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

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

CD14 and Toll-like receptor 4/MD2 (TLR4/MD2) mediate the action of LPS on neutrophils. The anti-CD14 antibody and the TLR4/MD2-antagonist, synthetic lipid IVa (LA-14-PP), are known to inhibit the response of neutrophils to LPS. We studied the role of CD14 in LPS-induced priming of neutrophils for enhanced release of the superoxide anion. The anti-CD14 antibody at much higher concentrations than required to saturate CD14 was required to inhibit priming by LPS. The inhibitory effect of the anti-CD14 antibody was overcome by LPS. After washing, anti-CD14-treated neutrophils showed upregulated CD14 upon incubation at 37°C and responded to LPS with a delayed time-course. Thus, CD14-blocked neutrophils gained responsiveness to LPS through newly upregulated CD14. These results suggested that the unbound/free anti-CD14 antibody was essential to inhibit LPS-induced priming by blocking CD14 that were newly expressed during incubation at 37°C. LA-14-PP inhibited the response of neutrophils to LPS in an anti-CD14 antibody sensitive manner. When neutrophils were treated with LA-14-PP followed by treatment with the anti-CD14 antibody, CD14 was upregulated upon warming, but priming was blocked, suggesting that TLR4/MD2 was not newly expressed by warming in association with CD14 molecules. Thus, in addition to blocking CD14, the anti-CD14 antibody was found to induce the expression of new CD14.

DOI： 10.1080/08820139.2016.1238925

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

DOI： 10.4172/jcs.1000117

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

Sprouty was identified as an inhibitor of the fibroblast growth factor (FGF) receptor, and Sprouty2 (Spry2) functions as a negative regulator of receptor tyrosine kinase signaling. In this study, we investigated how inhibition of Spry2 affects osteoblasts and gingival epithelial cells in periodontal tissue regeneration in vitro. Transduction of a dominant-negative mutant of Spry2 (Y55A-Spry2) enhanced basic fibroblast growth factor (bFGF)- and epidermal growth factor (EGF)-induced ERK activation in MC3T3-E1 osteoblastic cells. In contrast, it decreased their activation in GE1 cells. Consistent with these observations, Y55A-Spry2 increased osteoblast proliferation with bFGF and EGF stimulation, whereas the proliferation of Y55A-Spry2-introduced GE1 cells was decreased via the ubiquitination and degradation of EGF receptors (EGFRs). In addition, Y55A-Spry2 caused upregulation of Runx2 expression and downregulation of Twist, a negative regulator of Runx2, with treatment of bFGF and EGF, resulting in enhanced osteoblastogenesis accompanied by alkaline phosphatase activation and osteocalcin expression in MC3T3-E1 cells. These data suggest that suppression of Spry2 expression induces proliferation and differentiation of osteoblastic cells after the addition of a bFGF and EGF cocktail but inhibits proliferation in gingival epithelial cells. These in vitro experiments may provide a molecular basis for novel therapeutic approaches in periodontal tissue regeneration. Taken together, our study proposes that combined application of an inhibitor for tyrosine 55 of Spry2, bFGF, and EGF may effectively allow alveolar bone growth and block the ingrowth of gingival epithelial cells toward bony defects, biologically mimicking a barrier effect in guided tissue regeneration, with in vivo investigation in the future. J. Cell. Biochem. 116: 628-639, 2015.

DOI： 10.1002/jcb.25014

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DOI： 10.14800/rci.597

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Introduction: The purpose of this study was to examine the Streptococcus mutans biofilm cellular proteins recognized by immunoglobulin A (IgA) in saliva from various caries-defined populations. Methods: Biofilm and planktonic S. mutans UA159 cells were prepared. The proteins were extracted, separated by two-dimensional gel electrophoresis, transferred to blotting membranes, and probed for IgA using individual saliva samples from three groups of subjects; those who developed 0 caries (no active caries), 5-9 caries (medium), or more than 10 caries (severe) over a 12-month interval. Results: Several proteins were recognized by salivary IgA in all groups of saliva but spot distribution and intensity varied greatly between the groups, and some proteins were recognized more strongly in biofilm cells than in planktonic culture, and vice versa. Furthermore, 15 proteins were only recognized by saliva from the 'no active caries' group, and four proteins were recognized by saliva samples from subjects in all three groups. Specifically, antigen I/II was recognized less in biofilm cells by caries-free saliva compared with planktonic cells. However, salivary IgA antibody to antigen I/II was absent in blots using saliva from the 'medium caries' and 'severe caries' groups. Conclusion: The bacterial molecules recognized by caries-free saliva are significant factors for S. mutans caries formation, and their inhibition could be a therapeutic target. In addition, saliva of caries-free subjects includes significant IgA antibody against antigen I/II of S. mutans, indicating a protective mechanism. However, microorganisms may protect themselves from host immune attack by forming biofilms and decreasing expression of antigen I/II.

DOI： 10.1111/j.1399-302X.2009.00523.x

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

Although the migratory property of lymphocytes is critical for protective immunity, tissue infiltration of lymphocytes sometimes causes harmful immune responses. DOCK2 plays a critical role in lymphocyte migration by regulating actin cytoskeleton through Rac activation, yet the mechanism by which DOCK2 activates Rac remains unknown. We found that DOCK2 associates with engulfment and cell motility (ELMO1) through its Src-homology 3 (SH3) domain. When DOCK2 was expressed in T-hybridoma cells lacking endogenous expression of DOCK2, Rac activation and actin polymerization were induced. However, such responses were not elicited by the DOCK2 mutant lacking the region required for ELMO1 binding. On the other hand, we found that the expression of ELMO1 induces Rac activation in the plasmacytoma cells expressing DOCK2 but not ELMO1. These results indicate that the association of DOCK2 with ELMO1 is critical for DOCK2-mediated Rac activation, thereby suggesting that their association might be a therapeutic target for immunologic disorders caused by lymphocyte infiltration.

DOI： 10.1182/blood-2003-01-0173

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DOCK2 is a mammalian homolog of Caenorhabditis elegans CED-5 and Drosophila melanogaster Myoblast City which are known to regulate actin cytoskeleton. DOCK2 is critical for lymphocyte migration, yet the role of DOCK2 in TCR signaling remains unclear. We show here that DOCK2 is essential for TCR-mediated Rac activation and immunological synapse formation. In DOCK2-deficient T cells, antigen-induced translocation of TCR and lipid rafts, but not PKC-θ and LFA-1, to the APC interface was severely impaired, resulting in a significant reduction of antigen-specific T cell proliferation. In addition, we found that the efficacy of both positive and negative selection was reduced in DOCK2-deficient mice. These results suggest that DOCK2 regulates T cell responsiveness through remodeling of actin cytoskeleton via Rac activation.

DOI： 10.1016/S1074-7613(03)00169-9

Language：English   Publisher：Radiology Case Reports  

For maxillary gingival carcinomas, especially those in the molar region, surgical resection is often performed beyond the maxillary tuberosity. Bleeding from the posterior superior alveolar or maxillary artery into the pterygoid process is difficult to stop during partial maxillary resection. Advances in catheterization and materials have enabled the embolization of various vessels. In this report, we describe two cases of maxillary gingival cancer in which preoperative endovascular arterial embolization prevented bleeding due to unexpected vascular injury, allowing for a safe surgery with minimal blood loss. This technique effectively avoids emergency hemostasis for unexpected bleeding when resecting gingival cancers in the maxillary molar region.

DOI： 10.1016/j.radcr.2024.05.052

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Language：English   Publisher：American Journal of Case Reports  

Objective: Rare disease Background: Bisphosphonates and anti-receptor activator of nuclear factor kappa B antibodies are used to treat bone diseases associated with increased osteoclast activity, including myeloma. However, they can cause osteonecrosis of the jaw, known as medication-related osteonecrosis of the jaw. This report presents a case of a patient with a history of myeloma who required posterior maxilla resection for bisphosphonate-related osteonecrosis of the jaw, in which preoperative embolization prevented unexpected bleeding related to vascular injury and allowed for a safe procedure with minimal bleeding. Case Report: An 84-year-old man presented to our department with a 3-year history of purulent drainage and bone exposure in the right maxilla. Based on the clinical findings at the initial visit, the clinical diagnosis was bisphosphonate-related osteonecrosis of the jaw, and the patient underwent a partial right maxillary osteotomy. This surgery was associated with a risk of unexpected bleeding from a branch of the maxillary artery during the posterior maxilla resection. A catheter-based embolization of the maxillary artery was performed the day before performing a partial maxillectomy to avoid unexpected bleeding risk. Thus, no abnormal bleeding occurred during partial maxillectomy, and no postoperative complications occurred for 3 years. Conclusions: In the surgical treatment of medication-related osteonecrosis of the jaw, preoperative vascular embolization of the peripheral maxillary artery beyond the middle meningeal artery bifurcation is a valuable technique for safe maxillectomy involving the posterior maxilla.

