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DOI： 10.1016/j.jds.2024.02.025

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DOI： 10.1016/j.jds.2023.10.022

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DOI： 10.1016/j.jds.2023.11.019

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Language：English  

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Authorship：Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：Elsevier BV  

DOI： 10.1016/j.jds.2024.05.028

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Publishing type：Research paper (scientific journal)   Publisher：MDPI AG  

Periodontal ligament-associated protein 1 (PLAP-1), also known as Asporin, is an extracellular matrix protein expressed in the periodontal ligament and plays a crucial role in periodontal tissue homeostasis. Our previous research demonstrated that PLAP-1 may inhibit TLR2/4-mediated inflammatory responses, thereby exerting a protective function against periodontitis. However, the precise roles of PLAP-1 in the periodontal ligament (PDL) and its relationship to periodontitis have not been fully explored. In this study, we employed PLAP-1 knockout mice to investigate its roles and contributions to PDL tissue and function in a ligature-induced periodontitis model. Mandibular bone samples were collected from 10-week-old male C57BL/6 (WT) and PLAP-1 knockout (KO) mice. These samples were analyzed through micro-computed tomography (μCT) scanning, hematoxylin and eosin (HE) staining, picrosirius red staining, and fluorescence immunostaining using antibodies targeting extracellular matrix proteins. Additionally, the structure of the PDL collagen fibrils was examined using transmission electron microscopy (TEM). We also conducted tooth extraction and ligature-induced periodontitis models using both wild-type and PLAP-1 KO mice. PLAP-1 KO mice did not exhibit any changes in alveolar bone resorption up to the age of 10 weeks, but they did display an enlarged PDL space, as confirmed by μCT and histological analyses. Fluorescence immunostaining revealed increased expression of extracellular matrix proteins, including Col3, BGN, and DCN, in the PDL tissues of PLAP-1 KO mice. TEM analysis demonstrated an increase in collagen diameter within the PDL of PLAP-1 KO mice. In line with these findings, the maximum stress required for tooth extraction was significantly lower in PLAP-1 KO mice in the tooth extraction model compared to WT mice (13.89 N ± 1.34 and 16.51 N ± 1.31, respectively). In the ligature-induced periodontitis model, PLAP-1 knockout resulted in highly severe alveolar bone resorption, with a higher number of collagen fiber bundle tears and significantly more osteoclasts in the periodontium. Our results demonstrate that mice lacking PLAP-1/Asporin show alteration of periodontal ligament structures and acceleration of bone loss in periodontitis. This underscores the significant role of PLAP-1 in maintaining collagen fibrils in the PDL and suggests the potential of PLAP-1 as a therapeutic target for periodontal diseases.

DOI： 10.3390/ijms242115989

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DOI： 10.1016/j.jds.2023.09.027

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Abstract

Dental pulp stem cells (DPSC) usually remain quiescent in the dental pulp tissue; however, once the dental pulp tissue is injured, DPSCs potently proliferate and migrate into the injury microenvironment and contribute to immuno-modulation and tissue repair. However, the key molecules that physiologically support the potent proliferation and migration of DPSCs have not been revealed. In this study, we searched publicly available transcriptome raw data sets, which contain comparable (i.e., equivalently cultured) DPSC and mesenchymal stem cell data. Three data sets were extracted from the Gene Expression Omnibus database and then processed and analyzed. MXRA5 was identified as the predominant DPSC-enriched gene associated with the extracellular matrix. MXRA5 is detected in human dental pulp tissues. Loss of MXRA5 drastically decreases the proliferation and migration of DSPCs, concomitantly with reduced expression of the genes associated with the cell cycle and microtubules. In addition to the known full-length isoform of MXRA5, a novel splice variant of MXRA5 was cloned in DPSCs. Recombinant MXRA5 coded by the novel splice variant potently induced the haptotaxis migration of DPSCs, which was inhibited by microtubule inhibitors. Collectively, MXRA5 is a key extracellular matrix protein in dental pulp tissue for maintaining the proliferation and migration of DPSCs.

