.ch8

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1371/journal.pone.0000079

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1371/journal.pone.0000079

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1111/j.1601-0825.2006.01300.x

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1111/j.1601-0825.2006.01300.x

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1007/s00441-006-0183-7

Language：English   Publishing type：Research paper (scientific journal)  

Capsaicin receptor expression in the rat temporomandibular joint.
Experimentally, temporomandibular joint (TMJ) nerve units respond to capsaicin, which is used clinically to treat TMJ pain. However, the existence of capsaicin receptors in the TMJ has not previously been clearly demonstrated. Immunohistochemical analysis has revealed the presence of transient receptor potential vanilloid subtype 1 (TRPV1) expression in the nerves and synovial lining cells of the TMJ. TRPV1-immunoreactive nerves are distributed in the synovial membrane of the joint capsule and provide branches to the joint compartment. The disc periphery is supplied by TRPV1 nerves that are mostly associated with small arterioles, and occasional nerves penetrate to the synovial lining layer. Double immunofluorescence has shown that many TRPV1-immunoreactive nerves are labeled with neuropeptide calcitonin gene-related peptide, whereas few are labeled with IB4-lectin. The results provide evidence for the presence of TRPV1 in both nerves and synovial lining cells, which might thus be involved in the mechanism of nociception and inflammation in the TMJ.

DOI： 10.1007/s00441-006-0183-7

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1679/aohc.68.259

Language：English   Publishing type：Research paper (scientific journal)  

Immunocytochemical localization of the neurokinin 1 receptor in rat dental pulp.
The dentin-pulp complex is a peripheral end-organ supplied by dense sensory nerve fibers. Substance P, a representative neuropeptide widely distributed in the dental pulp, has been reported to play roles in pain transmission and the amplification of inflammation. We analyzed here the expression of the neurokinin 1 (NK1) receptor, preferentially activated by substance P, using immunocytochemistry in rat dental pulp at both the light and electron microscopic levels. Conspicuous NK1 receptor immunoreactivity was found in the odontoblasts; immunolabelings were present at their plasma membrane and endosomal structures, especially in their cytoplasmic processes. Immunoreactions for NK1 receptor were also detectable in a part of the nerve terminals associated with the cytoplasmic processes of the odontoblasts. Furthermore, the endothelial cells of capillaries and post-capillary venules and the fibroblasts were labeled with the NK1 receptor in the subodontoblast layer. These findings suggest that pulpal cells and nerve fibers are targets for substance P that mediate multiple functions, including a vasoactive function and the regulation of vascular permeability as well as the modulation of pain transmission.

Language：English   Publishing type：Research paper (scientific journal)  

Laminin-5 (Ln-5) is an important molecule associated with epithelial cell adhesion and migration. In the gingiva around the tooth, Ln-5 localizes within basement membranes between the junctional epithelium (JE) and the tooth or connective tissue. Recently, we reported that in the oral mucosa around a dental implant, Ln-5 is expressed within the basement membranes at the implant-peri-implant epithelium (PIE) interface, and at the PIE-connective tissue interface. However, the ultrastructural localization of Ln-5 within or along the PIE has not yet been reported. Therefore, peri-implant oral mucosa was treated with anti-Ln-5 (γ2 chain) antibody and examined using immuno-electron microscopy. Ln-5 was localized in the cells of the innermost-third layer and basal layer of the PIE. A 100-nm-wide Ln-5-positive internal basal lamina (basement membrane) and hemidesmosomes as adhesion structures were formed at the apical portion of the implant-PIE interface. However, at the upper-middle portion of the interface, these adhesion structures were not observed. Furthermore, at the PIE-connective tissue interface, the Ln-5-positive external basal lamina (basement membrane) and hemidesmosomes were partially deficient. Judging from these findings, we concluded that Ln-5 contributes to the attachment of the PIE to the titanium surface, and that PIE attached to titanium at the apical portion of the dental implant-PIE interface.