DOI： 10.12659/AJCR.943807

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Biochemistry and Biophysics Reports  

Introduction & objectives: Stem cell therapy for regenerative medicine has been sincerely investigated, but not still popular although some clinical trials show hopeful results. This therapy is suggested to be a representative candidate such as bone defect due to the accident, iatrogenic resection oncological tumor, congenital disease, and severe periodontitis in oral region. Recently, the Bio-3D printer “Regenova®” has been introduced as an innovative three-dimensional culture system, equipped scaffold-free bio-assembling techniques without any biomaterials. Therefore, we expected a mount of bone defect could be repaired by the structure established from this Bio-3D printer using osteogenic potential stem cells. Material & methods: The gingival tissue (1x1 mm) was removed from the distal part of the lower wisdom tooth of the patients who agreed our study. Human Gingival Mesenchymal Stem Cells (hGMSCs) were isolated from this tissue and cultured, since we confirmed the characteristics such as facile isolation and accelerated proliferation, further, strong potential of osteogenic-differentiation. Spheroids were formed using hGMSC in 96-well plates designed for low cell adhesion. The size of the spheroids was measured, and fluorescent immunostaining was employed to verify the expression of stem cell and apoptosis marker, and extracellular matrix. Following four weeks of bone differentiation, μCT imaging was performed. Calcification was confirmed by alizarin red and von Kossa staining. Fluorescent immunostaining was utilized to assess the expression of markers indicative of advanced bone differentiation. Results: We have established and confirmed the spheroids (∼600 μm in diameter) constructed from human GMSCs (hGMSCs) still maintain stem cell potentials and osteogenic differentiation abilities from the results that CD73 and not CD34 were expressed as stem cell positive and negative marker, respectively. These spheroids were pilled up like cylindal shape to the “Kenzan” platform of Bio-3D printer and cultured for 7days. The cylindal structure originated from compound spheroids were tried to differentiate into bone four weeks with osteogenic induction medium. The calcification of bio-3D printed bone-like structures was confirmed by alizarin red and Von Kossa staining. In addition, μCT analysis revealed that the HU (Hounsfield Unit) of the calcified structures was almost identical to that of trabecular bone. Immunofluorescent staining detected osteocalcin expression, a late-stage bone differentiation marker. Conclusion: For the first time, we have achieved the construction of a scaffold-free, bone-like luminal structure through the assembly of spheroids comprised of this hGMSCs. This success is sure to be close to the induction of clinical application against regenerative medicine especially for bone defect disease.

DOI： 10.1016/j.bbrep.2024.101656

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Language：English   Publisher：Hindawi  

Aim. The present study evaluated the therapeutic effects of luteolin in alleviating pulpitis of dental pulp- (DP-) derived microvesicles (MVs) via the inhibition of protein kinase R- (PKR-) mediated inflammation. Methodology. Proteomic analysis of immortalized human dental pulp (DP-1) cell-derived MVs was performed to identify PKR-associated molecules. The effect of luteolin on PKR phosphorylation in DP-1 cells and the expression of tumor necrosis factor-α (TNF-α) in THP-1 macrophage-like cells were validated. The effect of luteolin on cell proliferation was compared with that of chemical PKR inhibitors (C16 and 2-AP) and the unique commercially available sedative guaiacol-parachlorophenol. In the dog experimental pulpitis model, the pulps were treated with (1) saline, (2) guaiacol-parachlorophenol, and (3) luteolin. Sixteen teeth from four dogs were extracted, and the pulp tissues were analyzed using hematoxylin and eosin staining. Immunohistochemical staining was performed to analyze the expression of phosphorylated PKR (pPKR), myeloperoxidase (MPO), and CD68. Experimental endodontic-periodontal complex lesions were established in mouse molar through a silk ligature and simultaneous MV injection. MVs were prepared from DP-1 cells with or without pretreatment with 2-AP or luteolin. A three-dimensional microcomputed tomography analysis was performed on day 7 (n=6). Periodontal bone resorption volumes were calculated for each group (nonligated–ligated), and the ratio of bone volume to tissue volume was measured. Results. Proteomic analysis identified an endogenous PKR activator, and a protein activator of interferon-induced PKR, also known as PACT, was included in MVs. Luteolin inhibited the expressions of pPKR in DP-1 cells and TNF-α in THP-1 cells with the lowest suppression of cell proliferation. In the dog model of experimental pulpitis, luteolin treatment suppressed the expression of pPKR-, MPO-, and CD68-positive cells in pulp tissues, whereas guaiacol-parachlorophenol treatment caused coagulative necrosis and disruption. In a mouse model of endodontic-periodontal complex lesions, luteolin treatment significantly decreased MV-induced alveolar bone resorption. Conclusion. Luteolin is an effective and safe compound that inhibits PKR activation in DP-derived MVs, enabling pulp preservation.

CiNii Research

Language：English   Publishing type：Research paper (scientific journal)   Publisher：BioMed Research International  

Aim. The present study evaluated the therapeutic effects of luteolin in alleviating pulpitis of dental pulp- (DP-) derived microvesicles (MVs) via the inhibition of protein kinase R- (PKR-) mediated inflammation. Methodology. Proteomic analysis of immortalized human dental pulp (DP-1) cell-derived MVs was performed to identify PKR-associated molecules. The effect of luteolin on PKR phosphorylation in DP-1 cells and the expression of tumor necrosis factor-α (TNF-α) in THP-1 macrophage-like cells were validated. The effect of luteolin on cell proliferation was compared with that of chemical PKR inhibitors (C16 and 2-AP) and the unique commercially available sedative guaiacol-parachlorophenol. In the dog experimental pulpitis model, the pulps were treated with (1) saline, (2) guaiacol-parachlorophenol, and (3) luteolin. Sixteen teeth from four dogs were extracted, and the pulp tissues were analyzed using hematoxylin and eosin staining. Immunohistochemical staining was performed to analyze the expression of phosphorylated PKR (pPKR), myeloperoxidase (MPO), and CD68. Experimental endodontic-periodontal complex lesions were established in mouse molar through a silk ligature and simultaneous MV injection. MVs were prepared from DP-1 cells with or without pretreatment with 2-AP or luteolin. A three-dimensional microcomputed tomography analysis was performed on day 7 (n=6). Periodontal bone resorption volumes were calculated for each group (nonligated-ligated), and the ratio of bone volume to tissue volume was measured. Results. Proteomic analysis identified an endogenous PKR activator, and a protein activator of interferon-induced PKR, also known as PACT, was included in MVs. Luteolin inhibited the expressions of pPKR in DP-1 cells and TNF-α in THP-1 cells with the lowest suppression of cell proliferation. In the dog model of experimental pulpitis, luteolin treatment suppressed the expression of pPKR-, MPO-, and CD68-positive cells in pulp tissues, whereas guaiacol-parachlorophenol treatment caused coagulative necrosis and disruption. In a mouse model of endodontic-periodontal complex lesions, luteolin treatment significantly decreased MV-induced alveolar bone resorption. Conclusion. Luteolin is an effective and safe compound that inhibits PKR activation in DP-derived MVs, enabling pulp preservation.

DOI： 10.1155/2024/8864513

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Publisher：Current Oral Health Reports  

Purpose of Review: Luteolin, a natural polyphenolic flavone, is a bioactive compound with high thermal stability. Owing to its prominent antioxidant activity, luteolin has been reported to exert therapeutic effects on inflammation-associated diseases. This review discusses the therapeutic potential of luteolin for treating dental diseases. Recent Findings: Luteolin has multifaceted pharmacological activities, including anti-inflammatory, neuroprotective, anticancer, and cardioprotective effects. Furthermore, the antibacterial effects of luteolin are accompanied by an anti-biofilm effect. More recently, luteolin has been identified as an inhibitor of protein kinase R (PKR), which plays an essential role in inflammasome activation. In this regard, we demonstrated the potential of luteolin as a pulp sedation compound for pulpitis that acts by suppressing PKR-mediated inflammation in dental pulp cells. Summary: Although conventional dental treatments for dental caries or periodontitis largely depend on cause-related therapy, disruption of biofilms and regulation of inflammation are prerequisites for a favorable prognosis. Together with its superior anti-inflammatory and antibacterial effects, the biocompatible features of luteolin make it a promising candidate for treating dental diseases with fewer side effects.

DOI： 10.1007/s40496-024-00389-w

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Publisher：Frontiers Media SA  

Drug-induced gingival overgrowth (DIGO), induced by certain immunosuppressive drugs, antihypertensive agents, and antiepileptic drugs, may contribute to the formation of deeper periodontal pockets and intractableness in periodontitis. To date, multiple factors such as enhanced matrix production, inflammation, and reduced matrix degradation might be involved in the pathogenesis of DIGO. We have previously reported that SPOCK-1, a heparan sulfate proteoglycan, could affect gingival thickening by promoting epithelial-to-mesenchymal transition (EMT) in gingival keratinocytes. However, few studies have investigated whether a combination of these factors enhances the DIGO phenotype in animal models. Therefore, we investigated whether SPOCK-1, periodontal inflammation, and cyclosporin-A (CsA) could cooperatively promote gingival overgrowth. We first confirmed that Spock-1 overexpressing (Spock1-Tg) mice showed significantly thicker gingiva and greater alveolar bone loss than WT mice in response to ligature-induced experimental periodontitis. DIGO was induced by the combination of CsA administration and experimental periodontitis was significantly enhanced in Spock1-Tg mice compared to that in WT mice. Ligature-induced alveolar bone loss in CsA-treated Spock1-Tg mice was also significantly greater than that in CsA-treated WT mice, while being accompanied by an increase in Rankl and Col1a1 levels and a reduction in matrix metalloprotease expression. Lastly, SPOCK-1 promoted RANKL-induced osteoclast differentiation in both human peripheral blood mononuclear cells and murine macrophages, while peritoneal macrophages from Spock1-Tg mice showed less TNFα and IL-1β secretion than WT mice in response to Escherichia coli lipopolysaccharide. These results suggest that EMT, periodontal inflammation, and subsequent enhanced collagen production and reduced proteinase production contribute to CsA-induced DIGO pathogenesis.