DOI： 10.1038/s41598-023-42684-z

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Other Link： https://www.nature.com/articles/s41598-023-42684-z

Authorship：Lead author,　Corresponding author   Language：Japanese   Publishing type：Research paper (scientific journal)  

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Authorship：Corresponding author   Publishing type：Research paper (scientific journal)  

DOI： 10.3390/ijms24020970

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Periodontitis is characterized by irreversible destruction of periodontal tissue. At present, the accepted etiology of periodontitis is based on a three-factor theory including pathogenic bacteria, host factors, and acquired factors. Periodontitis development usually takes a decade or longer and is therefore called chronic periodontitis (CP). To search for genetic factors associated with CP, several genome-wide association study (GWAS) analyses were conducted; however, polymorphisms associated with CP have not been identified. Epigenetics, on the other hand, involves acquired transcriptional regulatory mechanisms due to reversibly altered chromatin accessibility. Epigenetic status is a condition specific to each tissue and cell, mostly determined by the responses of host cells to stimulations by local factors, like bacterial inflammation, and systemic factors such as nutrition status, metabolic diseases, and health conditions. Significantly, epigenetic status has been linked with the onset and progression of several acquired diseases. Thus, epigenetic factors in periodontal tissues are attractive targets for periodontitis diagnosis and treatments. In this review, we introduce accumulating evidence to reveal the epigenetic background effects related to periodontitis caused by genetic factors, systemic diseases, and local environmental factors, such as smoking, and clarify the underlying mechanisms by which epigenetic alteration influences the susceptibility of periodontitis.

DOI： 10.1016/j.jdsr.2022.06.001

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Authorship：Corresponding author   Publishing type：Research paper (scientific journal)   Publisher：SAGE Publications  

DOI： 10.1177/00220345221075968

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Frontiers Media SA  

Cementum resorption, unlike bone resorption, is clinically known to occur only with limited pathological stimuli, such as trauma, orthodontic forces, and large apical periodontitis; however, the molecular mechanisms that control osteoclast formation on the cementum surface remain unclear. In this study, we focused on extracellular vesicles (EVs) secreted by cementoblasts and analyzed their effects on osteoclast differentiation. EVs were extracted from the conditioned medium (CM) of the mouse cementoblast cell line OCCM-30. Transmission electron microscopy (TEM) analysis confirmed the presence of EVs with a diameter of approximately 50–200 nm. The effect of the EVs on osteoclast differentiation was examined using the mouse osteoclast progenitor cell line RAW 264.7 with recombinant receptor activator of nuclear factor (NF)-κB ligand (rRANKL) stimulation. EVs enhanced the formation of tartrate-resistant acid phosphatase (TRAP) activity-positive cells upon rRANKL stimulation. EVs also enhanced the induction of osteoclast-associated gene and protein expression in this condition, as determined by real-time PCR and Western blotting, respectively. On the other hand, no enhancing effect of EVs was observed without rRANKL stimulation. A Western blot analysis revealed no expression of receptor activator of NF-κB ligand (RANKL) in EVs themselves. The effect on rRANKL-induced osteoclast differentiation was examined using the CM of cementoblasts in terms of TRAP activity-positive cell formation and osteoclast-associated gene expression. The conditioned medium partly inhibited rRANKL-induced osteoclast differentiation and almost completely suppressed its enhancing effect by EVs. These results indicate that cementoblasts secreted EVs, which enhanced RANKL-induced osteoclast differentiation, and simultaneously produced soluble factors that neutralized this enhancing effect of EVs, implicating this balance in the regulation of cementum absorption. A more detailed understanding of this crosstalk between cementoblasts and osteoclasts will contribute to the development of new therapies for pathological root resorption.