DOI： 10.1016/j.biomaterials.2005.03.046

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.biomaterials.2004.05.033

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1267/ahc.38.121

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1007/s00441-004-1035-y

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1007/s00441-004-1035-y

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.biomaterials.2004.02.072

Language：Japanese  

Language：Japanese  

Language：Japanese  

Language：Japanese  

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1177/154405910408300807

Language：English   Publishing type：Research paper (scientific journal)  

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1093/jmicro/52.6.551

Language：Japanese  

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1177/154405910308200513

Language：English  

Language：English   Publishing type：Research paper (other academic)  

Language：English   Publishing type：Research paper (scientific journal)  

NF-κB plays a pivotal role in pathogenesis in general arthritis. However, the participation of NF-κB in inflammation of the temporomandibular joint (TMJ) is poorly understood. We examined NF-κB expression in rat TMJs with synovitis induced by condyle hypermobility. By immunohistochemistry, NF-κB immunoreactivity was found mainly in the cytoplasm, not the nucleus, of the synovial lining cells of induced-synovitis and control TMJs. Southwestern histochemistry, a new method for detecting transcription factors, showed greater NF-κB expression in the nucleus of the synovial lining cells in the hypertrophic synovium than in control synovium. Increased numbers of the synovial lining cells with immunoreactivity for inducible nitric oxide synthase (iNOS), which is transcriptionally regulated by NF-κB, were also seen in the inflamed synovium. These findings indicate that excess mechanical stress increases NF-κB activation in the TMJ and suggest that active NF-κB is involved in the progression of TMJ inflammation.

DOI： 10.1177/154405910308200307

Language：English  

NF-kB Activation and iNOS Expression in the Synovial Membrane of Rat Temporomandibular Joints after Induced Synovitis

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1093/jmicro/52.6.627

Language：Japanese  

Language：Japanese  

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1034/j.1600-0501.2002.130303.x

Language：English  

DOI： 10.1034/j.1600-0501.2002.130303.x

Language：English   Publishing type：Research paper (scientific journal)  

Nitric oxide (NO) is generated from L-arginine by NO synthase (NOS) and has multiple functions under both physiological and pathological conditions. One isoform of NOS, inducible NOS (iNOS) is expressed in the cells such as macrophages after induction with various cytokines or mechanical stress. In this study, we investigated the distribution of iNOS, interleukin-1β (IL-1β) and its receptor, the IL-1 receptor (IL-1R), in the synovial membrane of the temporomandibular joint (TMJ) of normal rats using immunolight and immunoelectron microscopy. By light microscopy, an immunopositive reaction for iNOS and IL-1β was found in the superficial cells of the synovial membrane of both the anterior and posterior portions of the articular disc. Immunoelectron microscopy revealed that iNOS-immunoreactive products were deposited in the cytoplasm and vesicles, and on the plasma membrane of type-A (macrophage-like) and B (fibroblast-like) cells of the superficial layer. IL-1R-positive products were found both on the plasma membrane and in the vesicles of type-A cells of the synovial lining, and were observed in macrophages in the sublining layer. These results reveal that iNOS and IL-1β localize to the synovial membrane of the rat TMJ under physiological conditions. Therefore, it is likely that autocrine/paracrine effects of IL-1β induce NO generation by iNOS via the IL-1R in type-A cells. It is considered that cytokine-induced NO may play an important role in the physiological maintenance, e.g. self-protection, by synovial lining cells of the synovial membrane in the TMJ.

DOI： 10.1267/ahc.35.11

Language：Japanese  

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1007/s007950170001

Language：English  

DOI： 10.1016/S8756-3282(01)00466-5

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1007/s004290100163

Language：English  

DOI： 10.1007/s004290100163

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1007/s004410000327

Language：Japanese  

Language：English  

DOI： 10.1007/s004410000327

Language：English   Publishing type：Research paper (scientific journal)  

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1267/ahc.34.31

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/S8756-3282(01)00466-5

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1267/ahc.34.31

Language：Japanese  

Language：English  

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1902/jop.2000.71.6.961

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1902/jop.2000.71.6.961

Language：Japanese  

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1007/s004410051349

Language：English  

DOI： 10.1007/s004410051349

Language：Japanese  

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/S8756-3282(98)00138-0

Language：English  

DOI： 10.1016/S8756-3282(98)00138-0

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1007/s004410051100

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1007/s004410051100

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1111/j.1600-0765.1997.tb00575.x