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

DOI： 10.7759/cureus.50228

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Frontiers in Physiology  

Drug-induced gingival overgrowth (DIGO), induced by certain immunosuppressive drugs, antihypertensive agents, and antiepileptic drugs, may contribute to the formation of deeper periodontal pockets and intractableness in periodontitis. To date, multiple factors such as enhanced matrix production, inflammation, and reduced matrix degradation might be involved in the pathogenesis of DIGO. We have previously reported that SPOCK-1, a heparan sulfate proteoglycan, could affect gingival thickening by promoting epithelial-to-mesenchymal transition (EMT) in gingival keratinocytes. However, few studies have investigated whether a combination of these factors enhances the DIGO phenotype in animal models. Therefore, we investigated whether SPOCK-1, periodontal inflammation, and cyclosporin-A (CsA) could cooperatively promote gingival overgrowth. We first confirmed that Spock-1 overexpressing (Spock1-Tg) mice showed significantly thicker gingiva and greater alveolar bone loss than WT mice in response to ligature-induced experimental periodontitis. DIGO was induced by the combination of CsA administration and experimental periodontitis was significantly enhanced in Spock1-Tg mice compared to that in WT mice. Ligature-induced alveolar bone loss in CsA-treated Spock1-Tg mice was also significantly greater than that in CsA-treated WT mice, while being accompanied by an increase in Rankl and Col1a1 levels and a reduction in matrix metalloprotease expression. Lastly, SPOCK-1 promoted RANKL-induced osteoclast differentiation in both human peripheral blood mononuclear cells and murine macrophages, while peritoneal macrophages from Spock1-Tg mice showed less TNFα and IL-1β secretion than WT mice in response to Escherichia coli lipopolysaccharide. These results suggest that EMT, periodontal inflammation, and subsequent enhanced collagen production and reduced proteinase production contribute to CsA-induced DIGO pathogenesis.

DOI： 10.3389/fphys.2023.1298813

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Journal of Dental Research  

Epidemiological studies suggest that the severity of periodontitis is higher in people with diabetes than in healthy individuals. Insulin resistance might play a crucial role in the pathogenesis of multiple diabetic complications and is reportedly induced in the gingiva of rodents with type 2 diabetes; however, the molecular mechanisms underlying the pathogenesis of diabetes-related periodontitis remain unclear. Therefore, we aimed to investigate whether endothelial insulin resistance in the gingiva may contribute to the pathogenesis of periodontitis as well as elucidate its underlying molecular mechanisms. We demonstrated that insulin treatment downregulated lipopolysaccharide (LPS)–induced or tumor necrosis factor α (TNFα)–induced VCAM1 expression in endothelial cells (ECs) via the PI3K/Akt activating pathway, resulting in reduced cellular adhesion between ECs and leukocytes. Hyperglycemia-induced selective insulin resistance in ECs diminished the effect of insulin on LPS- or TNFα-stimulated VCAM1 expression. Vascular endothelial cell–specific insulin receptor knockout (VEIRKO) mice exhibited selective inhibition of the PI3K/Akt pathway in the gingiva and advanced experimental periodontitis-induced alveolar bone loss via upregulation of Vcam1, Tnfα, Mcp-1, Rankl, and neutrophil migration into the gingiva compared with that in the wild-type (WT) mice despite being free from diabetes. We also observed that insulin-mediated activation of FoxO1, a downstream target of Akt, was suppressed in the gingiva of VEIRKO and high-fat diet (HFD)–fed mice, hyperglycemia-treated ECs, and primary ECs from VEIRKO. Further analysis using ECs transfected with intact and mutated FoxO1, with mutations at 3 insulin-mediated phosphorylation sites (T24A, S256D, S316A), suggested that insulin-mediated regulation of VCAM1 expression and cellular adhesion of ECs with leukocytes was attenuated by mutated FoxO1 overexpression. These results suggest that insulin resistance in ECs may contribute to the progression of periodontitis via dysregulated VCAM1 expression and cellular adhesion with leukocytes, resulting from reduced activation of the PI3K/Akt/FoxO1 axis.

DOI： 10.1177/00220345231181539

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Archives of Biochemistry and Biophysics  

A well-tuned inflammatory response is crucial for an effective immune process. Nuclear factor-kappa B (NF-κB) is a key mediator of inflammatory and innate immunity responses, and its dysregulation is closely associated with immune-related diseases. MicroRNAs (miRNAs) are important inflammation modulators. However, miRNA-regulated mechanisms that implicate NF-κB activity are not fully understood. This study aimed to identify a potential miRNA that could modulate the dysregulated NF-κB signaling during inflammation. We identified miR-582-5p that was significantly downregulated in inflamed murine adipose tissues and RAW264.7 cells. S-phase kinase-associated protein 1 (SKP1), a core component of an E3 ubiquitin ligase that regulates the NF-κB pathway, was proposed as a biological target of miR-582-5p by using TargetScan. The binding of miR-582-5p to a 3′-untranslated region site on Skp1 was confirmed using a dual-luciferase reporter assay; in addition, transfection with a miR-582-5p mimic suppressed SKP1 expression in RAW264.7 cells. Importantly, exogenous miR-582-5p attenuated the production of pro-inflammatory cytokines such as tumor necrosis factor-alpha, interleukin-1 beta, and interleukin-6 through suppressing the degradation of the NF-κB inhibitor alpha, followed by the nuclear translocation of NF-κB. Therefore, exogenously applied miR-582-5p can attenuate the NF-κB signaling pathway via targeting Skp1; this provides a prospective therapeutic strategy for treating inflammatory and immune diseases.

DOI： 10.1016/j.abb.2022.109501

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Language：English   Publisher：Frontiers Media SA  

The expression profiles of exosomal microRNAs (miRNAs) are regulated by the microenvironment, and appropriate priming with mesenchymal stem cells (MSCs) is one of the strategies to enhance the paracrine potency of MSCs. Our previous work demonstrated that exosomes from tumor necrosis factor (TNF)-α-primed human gingiva-derived MSCs (GMSCs) could be a therapeutic tool against periodontitis, and that TNFα-inducible exosomal miR-1260b is essential for the inhibition of alveolar bone loss. However, the precise molecular mechanism underlying miR-1260b-mediated inhibition of osteoclastogenesis is not yet fully understood. Here, we found that the activating transcription factor (ATF)-6β, a novel miR-1260b-targeting gene, is critical for the regulation of osteoclastogenesis under endoplasmic reticulum (ER) stress. An experimental periodontal mouse model demonstrated that induction of ER stress was accompanied by enhanced ATF6β expression, and local administration of miR-1260b and ATF6β siRNA using polyethylenimine nanoparticles (PEI-NPs) significantly suppressed the periodontal bone resorption. In periodontal ligament (PDL) cells, the ER stress inducer, tunicamycin, enhanced the expression of the receptor activator of NF-κB ligand (RANKL), while miR-1260b-mediated downregulation of ATF6β caused RANKL inhibition. Furthermore, the secretome from miR-1260b/ATF6β-axis-activated PDL cells inhibited osteoclastogenesis in human CD14^+ peripheral blood-derived monocytes. These results indicate that the miR-1260b/ATF6β axis mediates the regulation of ER stress, which may be used as a novel therapeutic strategy to treat periodontal disease.

DOI： 10.3389/fcell.2022.1061216

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DOI： 10.3389/icell.2022.1061216

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DOI： doi: 10.3389/fcell.2022.1061216. eCollection 2022.

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DOI： doi: 10.1007/s00592-022-01930-y.

Language：English   Publisher：Acta Diabetologica  

Aims: Pancreatic β-cell apoptosis may be involved in the onset and progression of type 2 diabetes mellitus, although its mechanism remains unclear. We previously demonstrated that macrophage-derived interferon (IFN) β induced X-linked inhibitor of apoptosis–associated factor 1 (XAF1) expression in β-cells and accelerated β-cell apoptosis in vitro. Here, we explored the effects of XAF1 on β-cell function and progression of diabetes in vivo. Methods: Pancreatic β-cell-selective XAF1 overexpressing (Xaf1 Tg) mice were generated. Xaf1 Tg mice and their wild-type (WT) littermates were fed either a normal diet or a 40% or 60% high-fat diet (HFD). The effects of β-cell XAF1 on β-cell apoptosis and exacerbation of diabetes were investigated. Results: Palmitic acid induced IFNβ expression in macrophages, and HFD intake promoted macrophage infiltration in pancreatic islets, both of which cooperatively upregulated XAF1 expression in mouse islets. Furthermore, HFD-fed Xaf1 Tg mice demonstrated increased β-cell apoptosis, lowered insulin expression, and impaired glucose tolerance compared with WT mice fed the same diet. These effects were more pronounced in the 60%HFD group than in the 40%HFD group. Conclusions: Pancreatic β-cell XAF1 expression was enhanced via HFD-induced, macrophage-derived IFNβ, which promoted β-cell apoptosis and led to a reduction in insulin secretion and progression of diabetes. To our knowledge, this is the first report to demonstrate an association between pancreatic β-cell XAF1 overexpression and exacerbation of diabetes, thus providing insight into the mechanism of β-cell mass reduction in diabetes.

DOI： 10.1007/s00592-022-01930-y

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Publisher：Oral and Maxillofacial Surgery Cases  

Basal cell adenoma (BCA) is a relatively rare benign tumor of the salivary glands. However, BCA rarely occurs in the minor salivary glands. Herein, we report a rare case of BCA that originated in the left buccal mucosa of a 47-year-old female. Magnetic resonance imaging revealed a mass with a diameter of 1.5 cm and a well-defined periphery in a high-signal area on a T2-weighted image. The mass was located between the buccal and masseter muscles on the left side of the face. A biopsy was performed, and the results indicated a pathological diagnosis of BCA. Thereafter, excision of the mass was performed under local anesthesia. Histopathological examination showed that the tumor was covered with a fibrous capsule and filled with basal cell-like cells that proliferated in a solid, tubular, and trabecular pattern, with no cellular atypia or infiltrative growth. Immunohistochemistry analysis revealed that the tumor cells inside the luminal-tubular structure showed positive immunoreactivity for CK1/3, whereas those outside expressed αSMA and vimentin. However, most of the tumor cells were p63-positive. The Ki-67 labeling index of the tumor was 3%. In addition to this report, we present a review of cases of salivary gland tumors reported from 1991 to date. The review of the 20 cases (including the present case) indicated that the most common sites of occurrence were the upper lip and palate (7 [35%] cases), buccal mucosa (4 [20%] cases), and alveolar region (2 [10%] cases). The most common histopathological type was the solid type (12 cases).