DOI： 10.3389/fphys.2022.825596

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Publishing type：Research paper (scientific journal)   Publisher：Public Library of Science (PLoS)  

Amelogenins, major extra cellular matrix proteins of developing tooth enamel, are predominantly expressed by ameloblasts and play significant roles in the formation of enamel. Recently, amelogenin has been detected in various epithelial and mesenchymal tissues, implicating that it might have distinct functions in various tissues. We have previously reported that leucine rich amelogenin peptide (LRAP), one of the alternate splice forms of amelogenin, regulates receptor activator of NF-kappa B ligand (RANKL) expression in cementoblast/periodontal ligament cells, suggesting that the amelogenins, especially LRAP, might function as a signaling molecule in bone metabolism. The objective of this study was to identify and define LRAP functions in bone turnover. We engineered transgenic (TgLRAP) mice using a murine 2.3kb α1(I)-collagen promoter to drive expression of a transgene consisting of LRAP, an internal ribosome entry site (IRES) and enhanced green fluorescent protein (EGFP) to study functions of LRAP in bone formation and resorption. Calvarial cell cultures from the TgLRAP mice showed increased alkaline phosphatase (ALP) activity and increased formation of mineralized nodules compared to the cells derived from wild-type (WT) mice. The TgLRAP calvarial cells also showed an inhibitory effect on osteoclastogenesis <italic>in vitro</italic>. Gene expression comparison by quantitative polymerase chain reaction (Q-PCR) in calvarial cells indicated that bone formation makers such as <italic>Runx2</italic>, <italic>Alp</italic>, and <italic>osteocalcin</italic> were increased in TgLRAP compared to the WT cells. Meanwhile, <italic>Rankl</italic> expression was decreased in the TgLRAP cells <italic>in vitro</italic>. The ovariectomized (OVX) TgLRAP mice resisted bone loss induced by ovariectomy resulting in higher bone mineral density in comparison to OVX WT mice. The quantitative analysis of calcein intakes indicated that the ovariectomy resulted in increased bone formation in both WT and TgLRAP mice; OVX TgLRAP appeared to show the most remarkably increased bone formation. The parameters for bone resorption in tissue sections showed increased number of osteoclasts in OVX WT, but not in OVX TgLRAP over that of sham operated WT or TgLRAP mice, supporting the observed bone phenotypes in OVX mice. This is the first report identifying that LRAP, one of the amelogenin splice variants, affects bone turnover <italic>in vivo</italic>.

DOI： 10.1371/journal.pone.0259966

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The periodontal ligament is a soft connective tissue embedded between the alveolar bone and cementum, the surface hard tissue of teeth. Periodontal ligament fibroblasts (PDLF) actively express osteo/cementogenic genes, which contribute to periodontal tissue homeostasis. However, the key factors maintaining the osteo/cementogenic abilities of PDLF remain unclear. We herein demonstrated that PPARγ was expressed by in vivo periodontal ligament tissue and its distribution pattern correlated with alkaline phosphate enzyme activity. The knockdown of PPARγ markedly reduced the osteo/cementogenic abilities of PDLF in vitro, whereas PPARγ agonists exerted the opposite effects. PPARγ was required to maintain the acetylation status of H3K9 and H3K27, active chromatin markers, and the supplementation of acetyl-CoA, a donor of histone acetylation, restored PPARγ knockdown-induced decreases in the osteo/cementogenic abilities of PDLF. An RNA-seq/ChIP-seq combined analysis identified four osteogenic transcripts, RUNX2, SULF2, RCAN2, and RGMA, in the PPARγ-dependent active chromatin region marked by H3K27ac. Furthermore, RUNX2-binding sites were selectively enriched in the PPARγ-dependent active chromatin region. Collectively, these results identified PPARγ as the key transcriptional factor maintaining the osteo/cementogenic abilities of PDLF and revealed that global H3K27ac modifications play a role in the comprehensive osteo/cementogenic transcriptional alterations mediated by PPARγ.

DOI： 10.3390/ijms22168646

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Publishing type：Research paper (scientific journal)   Publisher：Frontiers Media {SA}  

