Authorship：Lead author,　Last author,　Corresponding author   Language：English   Publisher：Frontiers in Immunology  

Introduction: Amelogenin, used as a periodontal tissue regeneration material, promotes healing after periodontal surgery. A previous study has demonstrated that amelogenin is taken up by macrophages into the nucleus and inhibits major histocompatibility class II (MHC II) expression at the transcriptional level, thereby suppressing subsequent T cell activation. Therefore, in this study, we focused on the suppressive effect of amelogenin on MHC II expression and examined the effect of amelogenin on graft rejection following allogeneic skin transplantation in mice with different MHC II haplotype antigens. Methods and results: Skin grafts were treated with recombinant murine amelogenin (rM180) and transplanted into recipient mice. The rM180-treated group showed a significant increase in graft survival for up to 5.5 days and a lower necrotic score than the control group. Inflammatory cell infiltration and MHC II⁺ cells were significantly lower in the rM180 group. Furthermore, serum interferon-γ, interleukin-2, and interleukin-17A levels, splenic T-helper type 1 cells and helper type 17/regulatory T cells balance were reduced in the rM180 group. RNA sequencing analysis suggested "negative regulation of immune response" and "regeneration of myocytes and myofibrils" by amelogenin treatment. Among the upregulated genes in the rM180 group, “POU domain class 2 transcription factor 2,” “lipocalin 2,” and “chitinase-like 4” were ranked high. Additionally, the ratio of M2 macrophages significantly increased in rM180-treated grafts. Discussion: These results may suggest that amelogenin can be a safe immunosuppressant or therapeutic agent against autoimmune diseases without inducing unfavorable side effects.

DOI： 10.3389/fimmu.2025.1663437

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Language：English   Publisher：Elsevier  

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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CiNii Research

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.

DOI： 10.1155/2024/8864513

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CiNii Research

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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Language：Japanese   Publisher：日本総合歯科学会  

定期的な口腔内清掃を希望する患者が来院するが,歯周疾患の治癒・病状安定を維持している患者が多い一方,病状が進行し予後不良の歯が出現しているものの,様々な理由で治療や抜歯を希望しない患者もいる。今回,患者情報を参考に,様々なコミュニケーション技法を応用して患者と治療方針を検討した。その結果,抜歯を含めた全顎的な治療に取り組むことができた症例を経験したため報告する。症例は76歳,男性。「右上の歯が痛い。」という主訴で来院した。患者は九州大学病院口腔総合診療科で全顎的治療を行った後にSupportive Periodontal Therapy(SPT)へ移行したが,その後約5年にわたり「簡単に掃除をやってほしい。」との訴えが続いた。そのため,定期的な口内法X線写真撮影や歯周基本検査を含む,十分なSPTを行えていなかった。そこで,過去の患者情報から患者の考え方を考察し,(1)目標設定理論を応用し,何か処置をする前には,処置内容,所要時間,その処置が必要な理由をその都度伝えること,(2)"But You Are Free"(BYAF) compliance-gaining techniqueを応用した質問形式を取ること,(3)"Self Persuasion"自主説得理論を応用した治療の提案を行うことのコミュニケーション技法を応用して治療を開始した。その結果,患者は現症や治療方法を十分理解・納得し,予後不良と判断した歯の抜歯と,上顎即時義歯を製作することができた。以上のことから,患者情報や様々なコミュニケーション技法を応用することで,治療へ移行ができる可能性があると考えられた。(著者抄録)

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Language：Japanese  

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Language：Japanese