DOI： 10.1016/j.omsc.2022.100276

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Language：English   Publisher：Springer  

Immunoregulatory properties of mesenchymal stem cell (MSC)-derived extracellular vesicles (EVs) are promising. Gingival tissue-derived MSCs (GMSCs) have unique immunoregulatory capacity and secrete large amounts of EVs. Recent findings suggest that priming MSCs with inflammatory stimuli is an effective strategy for cell-free therapy. However, the precise mechanism by which the contents of EVs are customized has not been fully elucidated. Here, we show that EVs derived from GMSCs primed with a combination of two pro-inflammatory cytokines, tumor necrosis factor-α (TNF-α) and interferon-α (IFN-α), synergistically promote anti-inflammatory M2 macrophage polarization by increasing the expression of cluster of differentiation 73 (CD73) and CD5 molecule-like (CD5L). Expression of CD73 by TNF-α/IFN-α stimulation was transcriptionally upregulated by the activation of mammalian target of rapamycin signaling and nuclear translocation of hypoxia-inducible factor 1α in GMSCs. TNF-α/IFN-α treatment also significantly increased the expression of CD5L mRNA via the transcription factor DNA-binding protein inhibitor ID3 and liver X receptor. Interestingly, exosomal CD5L is a prerequisite for the synergistic effect of EVs-mediated M2 macrophage polarization. These results indicate that combined pre-licensing with TNF-α and IFN-α in GMSCs is ideal for enhancing the anti-inflammatory function of EVs, which contributes to the establishment of a therapeutic tool.

DOI： 10.1038/s41598-022-17692-0

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DOI： doi: 10.1038/s41598-022-17692-0.

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DOI： doi: 10.1136/bmjdrc-2020-001871.

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DOI： doi: 10.1016/j.actbio.2020.12.046.

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DOI： doi: 10.1016/j.bbrc.2020.09.129.

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DOI： doi: 10.1038/s41598-020-66660-z.

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DOI： 10.1002/cbin.10876.

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

Objectives Amelogenin, the major component of the enamel matrix derivative (EMD), has been suggested as a bioactive candidate for periodontal regeneration. Apart from producing a regenerative effect on periodontal tissues, amelogenin has also been reported to have an anti-inflammatory effect. However, the precise molecular mechanisms underlying these effects remain unclear. In the present study, we examined the immunomodulatory effects of amelogenin on macrophages. Design Human phorbol 12-myristate 13-acetate (PMA)-differentiated U937 macrophages and CD14+ peripheral blood-derived monocytes (PBMC)-derived macrophages were stimulated with recombinant amelogenin (rM180). After performing a detailed microarray analysis, the effects of rM180 on macrophage phenotype and signal transduction pathways were evaluated by real-time polymerase chain reaction, enzyme-linked immunosorbent assay, confocal microscopy and flow cytometry. Results The microarray analysis demonstrated that rM180 increased the expression of anti-inflammatory genes in lipopolysaccharide (LPS)-challenged macrophages after 24 h, while it temporarily up-regulated inflammatory responses at 4 h. rM180 significantly enhanced the expression of M2 macrophage markers (CD163 and CD206). rM180-induced M2 macrophage polarisation was associated with morphological changes as well as vascular endothelial growth factor (VEGF) production. rM180 enhanced prostaglandin E2 (PGE2) expression, and the activation of the cAMP/cAMP-responsive element binding (CREB) signaling pathway was involved in amelogenin-induced M2 macrophage polarisation. Blocking of PGE2 signaling by indomethacin specifically abrogated rM180 with or without LPS-induced M2 shift in PBMC-derived macrophages. Conclusion Amelogenin could reprogram macrophages into the anti-inflammatory M2 phenotype. It could therefore contribute to the early resolution of inflammation in periodontal lesions and provide a suitable environment for remodeling-periodontal tissues.

DOI： 10.1016/j.archoralbio.2017.08.005

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

Introduction and Aims Several studies have reported that angiopoietin-like protein 2 (Angptl2) is expressed abundantly in adipocytes and is associated with adipose tissue inflammation. In the present study, we found that osteoblasts and mesenchymal stem cells also expressed Angptl2 at high levels. The aim of this study was to understand the role of Angptl2 in osteoblastic cell differentiation. Methods Angptl2 expression was examined during osteoblast and adipocyte differentiation. The role of Angptl2 on cell differentiation and associated signaling was analyzed by gene knockdown using Angptl2 small interfering ribonucleic acid (siRNA). Results Angptl2 was highly expressed in MC3T3-E1 cells, ST2 cells and primary osteoblasts, but not in RAW264 cells. Inhibition of Angptl2 expression using siRNA markedly inhibited alkaline phosphatase (ALP) expression and osteoblastic differentiation in MC3T3-E1, ST2 cells and primary osteoblasts. Angptl2 siRNA also inhibited adipocyte differentiation in ST2 cells. Treatment of MC3T3-E1 cells with Angptl2 siRNA led to the down-regulation of the activities of several cell signaling pathways, including extracellular signal-regulated kinase (ERK), Jun amino-terminal kinase (JNK), Akt, and nuclear factor kappa B (NF-κB) signals. It also down-regulated the expression of Osterix, but not that of runt-related transcription factor 2 (Runx2), suggesting that Angptl2 is a positive activator of Osterix and its down-stream signals. Treatment of MC3T3-E1 cells with anti-Angptl2 antibodies suppressed ALP gene expression. In addition, treatment of Angptl2 siRNA-treated cells with culture supernatants of normal MC3T3-E1 cells restored ALP gene expression, indicating that Angptl2 acts in an autocrine manner. Conclusions The results suggest that Angptl2 is an autocrine positive regulator of cell differentiation. Thus, it is suggested that Angptl2 regulates not only adipose tissue metabolism but also bone metabolism.

DOI： 10.1016/j.metabol.2017.01.006

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

Background and aims Epicatechin (EC) intake has been suggested to be beneficial for the prevention of cardiovascular disorders, and it is well known that adipose tissue inflammation is one of the major risk factors for coronary heart diseases. The purpose of the present study was to determine the in vitro and in vivo effects of EC on adipose tissue inflammation and obesity. Methods and results DNA microarray analysis was performed to evaluate the effects of EC on gene expression in adipocytes co-cultured with bacterial endotoxin-stimulated macrophages. To determine the in vivo effects of the catechin, C57BL/6 mice were fed either a high-fat diet (HFD) or HFD combined with EC, and metabolic changes were observed EC suppressed the expression of many inflammatory genes in the adipocytes co-cultured with endotoxin-stimulated macrophages. Specifically, EC markedly suppressed chemokine (C–C motif) ligand 19 (CCL19) expression. The target cell of EC appeared to macrophages. The in vivo study indicated that mice fed the EC-supplemented HFD were protected from diet-induced obesity and insulin resistance. Accordingly, the expression levels of genes associated with inflammation in adipose tissue and in the liver were downregulated in this group of mice. Conclusions EC exerts beneficial effects for the prevention of adipose tissue inflammation and insulin resistance. Since we previously reported that mice deficient in the CCL19 receptor were protected from diet-induced obesity and insulin resistance, it can be concluded that the beneficial effects of EC could be mediated, at least in part, by marked suppression of CCL19 expression.

DOI： 10.1016/j.numecd.2016.11.008

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

DOI： doi:10.1007/s.40496-017-0136-0

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

Mechanical reduction of infectious bacteria by using physical instruments is considered the principal therapeutic strategy for periodontal disease; addition of antibiotics is adjunctive. However, local antibiotic treatment, combined with conventional mechanical debridement, has recently been shown to be more effective in periodontitis subjects with type 2 diabetes. This suggests that some bacteria may invade the inflamed inner gingival epithelium, and mechanical debridement alone will be unable to reduce these bacteria completely. Therefore, we tried to establish infected organ culture models that mimic the inner gingival epithelium and aimed to see the effects of antibiotics in these established models. Mouse dorsal skin epithelia were isolated, and periodontal bacteria were injected into the epithelia. Infected epithelia were incubated with test antibiotics, and colony-forming ability was evaluated. Results indicated that effective antibiotics differed according to injected bacteria and the bacterial combinations tested. Overall, in organ culture model, the combination of amoxicillin or cefdinir and metronidazole compensate for the effects of less effective bacterial combinations on each other. This in vitro study would suggest effective periodontal treatment regimens, especially for severe periodontitis.

DOI： 10.1002/cre2.48

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

Fimbriae are virulence factors of Porphyromonas gingivalis (P. gingivalis). In this study, the action of fimbriae on neutrophil respiratory burst and cytokine production by mononuclear cells (MNC) were investigated. Native or denatured form of purified P. gingivalis fimbriae contained endotoxin at an equivalence of 1– 3 μg lipopolysaccharides (LPS)/mg protein. The endotoxin could be reduced to the equivalent of 1 ng-LPS/mg protein by phase separation using Triton X-114. Unfractionated fimbriae caused serum-dependent priming of neutrophils for enhanced respiratory burst, but both native and denatured forms of Triton X-114-fractionated fimbriae were not active at 100 μg/mL. Unfractionated fimbriae induced serum-dependent production of IL-1β by MNC. Triton X-114-fractionated fimbriae (10 μg/mL)-induced production of IL-1β, IL-8 or TNF-α was much lower than that induced by unfractionated fimbriae or 10 ng/mL P. gingivalis-LPS preparation. Triton X-114-fractionated fimbriae immobilized on polystyrene tubes induced adhesion-stimulated superoxide release by LPS-primed neutrophils in a β₂ integrin-dependent manner. P. gingivalis cells caused priming of neutrophils; however, Toll-like receptor (TLR) 4 antagonists did not affect this response. Thus, P. gingivalis fimbriae were ineffective in inducing innate immune response in leukocytes; however, they induced β₂ integrin-mediated response by neutrophils. Immune-stimulatory components of P. gingivalis might be recognized by receptors other than TLR4.