<jats:p>Loeys–Dietz syndrome (LDS) is a syndromic connective tissue disorder caused by a heterozygous missense mutation in genes that encode transforming growth factor (TGF)-β receptor (<jats:italic>TGFBR</jats:italic>) <jats:italic>1</jats:italic> and <jats:italic>2</jats:italic>. We encountered a patient with LDS, who had severe periodontal tissue destruction indicative of aggressive periodontitis. The patient had a missense mutation in the glycine and serine-rich domain of <jats:italic>TGFBR1</jats:italic> exon 3. This G-to-T mutation at base 563 converted glycine to valine. We established an LDS model knock-in mouse that recapitulated the LDS phenotype. Homozygosity of the mutation caused embryonic lethality and heterozygous knock-in mice showed distorted and ruptured elastic fibers in the aorta at 24 weeks of age and died earlier than wildtype (WT) mice. We stimulated mouse embryonic fibroblasts (MEFs) from the knock-in mouse with TGF-β and examined their responses. The knock-in MEFs showed downregulated <jats:italic>Serpine 1</jats:italic> mRNA expression and phosphorylation of Smad2 to TGF-β compared with WT MEFs. To clarify the influence of TGF-β signaling abnormalities on the pathogenesis or progression of periodontitis, we performed pathomolecular analysis of the knock-in mouse. There were no structural differences in periodontal tissues between WT and LDS model mice at 6 or 24 weeks of age. Micro-computed tomography revealed no significant difference in alveolar bone resorption between WT and knock-in mice at 6 or 24 weeks of age. However, TGF-β-related gene expression was increased significantly in periodontal tissues of the knock-in mouse compared with WT mice. Next, we assessed a mouse periodontitis model in which periodontal bone loss was induced by oral inoculation with the bacterial strain <jats:italic>Porphyromonas gingivalis</jats:italic> W83. After inoculation, we collected alveolar bone and carried out morphometric analysis. <jats:italic>P. gingivalis</jats:italic>-induced alveolar bone loss was significantly greater in LDS model mice than in WT mice. Peritoneal macrophages isolated from <jats:italic>Tgfbr1<jats:sup><jats:italic>G</jats:italic>188V/+</jats:sup></jats:italic> mice showed upregulation of inflammatory cytokine mRNA expression induced by <jats:italic>P. gingivalis</jats:italic> lipopolysaccharide compared with WT macrophages. In this study, we established an LDS mouse model and demonstrated that LDS model mice had elevated susceptibility to <jats:italic>P. gingivalis</jats:italic>-induced periodontitis, probably through TGF-β signal dysfunction. This suggests that TGF-β signaling abnormalities accelerate the pathogenesis or progression of periodontitis.</jats:p>

DOI： 10.3389/fphys.2021.715687

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Accumulating evidence suggests that specific non-coding RNAs exist in many types of malignant tissues, and are involved in cancer invasion and metastasis. However, little is known about the precise roles of non-coding RNAs in squamous cell carcinoma (SQCC) invasion and migration. Recently, the dentin matrix protein-1 (DMP-1) gene locus was identified as a transcriptionally active site in squamous cell carcinoma (SQCC) tissue and cells. However, it is unclear whether RNA associated with cell migration exist at the DMP-1 gene locus in SQCC cells. We identified a novel promoter-associated non-coding RNA in the antisense strand of DMP-1 gene locus, promoter-associated non-coding RNA (panRNA)-DMP-1, by the RACE method in SQCC cells and tissues, and characterized the functions of panRNA-DMP-1 in EGF-driven SQCC cell migration. The inhibition of endogenous panRNA-DMP-1 expression by specific siRNAs and exogenous over-expression of panRNA-DMP-1 resulted in increased and suppressed cellular migration toward EGF in SQCC cells, respectively, and nuclear expression of panRNA-DMP-1 was induced by EGF stimulation. Mechanistically, suppression of panRNA-DMP-1 expression increased EGFR nuclear localization upon EGF treatment and nuclear panRNA-DMP-1 physically interacted with EGFR, which was confirmed by RNA immunoprecipitation assay using a bacteriophage-delivered PP7 RNA labeling system. Furthermore, co-immunoprecipitation assay revealed that suppression of panRNA-DMP-1 stabilized EGFR interaction with STAT3, a known co-transcription factors of EGFR, to induce migratory properties in many cancer cells. Based on these findings, panRNA-DMP-1 is an EGFR-associating RNA that inhibits the EGF-induced migratory properties of SQCC possibly by regulating EGFR nuclear localization and EGFR binding to STAT3.