Language：Japanese  

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/S0003-9969(97)00003-4

Language：English  

DOI： 10.1016/S0003-9969(97)00003-4

Language：Japanese  

Language：Japanese  

Language：Japanese  

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1111/j.1600-0765.1996.tb00483.x

Language：English  

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/0003-9969(95)00060-3

Language：English  

Language：Japanese  

Language：Japanese  

Language：Japanese  

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1267/ahc.28.523

Responsible for pages：361-365   Language：Japanese   Book type：Scholarly book

Event date： 2017.10

Language：Japanese   Presentation type：Symposium, workshop panel (public)  

Venue：川崎   Country：Japan  

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Venue：福岡   Country：Japan  

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Venue：京都   Country：Japan  

Event date： 2014.2 - 2014.3

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Country：Japan  

Event date： 2013.9 - 2023.9

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Venue：岡山   Country：Japan  

Event date： 2013.9 - 2023.9

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Country：Japan  

Event date： 2013.9 - 2023.9

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Venue：岡山   Country：Japan  

Event date： 2013.9 - 2023.9

Language：Japanese   Presentation type：Symposium, workshop panel (public)  

Venue：岡山   Country：Japan  

Event date： 2013.9 - 2023.9

Language：Japanese   Presentation type：Symposium, workshop panel (public)  

Venue：岡山   Country：Japan  

Event date： 2013.9

Language：Japanese   Presentation type：Symposium, workshop panel (public)  

Venue：岡山   Country：Japan  

Event date： 2013.8 - 2023.8

Language：English   Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2013.7 - 2014.7

Language：Japanese   Presentation type：Symposium, workshop panel (public)  

Venue：京都   Country：Japan  

Event date： 2012.11

Presentation type：Oral presentation (general)  

Country：Taiwan, Province of China  

Event date： 2012.9

Presentation type：Symposium, workshop panel (public)  

Venue：郡山   Country：Japan  

Event date： 2012.9

Presentation type：Symposium, workshop panel (public)  

Venue：郡山   Country：Japan  

Event date： 2012.9

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Country：Japan  

Event date： 2012.8

Language：English   Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2012.5

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Country：Japan  

Event date： 2011.10

Presentation type：Symposium, workshop panel (public)  

Country：Japan  

Event date： 2011.9 - 2011.10

Presentation type：Symposium, workshop panel (public)  

Venue：長良川国際会議場   Country：Japan  

Event date： 2011.9 - 2011.10

Presentation type：Oral presentation (general)  

Venue：長良川国際会議場   Country：Japan  

Event date： 2011.9 - 2011.10

Presentation type：Symposium, workshop panel (public)  

Venue：岐阜   Country：Japan  

Event date： 2011.9 - 2011.10

Presentation type：Symposium, workshop panel (public)  

Venue：長良川国際会議場   Country：Japan  

Event date： 2011.9 - 2011.10

Presentation type：Oral presentation (general)  

Venue：長良川国際会議場   Country：Japan  

Event date： 2011.9 - 2011.10

Presentation type：Oral presentation (general)  

Venue：長良川国際会議場   Country：Japan  

Event date： 2011.9 - 2011.10

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.9

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.9

Presentation type：Symposium, workshop panel (public)  

Country：Japan  

Event date： 2011.8

Presentation type：Symposium, workshop panel (public)  

Country：China  

Event date： 2011.7

Presentation type：Symposium, workshop panel (public)  

Venue：大阪国際会議場   Country：Japan  

Event date： 2011.6

Presentation type：Oral presentation (general)  

Venue：大阪歯科大学   Country：Japan  

Event date： 2011.5

Presentation type：Symposium, workshop panel (public)  

Venue：福岡   Country：Japan  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：United States  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Romania  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.4

Presentation type：Symposium, workshop panel (public)  

Country：Japan  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：United States  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Romania  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.4

Presentation type：Symposium, workshop panel (public)  

Country：Japan  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.4

Presentation type：Symposium, workshop panel (public)  

Country：Japan  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2011.4

Country：Japan  

Event date： 2011.4

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Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：United States  

Event date： 2011.4

Presentation type：Oral presentation (general)  

Country：United States  

Event date： 2011.3

Presentation type：Symposium, workshop panel (public)  