DOI： 10.1016/j.jim.2016.11.012

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

Fibroblast growth factor (FGF) and bone morphogenetic protein (BMP) play essential roles in bone formation and osteoblast activity through the extracellular signal-regulated kinase 1/2 (ERK1/2) and Smad pathways. Sprouty family members are intracellular inhibitors of the FGF signaling pathway, and four orthologs of Sprouty have been identified in mammals. In vivo analyses have revealed that Sprouty2 is associated with bone formation. However, the mechanism by which the Sprouty family controls bone formation has not been clarified. In this study, we investigated the involvement of Sprouty2 in osteoblast proliferation and differentiation. We examined Sprouty2 expression in MC3T3-E1 cells, and found that high levels of Sprouty2 expression were induced by basic FGF stimulation. Overexpression of Sprouty2 in MC3T3-E1 cells resulted in suppressed proliferation compared with control cells. Sprouty2 negatively regulated the phosphorylation of ERK1/2 after basic FGF stimulation, and of Smad1/5/8 after BMP stimulation. Furthermore, Sprouty2 suppressed the expression of osterix, alkaline phosphatase, and osteocalcin mRNA, which are markers of osteoblast differentiation. Additionally, Sprouty2 inhibited osteoblast matrix mineralization. These results suggest that Sprouty2 is involved in the control of osteoblast proliferation and differentiation by downregulating the FGF-ERK1/2 and BMP-Smad pathways, and suppresses the induction of markers of osteoblast differentiation.

DOI： 10.1002/cbin.10876

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

Interleukin-17A (IL-17A) is known to induce inflammatory responses and to be involved in the pathogenesis of not only autoimmune diseases, but also several metabolic and infectious diseases. In this study, IL-17A is shown to induce IL-6 expression in 3T3-L1 mature adipocytes. Interestingly, we found that IL-17A synergistically amplified TNFα-induced secretion of IL-6 and upregulation of IL-17RA expression in 3T3-L1 adipocytes. Its synergistic effects on IL-6 production were inhibited by pre-treatment with inhibitors of IκBα and JNK. Furthermore, IL-17A cooperatively enhanced LPS-mediated IL-6 production in 3T3-L1 adipocytes co-cultured with RAW264.7 macrophages. In addition, IL-17A also enhanced CCL20 production in 3T3-L1 adipocytes stimulated with TNFα or co-cultured with LPS-stimulated RAW macrophages. In high-fat diet-fed mouse epididymal adipose tissues, IL-17RA and RORγt mRNA levels were significantly increased and the serum level of CCL20 was also upregulated. Taken together, these data show that, in adipose tissues, IL-17A contributes to exacerbating insulin resistance-enhancing IL-6 production and promotes the infiltration of Th17 cells in cooperation with TNFα; these findings represent a novel hypothesis for the association between IL-17A-producing cells and type 2 diabetes.

DOI： 10.1016/j.bbrc.2016.06.049

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

The effect of macrolides on the superoxide (O₂ ⁻) production by neutrophils was studied. Resting neutrophils become primed by lipopolysaccharide (LPS) or N-formyl-methionyl-leucyl-phenylalanine (fMLP), and primed neutrophils generate O₂ ⁻ in response to fMLP or adhesion, respectively. Both LPS-primed fMLP-stimulated O₂ ⁻ generation by macrolide-treated neutrophils and adhesion-stimulated O₂ ⁻ generation by macrolide-treated fMLP-primed neutrophils were inhibited. Macrolide inhibition of O₂ ⁻ generation was dependent on serum or pH. Serum could be substituted by NaHCO₃. The intensity of inhibition was azithromycin = roxithromycin > clarithromycin > erythromycin, in that order. Non-antimicrobial derivatives of erythromycin, that is, EM703 and EM900, inhibited O₂ ⁻ generation at pH 7.4. NH₄Cl abolished the activity of azithromycin (AZ) only when added to neutrophils with AZ but not after incubation with AZ, suggesting that NH₄Cl prevented the influx of AZ. AZ did not affect the expression of alkaline phosphatase, CD11b, and cytochrome b₅₅₈ in both resting and LPS-primed neutrophils. These results suggested that macrolides did not affect granule mobilization but inhibited O₂ ⁻ generation selectively.

DOI： 10.1007/s10753-016-0333-3

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

We studied the interaction of LPS with albumin, hemoglobin or high-density lipoprotein (HDL), and whether the interaction affected the activity of LPS on neutrophils. These proteins disaggregated LPS, depending upon temperature and LPS:protein ratio. Albumin-treated LPS was absorbed by immobilized anti-albumin antibody and was eluted with Triton X-100, indicating that LPS formed a hydrophobic complex with albumin. Rd mutant LPS was not disaggregated by the proteins, and did not form a complex with the proteins. But triethylamine-treated Rd mutant LPS formed complexes. When LPS was incubated with an equal concentration of albumin and with polymyxin B (PMXB), PMXB-LPS-protein three-way complexes were formed. After removal of PMXB, the complexes consisted of 11-15 LPS monomers bound to one albumin or hemoglobin molecule. LPS primed neutrophils for enhanced release of formyl peptide-stimulated superoxide, in a serum- and LPS-binding protein (LBP)-dependent manner. Although LPS plus LBP alone did not prime neutrophils, albumin-, hemoglobin- or HDL-treated LPS primed neutrophils when added with LBP. Triethylamine-treated Rd mutant LPS primed neutrophils only when incubated with one of the proteins and with LBP. Thus, in addition to LBP, disaggregation and complex formation of LPS with one of these proteins is required for LPS to prime neutrophils.

DOI： 10.1093/femspd/ftw003

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

DOI： 10.1002/iid3.99

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

Periodontal ligament stem cells (PDLSCs) are known to play a pivotal role in regenerating the periodontium. Amelogenin, which belongs to a family of extracellular matrix (ECM) proteins, is a potential bioactive molecule for periodontal regenerative therapy. However, its downstream target molecules and/or signaling patterns are still unknown. Our recent proteomic study identified glucose-regulated protein 78 (Grp78) as a new amelogenin-binding protein. In this study, we demonstrate, for the first time, the cellular responses induced by the biological interaction between amelogenin and Grp78 in the human undifferentiated PDL cell line 1-17, which possesses the most typical characteristics of PDLSCs. Confocal co-localization experiments revealed the internalization of recombinant amelogenin (rM180) via binding to cell surface Grp78, and the endocytosis was inhibited by the silencing of Grp78 in 1-17 cells. Microarray analysis indicated that rM180 and Grp78 regulate the expression profiles of cell migration-associated genes in 1-17 cells. Moreover, Grp78 overexpression enhanced rM180-induced cell migration and adhesion without affecting cell proliferation, while silencing of Grp78 diminished these activities. Finally, binding of rM180 to Grp78 promoted the formation of lamellipodia, and the simultaneous activation of Rac1 was also demonstrated by NSC23766, a widely accepted Rac1 inhibitor. These results suggest that Grp78 is essential for enhancing amelogenin-induced migration in 1-17 cells. The biological interaction of amelogenin with Grp78 offers significant therapeutic potential for understanding the biological components and specific functions involved in the signal transduction of amelogenin-induced periodontal tissue regeneration. J. Cell. Physiol. 231: 414-427, 2016.

DOI： 10.1002/jcp.25087

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

Objectives: We previously demonstrated that a dominant-negative Sprouty2 (Spry2) mutation promotes osteoblast proliferation and differentiation after basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) stimulation, whereas it diminishes proliferation of gingival epithelial cells, thereby inducing favourable conditions for periodontal tissue regeneration. In this study, we investigated how Spry2 inhibition affects the cellular physiology of periodontal ligament (PDL) cells. Methods: A total of 1-17 PDL cells (multipotent clonal human PDL cell line) were stimulated with bFGF and EGF after transfection of Spry2 siRNA. Cell proliferation, migration, ALP staining, real-time PCR, Western blot and immunofluorescence assays were performed. Results: ERK1/2 activation and proliferation of 1-17 PDL cells were significantly upregulated by the addition of Spry2 siRNA in the presence of bFGF and EGF. In addition, Spry2 siRNA reduced transcription of osteogenesis-related genes and ALP staining relative to control cells. Furthermore, it increased AKT/phosphatidylinositol 3-kinase (PI3K) phosphorylation; consequently, Rac1 but not Cdc42 was activated, thereby promoting lamellipodia formation, cell proliferation and migration after stimulation by bFGF and EGF. Conclusion: Spry2 combined with bFGF and EGF stimulation reduced PDL cell migration and proliferation with inducing osteoblastic differentiation. These in vitro findings may provide a molecular basis for novel therapeutic approaches for establishing periodontal tissue regeneration.

DOI： 10.1111/odi.12369

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

Emdogain (enamel matrix derivative, EMD) is well recognized in periodontology. It is used in periodontal surgery to regenerate cementum, periodontal ligament, and alveolar bone. However, the precise molecular mechanisms underlying periodontal regeneration are still unclear. In this study, we investigated the proteins bound to amelogenin, which are suggested to play a pivotal role in promoting periodontal tissue regeneration. To identify new molecules that interact with amelogenin and are involved in osteoblast activation, we employed coupling affinity chromatography with proteomic analysis in fractionated SaOS-2 osteoblastic cell lysate. In SaOS-2 cells, many of the amelogenin-interacting proteins in the cytoplasm were mainly cytoskeletal proteins and several chaperone molecules of heat shock protein 70 (HSP70) family. On the other hand, the proteomic profiles of amelogenin-interacting proteins in the membrane fraction of the cell extracts were quite different from those of the cytosolic-fraction. They were mainly endoplasmic reticulum (ER)-associated proteins, with lesser quantities of mitochondrial proteins and nucleoprotein. Among the identified amelogenin-interacting proteins, we validated the biological interaction of amelogenin with glucose-regulated protein 78 (Grp78/Bip), which was identified in both cytosolic and membrane-enriched fractions. Confocal co-localization experiment strongly suggested that Grp78/Bip could be an amelogenin receptor candidate. Further biological evaluations were examined by Grp78/Bip knockdown analysis with and without amelogenin. Within the limits of the present study, the interaction of amelogenin with Grp78/Bip contributed to cell proliferation, rather than correlate with the osteogenic differentiation in SaOS-2 cells. Although the biological significance of other interactions are not yet explored, these findings suggest that the differential effects of amelogenin-derived osteoblast activation could be of potential clinical significance for understanding the cellular and molecular bases of amelogenin-induced periodontal tissue regeneration.