DOI： 10.1007/s11010-020-04046-5

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Other Link： http://link.springer.com/article/10.1007/s11010-020-04046-5/fulltext.html

Language：English   Publishing type：Research paper (scientific journal)   Publisher：Springer Science and Business Media LLC  

<title>Abstract</title>Apical periodontitis (AP) is an acute or chronic inflammatory disease caused by complex interactions between infected root canal and host immune system. It results in the induction of inflammatory mediators such as chemokines and cytokines leading to periapical tissue destruction. To understand the molecular pathogenesis of AP, we have investigated inflammatory-related genes that regulate AP development. We found here that macrophage-derived CXCL9, which acts through CXCR3, is recruited by progressed AP. The inhibition of CXCL9 by a CXCR3 antagonist reduced the lesion size in a mouse AP model with decreasing IL-1β, IL-6 and TNFα expression. The treatment of peritoneal macrophages with CXCL9 and LPS induced the transmigration and upregulation of osteoclastogenic cytokines such as IL-1β, IL-6 and matrix metalloprotease 2, a marker of activated macrophages. This suggests that the CXCL9-CXCR3 axis plays a crucial role in the development of AP, mediated by the migration and activation of macrophages for periapical tissue destruction. Our data thus show that CXCL9 regulates the functions of macrophages which contribute to AP pathogenesis, and that blocking CXCL9 suppresses AP progression. Knowledge of the principal factors involved in the progression of AP, and the identification of related inflammatory markers, may help to establish new therapeutic strategies.

DOI： 10.1038/s41598-021-82167-7

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Other Link： http://www.nature.com/articles/s41598-021-82167-7

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The spread of root canal infection to surrounding periodontal tissue through accessory root canals reduces the success rate of endodontic treatment. In this case, cone-beam computed tomography revealed a lesion (4 mm from the apex) resulting from an accessory root canal of the maxillary left central incisor. First, non-surgical endodontic treatment was conducted but the sinus tract remained. Surgical preparation of the root cavity was then conducted to remove potentially infected dentin surrounding the accessory root canal. The cavity was filled and the foramen was sealed with resin containing bioactive surface pre-reacted glass (S-PRG) filler. The photopolymerized resin was then contoured and polished. In combination with subsequent supportive non-surgical endodontic treatment, a good clinical outcome with the disappearance of the sinus tract and clinical symptoms such as discomfort and pressure pain and the regeneration of the alveolar bone hanging over the cavity was obtained. In this case, the good clinical outcome may have been due to the dentin-adhesive property and durability of the pre-adhesive system and composite resin. The better biocompatibility of S-PRG fillers presumably facilitated periodontal tissue healing.

DOI： 10.3390/dj8040131

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Pro-inflammatory cytokines prevent bone regeneration in vivo and activation of nuclear factor-κB (NF-κB) signaling has been proposed to lead to suppression of bone morphogenetic protein (BMP)-induced osteogenesis via direct binding of p65 to Smad4 in vitro. Application of a small nuclear acidic protein (MTI-II) and its delivered peptide, MPAID (MTI-II peptide anti-inflammatory drug) has been described to elicit therapeutic potential via strong anti-inflammatory action following the physical association of MTI-II and MPAID with p65. However, it is unclear whether MTI-II attenuates tumor necrosis factor (TNF)-α inhibition of BMP-induced osteogenesis. Herein, we found that TNF-α-mediated suppression of responses associated with BMP4-induced osteogenesis, including expression of the osteocalcin encoding gene Ocn, Smad binding element (SBE)-dependent luciferase activity, alkaline phosphatase activity, and alizarin red S staining were largely restored by MTI-II and MPAID in MC3T3-E1 cells. Mechanistically, MTI-II and MPAID did not inhibit nuclear translocation of p65 or disassociate Smad4 from p65. Further, results from chromatin immunoprecipitation (ChIP) analyses revealed that Smad4 enrichment in cells over-expressing MTI-II and treated with TNF-α was equivalent to that in cells without TNF-α treatment. Alternatively, Smad4 enrichment was considerably decreased following TNF-α treatment in control cells. Moreover, p65 enrichment in the Id-1 promoter SBE was detected only when cells over-expressing MTI-II were stimulated with TNF-α. Overall, our study concludes that MTI-II restored TNF-α-inhibited suppression of BMP-Smad-induced osteogenic differentiation by enhancing accessibility of the Smad4-p65 complex to the SBE rather than by liberating Smad4 from p65.