Venue：神奈川歯科大学附属横浜研修センター   Country：Japan  

Event date： 2010.12

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Venue：広島   Country：Japan  

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Country：Japan  

Event date： 2010.7

Presentation type：Oral presentation (invited, special)  

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Presentation type：Symposium, workshop panel (public)  

Venue：盛岡   Country：Japan  

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Country：Japan  

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Event date： 2008.11

Presentation type：Symposium, workshop panel (public)  

Country：China  

Event date： 2008.11

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Country：China  

Event date： 2003.4

Presentation type：Symposium, workshop panel (public)  

Venue：福岡   Country：Japan  

Presentation type：Symposium, workshop panel (public)  

Venue：福岡   Country：Japan  

Presentation type：Symposium, workshop panel (public)  

Venue：福岡   Country：Japan  

Presentation type：Symposium, workshop panel (public)  

Venue：大阪   Country：Japan  

Presentation type：Oral presentation (general)  

Venue：北九州   Country：Japan  

Presentation type：Oral presentation (general)  

Venue：北九州   Country：Japan  

Presentation type：Symposium, workshop panel (public)  

Venue：大宮   Country：Japan  

Presentation type：Symposium, workshop panel (public)  

Venue：北九州   Country：Japan  

Presentation type：Oral presentation (general)  

Venue：福岡   Country：Japan  

Presentation type：Oral presentation (general)  

Venue：福岡   Country：Japan  

Presentation type：Oral presentation (general)  

Venue：鹿児島   Country：Japan  

Presentation type：Symposium, workshop panel (public)  

Country：Canada  

Presentation type：Oral presentation (general)  

Venue：福島   Country：Japan  

Presentation type：Oral presentation (general)  

Venue：大阪   Country：Japan  

Presentation type：Oral presentation (general)  

Venue：東京   Country：Japan  

Presentation type：Oral presentation (general)  

Venue：京都   Country：Japan  

Presentation type：Symposium, workshop panel (public)  

Country：United States  

Presentation type：Oral presentation (general)  

Venue：福岡   Country：Japan  

Presentation type：Symposium, workshop panel (public)  

Venue：東京   Country：Japan  

Presentation type：Oral presentation (general)  

Venue：横須賀   Country：Japan  

Presentation type：Symposium, workshop panel (public)  

Venue：横須賀   Country：Japan  

Presentation type：Symposium, workshop panel (public)  

Country：Japan  

Presentation type：Symposium, workshop panel (public)  

Venue：神戸   Country：Japan  

Presentation type：Symposium, workshop panel (public)  

Venue：さいたま市   Country：Japan  

Presentation type：Oral presentation (general)  

Venue：東京   Country：Japan  

Presentation type：Oral presentation (general)  

Venue：北九州   Country：Japan  

Presentation type：Oral presentation (general)  

Venue：東京   Country：Japan  

Presentation type：Oral presentation (general)  

Venue：東京   Country：Japan  

Presentation type：Symposium, workshop panel (public)  

Venue：東京   Country：Japan  

Presentation type：Symposium, workshop panel (public)  

Venue：名古屋  

Presentation type：Oral presentation (general)  

Venue：東京   Country：Japan  

Presentation type：Symposium, workshop panel (public)  

Country：Japan  

Presentation type：Oral presentation (general)  

Venue：福岡   Country：Japan  

Presentation type：Oral presentation (general)  

Venue：福岡   Country：Japan  

Presentation type：Symposium, workshop panel (public)  

Venue：盛岡   Country：Japan  

Presentation type：Oral presentation (general)  

Venue：盛岡   Country：Japan  

Presentation type：Oral presentation (general)  

Venue：盛岡   Country：Japan  

Presentation type：Symposium, workshop panel (public)  

Venue：岐阜   Country：Japan  

Presentation type：Oral presentation (general)  

Venue：東京   Country：Japan  

Presentation type：Oral presentation (general)  

Venue：東京   Country：Japan  

Presentation type：Oral presentation (general)  

Venue：大阪   Country：Japan  

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：横浜   Country：Japan  

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：福岡   Country：Japan  

Language：English   Presentation type：Symposium, workshop panel (public)  

Country：Japan  

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：福岡   Country：Japan  

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：横浜   Country：Japan  

Language：English  

Language：Japanese