DOI： 10.1371/journal.pone.0078129

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

Cleft palate is one of the most common craniofacial deformities. The fibroblast growth factor (FGF) plays a central role in reciprocal interactions between adjacent tissues during palatal development, and the FGF signaling pathway has been shown to be inhibited by members of the Sprouty protein family. In this study, we report the incidence of cleft palate, possibly caused by failure of palatal shelf elevation, in Sprouty2-deficient (KO) mice. Sprouty2-deficient palates fused completely in palatal organ culture. However, palate mesenchymal cell proliferation estimated by Ki-67 staining was increased in Sprouty2 KO mice compared with WT mice. Sprouty2-null palates expressed higher levels of FGF target genes, such as Msx1, Etv5, and Ptx1 than WT controls. Furthermore, proliferation and the extracellular signal-regulated kinase (Erk) activation in response to FGF was enhanced in palate mesenchymal cells transfected with Sprouty2 small interfering RNA. These results suggest that Sprouty2 regulates palate mesenchymal cell proliferation via FGF signaling and is involved in palatal shelf elevation.

DOI： 10.1016/j.bbrc.2010.12.116

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

Mouse CD1d-restricted Vα14 NKT cells are a unique subset of lymphocytes, which play important roles in immune regulation, tumor surveillance and host defense against pathogens. DOCK2, a mammalian homolog of Caenorhabditis elegans CED-5 and Drosophila melanogaster myoblast city, is critical for lymphocyte migration and regulates T cell responsiveness through immunological synapse formation, yet its role in Vα14 NKT cells remains unknown. We found that DOCK2 deficiency causes marked reduction of Vα14 NKT cells in the thymus, liver, and spleen. When α-galactesylceramide (α-GalCer), a ligand for Vα14 NKT cells, was administrated, cytokine production was scarcely detected in DOCK2-deficient mice, suggesting that DOCK2 deficiency primarily affects generation of Vα14 NKT cells. Supporting this idea, staining with CD1d/α-GalCer tetramers revealed that CD44 ^-NK1.1^- Vα14 NKT cell precursors are severely reduced in the thymuses of DOCK2-deficient mice. In addition, studies using bone marrow chimeras indicated that development of Vα14 NKT cells requires DOCK2 expression in T cell precursors, but not in APCs. These results indicate that DOCK2 is required for positive selection of Vα14 NKT cells in a cell-autonomous manner, thereby suggesting that avidity-based selection also governs development of this unique subset of lymphocytes in the thymus.

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

Allograft rejection is induced by graft tissue infiltration of alloreactive T cells that are activated mainly in secondary lymphoid organs of the host. DOCK2 plays a critical role in lymphocyte homing and immunological synapse formation by regulating the actin cytoskeleton, yet its role in the in vivo immune response remains unknown. We show here that DOCK2 deficiency enables long-term survival of cardiac allografts across a complete mismatch of the major histocompatibility complex molecules. In DOCK2-deficient mice, alloreactivity and allocytotoxicity were suppressed significantly even after in vivo priming with alloantigens, which resulted in reduced intragraft expression of effector molecules, such as interferon-γ, granzyme B, and perforin. This is mediated, at least in part, by preventing potentially alloreactive T cells from recruiting into secondary lymphoid organs. In addition, we found that DOCK2 is critical for CD28-mediated Rac activation and is required for the full activation of alloreactive T cells. Although DOCK2-deficient, alloreactive T cells were activated in vitro in the presence of exogenous interleukin-2, these T cells, when transferred adoptively, failed to infiltrate into the allografts that were transplanted into RAG1-deficient mice. Thus, DOCK2 deficiency attenuates allograft rejection by simultaneously suppressing multiple and key processes. We propose that DOCK2 could be a novel molecular target for controlling transplant rejection. JEM

DOI： 10.1084/jem.20050911

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

Clearance of apoptotic cells by macrophages is considered important for prevention of inflammatory responses leading to tissue damage. The phosphatidylserine receptor (PSR), which specifically binds to phosphatidylserine (PS) exposed on the surface of apoptotic cells, mediates uptake of apoptotic cells in vitro, yet the physiologic relevance of PSR remains unknown. This issue was addressed by generating PSR-deficient (PSR ^-/-) mice. PSR^-/- mice exhibited severe anemia and died during the perinatal period. In the PSR^-/- fetal livers, erythroid differentiation was blocked at an early erythroblast stage. In addition, PSR^-/- embryos exhibited thymus atrophy owing to a developmental defect of T-lymphoid cells. Clearance of apoptotic cells by macrophages was impaired in both liver and thymus of PSR^-/- embryos. However, this did not induce up-regulation of inflammatory cytokines. These results indicate that during embryonic development, PSR-mediated apoptotic cell uptake is required for definitive erythropoiesis and T lymphopoiesis, independently of the prevention of inflammatory responses.

DOI： 10.1182/blood-2003-09-3245

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

Organ-specific autoimmune diseases have been postulated to be the result of T cell response against organ-specific self-peptides bound to MHC molecules. Contrary to this paradigm, we report here that transgenic mice lacking MHC class I expression and expressing an MHC class II I-A^b molecule that presents only a single peptide (Eα52-68) spontaneously develops peripheral nervous system-specific autoimmune disease with many of the histopathological features found in experimental allergic neuritis. Reciprocal bone marrow chimeras produced using susceptible and resistant lines revealed that bone marrow-derived cells determined disease susceptibility. While the expression of the I-A^b-Eα52-68 complex in the periphery was readily detectable in both lines, its expression on thymic dendritic cells responsible for tolerance induction was markedly lower in the susceptible line than in the resistant line. Consistent with this, CD4⁺ T cells that can be activated by the I-A^b-Eα52-68 complex were found in the susceptible line, but not in the resistant line. Such CD4⁺ T cells conferred the disease to the resistant line by adoptive transfer, and administration of Ab specific for the I-A^b-Eα52-68 complex inhibited disease manifestation in the susceptible line. These results indicate that disease development involves systemic T cell reactivity to I-A^b-Eα52-68 complex, probably caused by incomplete negative thymocyte selection.

DOI： 10.1172/JCI200113256

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

Cell migration is a fundamental biological process involving membrane polarization and cytoskeletal dynamics, both of which are regulated by Rho family GTPases. Among these molecules, Rac is crucial for generating the actin-rich lamellipodial protrusion, a principal part of the driving force for movement. The CDM family proteins, Caenorhabditis elegans CED-5, human DOCK180 and Drosophila melanogaster Myoblast City (MBC), are implicated to mediate membrane extension by functioning upstream of Rac. Although genetic analysis has shown that CED-5 and Myoblast City are crucial for migration of particular types of cells, physiological relevance of the CDM family proteins in mammals remains unknown. Here we show that DOCK2, a haematopoietic cell-specific CDM family protein, is indispensable for lymphocyte chemotaxis. DOCK2-deficient mice (DOCK2^-/-) exhibited migration defects of T and B lymphocytes, but not of monocytes, in response to chemokines, resulting in several abnormalities including T lymphocytopenia, atrophy of lymphoid follicles and loss of marginal-zone B cells. In DOCK2^-/- lymphocytes, chemokine-induced Rac activation and actin poly- merization were almost totally abolished. Thus, in lymphocyte migration DOCK2 functions as a central regulator that mediates cytoskeletal reorganization through Rac activation.

DOI： 10.1038/35090591

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

T cell differentiation in the thymus is driven by positive selection through the interaction of αβ T cell receptors (TCRs) with selfpeptides bound to self-major histocompatibility complex molecules, yet the influence of the peptide sequence on this process remains unknown. To address this issue, we have compared CD4⁺ T cell differentiation between two sets of mouse lines in which MHC class II I-A^b molecules are occupied with either Eα chain-derived peptide (pEα) or its variant, (p)60k, with one amino acid substitution from leucine to lysine at P5 residue of TCR contacts. Here, we show that despite the comparable expression of I-A^b-peptide complex in the thymus, this substitution from leucine to lysine affects efficiency of positive selection, resulting in extremely small numbers of CD4⁺ T cells to be selected to mature on I-A^b-(p)60k complex. Furthermore, we show that, although I-A^b-pEα complex selects diverse T cells, T cell repertoire shaped by I-A^b-(p)60k complex is markedly constrained. Our findings thus suggest that positive selection is both specific and degenerate, depending on the amino acid residues at TCR contacts of the selecting self-peptides.

DOI： 10.1073/pnas.250470797

Responsible for pages：41巻3号,243-247頁   Language：Japanese   Book type：General book, introductory book for general audience

Responsible for pages：22巻11号,1204-1207頁   Language：Japanese   Book type：General book, introductory book for general audience

Responsible for pages：医薬の門社   Language：Japanese   Book type：General book, introductory book for general audience

Responsible for pages：33巻1号,52-54頁   Language：Japanese   Book type：General book, introductory book for general audience

Responsible for pages：p102-106   Language：Japanese   Book type：Scholarly book

Language：Japanese   Book type：Scholarly book

Language：Japanese   Book type：Scholarly book

Event date： 2015.3

Language：English   Presentation type：Oral presentation (general)  

Venue：Haynes Convention Center, Boston   Country：United States  

Event date： 2015.3

Language：English   Presentation type：Oral presentation (general)  

Venue：Haynes Convention Center, Boston   Country：United States  

Event date： 2014.12

Language：English   Presentation type：Oral presentation (general)  

Venue：KKRホテル大阪、大阪市   Country：Japan  

Event date： 2013.9

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：前橋市   Country：Japan  

Event date： 2013.9

Language：English   Presentation type：Oral presentation (general)  

Venue：Nara   Country：Japan  

Event date： 2013.8

Language：English   Presentation type：Oral presentation (general)  

Venue：Plaza Athenee, Bangkok   Country：Thailand  

Event date： 2013.3

Language：English   Presentation type：Oral presentation (general)  

Venue：Washington States Convention Center   Country：United States  

Event date： 2013.3

Presentation type：Symposium, workshop panel (public)  

Venue：Fukuoka Recent Hotel   Country：Japan  

Periodontal disease is a chronic inflammatory response to bacterial pathogens involving the supporting tissues of the teeth. The anerobic bacterium, Porphyromonas gingivalis, has been implicated as a major etiologic agent in the development and progression of periodontitis. Lipopolysaccharide (LPS) from P. gingivalis induces pro-inflammatory cytokines in host macrophages. Macrophages can acquire distinct functional phenotypes, referred to as classically activated, pro-inflammatory macrophages (M1) and alternatively activated, anti-inflammatory macrophages (M2). M2 macrophages are associated with homeostatic functions linked to wound healing and tissue repair. Therefore, the inhibition of pro-inflammatory cytokine production in macrophages is the important biological reaction for the homeostasis after elimination of the etiologic agent.
Sprouty proteins are identified as inhibitors of fibroblast growth factor (FGF) receptor. Specifically Sprouty2 (Spry2) functions as a negative regulator of receptor tyrosine kinases (RTKs) signaling, and is expressed in several developing organs including kidney, lung, brain, heart, skeletal muscle and craniofacial area. Furthermore, Spry2 has conserved function to modulate morphogenesis in several tissues. Recently, we demonstrated that inhibition of Spry2 induced cell proliferation and differentiation of MC3T3-E1osteoblastic cells, while it diminished cell proliferation of GE1gingival
epithelial cells in vitro.
We report here that suppression of Spry2 polarizes J774 mouse macrophages toward alternative macrophage activation (M2) phenotype when J774 cells are stimulated with LPS from P. gingivalis.
This polarization is defined by surface protein expression, cytokine production and arginase activity. These data provide evidence that Spry2 inhibitors affect the inflammatory process at the level of the
macrophage and shifts macrophage polarization from pro-inflammatory properties toward anti-inflammatory ones in periodontitis.
Additionally, we are confirming how this response affects the periodontal tissue regeneration or the para-inflammation of periodontitis.

Event date： 2012.11

Venue：大阪国際会議場   Country：Japan  

Event date： 2012.10 - 2012.9

Venue：Los Angels Convention Center   Country：United States  

Event date： 2012.9

Venue：Finlandia Hall   Country：Finland  

Event date： 2010.7

Presentation type：Oral presentation (general)  

Venue：Center Conventions International Barcelona (CCIB)   Country：Spain  

Event date： 2009.4

Venue：Miami Beach Convention Center   Country：United States  

Analysis of Streptococcus mutans Biofilm Proteins Recognized by Salivary IgA.

Presentation type：Oral presentation (general)  

Venue：第32回日本免疫学会総会・学術集会　東京   Country：Japan  

Venue：第30回日本免疫学会総会・学術集会　仙台   Country：Japan  

Presentation type：Oral presentation (general)  

Country：Japan  

Event date： 2024.3

Language：English  

Venue：New Orleans   Country：United States  

Event date： 2024.3

Language：English  

Venue：New Orleans   Country：United States  

Event date： 2023.12

Language：Japanese  

Venue：アバンセホール、 佐賀市   Country：Japan  

Event date： 2023.10

Language：Japanese  

Venue：出島メッセ長崎、 長崎市   Country：Japan  

Event date： 2023.6

Language：Japanese  

Venue：くにびきメッセ、松江市、島根県   Country：Japan  

Event date： 2022.11

Language：Japanese  

Venue：Web開催   Country：Japan  

Event date： 2022.11

Language：Japanese  

Venue：Web開催   Country：Japan  

Event date： 2022.10

Language：English  

Venue：Phoenix, AZ   Country：United States  

Event date： 2022.10

Language：English  

Venue：Phoenix, AZ   Country：United States  

Event date： 2022.10

Language：English  

Venue：Phoenix, AZ   Country：United States  

Event date： 2022.6

Language：English  

Venue：WEB開催   Country：China  

Event date： 2022.6

Language：Japanese  

Venue：Web開催   Country：Japan  

Event date： 2022.6

Language：Japanese  

Venue：Web開催   Country：Japan  

Event date： 2022.6

Language：Japanese  

Venue：東京京王プラザホテル   Country：Japan  

Event date： 2022.6

Language：Japanese  

Venue：東京京王プラザホテル   Country：Japan  

Event date： 2021.11

Language：English  

Venue：九州大学歯学部   Country：Japan  

Event date： 2021.11

Language：English  

Venue：九州大学歯学部   Country：Japan  

Event date： 2021.10 - 2021.11

Language：Japanese  

Venue：Web開催   Country：Japan  

Event date： 2021.10

Language：English  

Venue：九州大学医学部百年講堂   Country：Japan  

Event date： 2021.10

Language：English  

Venue：九州大学医学部百年講堂   Country：Japan  

Event date： 2021.10

Language：English  

Venue：九州大学医学部百年講堂   Country：Japan  

Event date： 2021.6

Language：Japanese  

Venue：Web開催   Country：Japan  

Event date： 2021.5 - 2021.6

Language：Japanese  

Venue：Web開催   Country：Japan  

Event date： 2020.11

Language：Japanese  

Venue：Web開催   Country：Japan  

Event date： 2020.11

Language：Japanese  

Venue：Web開催   Country：Japan  

Event date： 2020.11

Language：English  

Venue：WEB開催   Country：Japan  

Event date： 2020.11

Language：English  

Venue：WEB開催   Country：Japan  

Event date： 2020.11

Language：English  

Venue：WEB開催   Country：United States  

Event date： 2020.11

Language：English  

Venue：WEB開催   Country：United States  

Event date： 2020.11

Language：English  

Venue：WEB開催   Country：United States  

Event date： 2020.10

Language：Japanese  

Venue：Web開催   Country：Japan  

Event date： 2020.3

Language：English  

Venue：Washington D.C., USA（誌上開催）   Country：United States  

Event date： 2020.3

Language：English  

Venue：Washington D.C., USA（誌上開催）   Country：United States  

Event date： 2019.11

Language：Japanese  

Venue：アクロス福岡、福岡市   Country：Japan  

Event date： 2019.11

Language：Japanese  

Venue：アクロス福岡、福岡市   Country：Japan  

Event date： 2019.11

Language：Japanese  

Venue：福岡国際会議場、福岡市   Country：Japan  

Event date： 2019.10 - 2019.6

Language：Japanese  

Venue：西日本総合展示場・北九州国際会議場、北九州市   Country：Japan  

Event date： 2019.6

Language：Japanese  

Venue：石川県立音楽堂、金沢市   Country：Japan  

Event date： 2019.6

Language：Japanese  

Venue：石川県立音楽堂、金沢市   Country：Japan  

Event date： 2019.5

Language：Japanese  

Venue：神奈川県民ホール、横浜市   Country：Japan  

Event date： 2019.5

Language：English  

Venue：神奈川県民ホール、横浜市   Country：Japan  

Event date： 2019.3

Language：English  

Venue：Fukuoka   Country：Japan  

Event date： 2019.3

Language：English  

Venue：Seoul, South Korea   Country：Korea, Republic of  

Event date： 2018.11

Language：Japanese  

Venue：アクロス福岡、福岡市   Country：Japan  

Event date： 2018.11

Language：English  

Venue：みやこめっせ、京都市   Country：Japan  

Event date： 2018.7

Language：Japanese  

Venue：リーガロイヤルホテル大阪、大阪市   Country：Japan  

Event date： 2018.7

Language：English  

Venue：London   Country：United Kingdom  

Event date： 2018.6

Language：Japanese  

Venue：横浜みなとみらいホール、横浜市   Country：Japan  

Event date： 2018.6

Language：Japanese  

Venue：京王プラザホテル、東京都   Country：Japan  

Event date： 2017.12

Language：Japanese  

Venue：国立京都国際会館、京都市   Country：Japan  

Event date： 2017.12

Language：English  

Venue：国立京都国際会館、京都市   Country：Japan  

Event date： 2017.6

Language：English  

Venue：University of Pennsylvania, Philadelphia, PA, USA   Country：Japan  

Event date： 2017.6

Language：Japanese  

Venue：リンクステーションホール青森　青森市   Country：Japan  

Event date： 2016.11

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：長崎県歯科医師会館、長崎市   Country：Japan  

Event date： 2016.10

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：キッセイ文化ホール、松本市   Country：Japan  

Event date： 2016.10

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：キッセイ文化ホール、松本市   Country：Japan  

Event date： 2016.10

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：朱鷺メッセ 新潟コンベンションセンター、新潟市   Country：Japan  

Event date： 2016.6 - 2018.6

Language：English  

Venue：Seoul, South Korea   Country：Japan  

Event date： 2016.6 - 2018.6

Language：English  

Venue：Seoul, South Korea   Country：Japan  

Event date： 2016.6 - 2018.6

Language：English  

Venue：Seoul, South Korea   Country：Japan  

Event date： 2016.6

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：栃木県総合文化センター   Country：Japan  

Event date： 2016.5

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：かごしま県民交流センター、鹿児島市   Country：Japan  

Event date： 2016.2 - 2018.2

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

Venue：福岡リーセントホテル   Country：Japan  

Event date： 2016.2

Language：English   Presentation type：Oral presentation (general)  

Venue：Penn Dental Medicine in Philadelphia, Pennsylvania   Country：United States  

Event date： 2015.11

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：文京シビックホール、東京都   Country：Japan  

Event date： 2015.11

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：文京シビックホール、東京都   Country：Japan  

Event date： 2015.11

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：文京シビックホール、東京都   Country：Japan  

Event date： 2015.11

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：アクロス福岡、福岡市   Country：Japan  

Event date： 2015.10

Language：English   Presentation type：Oral presentation (general)  

Venue：福岡国際会議場   Country：Japan  

Event date： 2015.6 - 2015.7

Language：English   Presentation type：Oral presentation (general)  

Venue：Penn Dental Medicine in Philadelphia, Pennsylvania   Country：United States  

Event date： 2015.6 - 2015.7

Language：English   Presentation type：Oral presentation (general)  

Venue：Penn Dental Medicine in Philadelphia, Pennsylvania   Country：United States  

Event date： 2015.6

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：北九州国際会議場、北九州市   Country：Japan  

Event date： 2015.6

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：北九州国際会議場、北九州市   Country：Japan  

Event date： 2015.6

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：北九州国際会議場、北九州市   Country：Japan  

Event date： 2015.6

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：北九州国際会議、北九州市   Country：Japan  

Event date： 2015.5

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：幕張メッセ 国際会議場、千葉市   Country：Japan  

Event date： 2015.3

Language：English   Presentation type：Oral presentation (general)  

Venue：Haynes Convention Center, Boston   Country：United States  

Event date： 2015.2

Language：English   Presentation type：Oral presentation (general)  

Venue：福岡リーセントホテル、福岡市   Country：Japan  

Event date： 2015.2

Language：English   Presentation type：Oral presentation (general)  

Venue：福岡リーセントホテル、福岡市   Country：Japan  

Event date： 2014.12

Language：English   Presentation type：Oral presentation (general)  

Venue：KKRホテル大阪、大阪市   Country：Japan  

Event date： 2014.11

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：アクロス福岡、福岡市   Country：Japan  

Event date： 2014.11

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：アクロス福岡、福岡市   Country：Japan  

Event date： 2014.10

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：山形テルサ、山形市   Country：Japan  

Event date： 2014.10

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：神戸市国際展示場、神戸市   Country：Japan  

Event date： 2014.10

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：神戸市国際展示場、神戸市   Country：Japan  

Event date： 2014.6

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：滋賀県立劇場びわ湖ホール、大津市   Country：Japan  

Event date： 2014.6

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：滋賀県立劇場びわ湖ホール、大津市   Country：Japan  

Event date： 2014.5

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：長良川国際会議場、岐阜市   Country：Japan  

Event date： 2013.11

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：秋田市   Country：Japan  

Event date： 2013.11

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：北九州市九州歯科大学   Country：Japan  

Event date： 2013.9

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：前橋市   Country：Japan  

Event date： 2013.9

Language：English   Presentation type：Oral presentation (general)  

Venue：Nara   Country：Japan  

Event date： 2013.9

Language：English   Presentation type：Oral presentation (general)  

Venue：Nara   Country：Japan  

Event date： 2013.9

Language：English   Presentation type：Oral presentation (general)  

Venue：Nara   Country：Japan  

Event date： 2013.8

Language：English   Presentation type：Oral presentation (general)  

Venue：Plaza Athenee, Bangkok   Country：Thailand  

Event date： 2013.8

Language：English   Presentation type：Oral presentation (general)  

Venue：Plaza Athenee, Bangkok   Country：Thailand  

Event date： 2013.8

Language：English   Presentation type：Oral presentation (general)  

Venue：Plaza Athenee, Bangkok   Country：Thailand  

Event date： 2013.5 - 2013.6

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：タワーホール船堀   Country：Japan  

Event date： 2013.3

Language：English   Presentation type：Oral presentation (general)  

Venue：Washington States Convention Center   Country：United States  

Event date： 2013.3

Language：English   Presentation type：Oral presentation (general)  

Venue：Washington States Convention Center   Country：United States  

Event date： 2013.3

Presentation type：Oral presentation (general)  

Venue：Fukuoka Recent Hotel   Country：Japan  

Event date： 2013.3

Presentation type：Oral presentation (general)  

Venue：Fukuoka Recent Hotel   Country：Japan  

Event date： 2012.11

Venue：大阪国際会議場   Country：Japan  

Event date： 2012.11

Language：Japanese  

Venue：大阪国際会議場   Country：Japan  

Event date： 2012.9 - 2012.10

Country：United States  

Event date： 2012.9 - 2012.11

Venue：Finlandia Hall   Country：Finland  

Event date： 2012.9 - 2012.11

Venue：Finlandia Hall   Country：Finland  

Event date： 2012.9

Venue：Finlandia Hall   Country：Finland  

Event date： 2012.9

Venue：Finlandia Hall   Country：Finland  

Event date： 2012.5

Presentation type：Oral presentation (general)  

Venue：札幌コンベンションセンター   Country：Japan  

Event date： 2011.11

Presentation type：Oral presentation (general)  

Venue：長崎県歯科医師会館   Country：Japan  

Event date： 2011.9

Presentation type：Oral presentation (general)  

Venue：海峡メッセ下関   Country：Japan  

Event date： 2011.3

Presentation type：Oral presentation (general)  

Venue：San Diego Convention Center   Country：United States  

Event date： 2010.10 - 2010.11

Presentation type：Oral presentation (general)  

Venue：Hawaii Convention Center   Country：United States  

Event date： 2010.7

Presentation type：Oral presentation (general)  

Venue：Center Conventions International Barcelona (CCIB)   Country：Spain  

Event date： 2010.7

Presentation type：Oral presentation (general)  

Venue：Center Conventions International Barcelona (CCIB)   Country：Spain  

Event date： 2010.7

Presentation type：Oral presentation (general)  

Venue：Center Conventions International Barcelona (CCIB)   Country：Spain  

Event date： 2010.2

Presentation type：Oral presentation (general)  

Venue：九州大学医学部 百年講堂   Country：Japan  

Event date： 2009.11

Presentation type：Oral presentation (general)  

Venue：Ramada Plaza, Jeju   Country：Korea, Republic of  

Event date： 2009.5

Venue：岡山コンベンションセンター   Country：Japan  

Analysis of the nobel protein bound to amelogenin in periodontal tissue cells.

Venue：第33回日本免疫学会総会・学術集会　福岡   Country：Japan  

Venue：第29回日本免疫学会総会・学術集会　京都   Country：Japan  

Venue：第29回日本免疫学会総会・学術集会　京都   Country：Japan  

Venue：第30回日本免疫学会総会・学術集会　仙台   Country：Japan  

Venue：第30回日本免疫学会総会・学術集会　仙台   Country：Japan  

Venue：第31回日本免疫学会総会・学術集会　大阪   Country：Japan  

Language：Japanese  

Language：Japanese  

Language：English  

Language：English  

Language：Japanese  

Language：Japanese  

Language：Japanese  

Language：Japanese  

Language：Japanese  

Language：Japanese  

Language：English  

Language：English  

Language：English   Publishing type：Article, review, commentary, editorial, etc. (scientific journal)  

DOI： doi: 10.1111/prd.12298.

Language：English   Publishing type：Article, review, commentary, editorial, etc. (scientific journal)  

DOI： doi:10.4172/2168-9431.1000137

Language：English   Publishing type：Article, review, commentary, editorial, etc. (scientific journal)  

DOI： doi:10.4172/jcs.1000118.

Type：Competition, symposium, etc. 

Number of participants：50

Type：Peer review 

Number of peer-reviewed articles in foreign language journals：1

Type：Competition, symposium, etc. 

Type：Peer review 

Number of peer-reviewed articles in foreign language journals：1

Type：Competition, symposium, etc. 

Number of participants：2,800

Type：Peer review 

Number of peer-reviewed articles in foreign language journals：3

Type：Competition, symposium, etc. 

Type：Academic society, research group, etc. 

Type：Academic society, research group, etc. 

Type：Competition, symposium, etc. 

Type：Competition, symposium, etc. 

OSCE タイムキーパー責任者

OSCE タイムキーパー責任者

OSCE タイムキーパー責任者

Advanced OSCE実施責任者

OSCE タイムキーパー責任者

OSCE タイムキーパー責任者

Advanced OSCE実施責任者

2015年度OSCE実行委員会副委員長補佐実施責任者

Advanced OSCE実施責任者

Advanced OSCE評価者

2014年度OSCE実行委員会副委員長補佐実施責任者

Advanced OSCE評価者

平成25年度CBT問題作成ワーキンググループC責任者

OSCE実行委員会委員・ステーション４内部評価者

OSCE実行委員会委員・ステーション４内部評価者

平成24年度CBT問題作成ワーキンググループC責任者

平成23年度九州大学病院歯科医師臨床研修指導式講習会

九州大学歯学研究院 FD「カリキュラムプランニングワークショップ」参加

OSCE実行委員会委員・ステーション２総責任者

平成23年度第2回共用試験歯学系ＯＳＣＥ評価者養成ワークショップⅠ

平成23年度CBT問題作成

OSCE実行委員会補助

平成22年度CBT問題作成

間違い探し問題作成説明会

OSCE実行委員会評価者

第3回医療系大学e-ラーニング全国交流会

間違い探しを基盤とする洞察力育成医療教育

Audience：General,　Scientific,　Company,　Civic organization,　Governmental agency

Type：Other

Audience：General,　Scientific,　Company,　Civic organization,　Governmental agency

Type：Lecture

Audience：General,　Scientific,　Company,　Civic organization,　Governmental agency

Type：Lecture