DOI： 10.1007/s11010-020-03734-6

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DOI： 10.1016/j.archoralbio.2019.104634

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It is well known that dental pulp tissue can evoke some of the most severe acute inflammation observed in the human body. We found that dental pulp cells secrete a factor that induces tumor necrosis factor-α production from macrophages, and designated this factor, dental pulp cell-derived powerful inducer of TNF-α (DPIT). DPIT was induced in dental pulp cells and transported to recipient cells via microvesicles. Treatment of dental pulp cells with a PKR inhibitor markedly suppressed DPIT activity, and weak interferon signals were constitutively activated inside the cells. In recipient macrophages, stimulation with DPIT-containing supernatants from pulp cells resulted in activation of both nuclear factor-κB and MAP kinases like JNK and p38. Proteomics analyses revealed that many stress granule-related proteins were present in supernatants from dental pulp cells as well as microvesicle marker proteins like GAPDH, β-actin, HSPA8, HSPB1, HSPE1, and HSPD1. Furthermore, giant molecule AHNAK and PKR were detected in microvesicles derived from dental pulp cells, and gene silencing of AHNAK in dental pulp cells led to reduced DPIT activity. Thus, it appeared that the core protein of DPIT was PKR, and that PKR was maintained in an active state in stress granule aggregates with AHNAK and transported via microvesicles. The activity of DPIT for TNF-α induction was far superior to that of gram-negative bacterial endotoxin. Therefore, we, report for the first time, that active PKR is transported via microvesicles as stress granule aggregates and induces powerful inflammatory signals in macrophages.

DOI： 10.1038/s41598-019-40046-2

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DOI： 10.1111/iej.13130

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In the oral mechanical environment, periodontal ligament cells (PDL cells) contribute to maintaining periodontal tissue homeostasis. Recent studies showed that exosomes, which are small vesicles secreted by various types of cells, play a pivotal role in cell-to-cell communication in biological processes. We examined the secretion of exosomes from PDL cells stimulated with cyclic stretch and their role in the inflammatory response of macrophages using the human macrophage cell line THP-1 and human primary monocytes/macrophages. We prepared supernatants from human PDL cells (PDL-sup) stimulated with cyclic stretch. The treatment of macrophages with PDL-sup, but not PDL-sup from unstimulated PDL cells, inhibited the production of IL-1β in LPS/nigericin-stimulated macrophages. The pretreatment of PDL cells with GW4869, an inhibitor of exosome secretion, or siRNA for Rab27B, which controls exosome secretion, abrogated the inhibitory effects of PDL-sup. A transmission electron microscopy analysis demonstrated the existence of exosomes with diameters ranging between 30 and 100 nm in PDL-sup, suggesting that exosomes in PDL-sup contribute to this inhibition. An immunofluorescence microscopy analysis revealed that exosomes labeled with PKH67, a fluorescent dye, were incorporated by macrophages as early as 2 h after the addition of exosomes. Purified exosomes inhibited IL-1β production in LPS/nigericin-stimulated macrophages and the nuclear translocation of NF-κB as well as NF-κB p65 DNA-binding activity in LPS-stimulated macrophages, suggesting that exosomes suppress IL-1β production by inhibiting the NF-κB signaling pathway. Our results indicate that PDL cells in mechanical environments contribute to the maintenance of periodontal immune/inflammatory homeostasis by releasing exosomes.

DOI： 10.3389/fimmu.2019.01310

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Authorship：Corresponding author   Language：Japanese   Publishing type：Research paper (scientific journal)   Publisher：(NPO)日本歯周病学会  

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DOI： 10.1016/j.bbrc.2017.12.136

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DOI： 10.1016/j.numecd.2016.11.008

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Authorship：Lead author,　Corresponding author   Language：English   Publisher：Japanese Association for Oral Biology  

DOI： 10.1016/j.job.2016.08.002

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Elsevier BV  

DOI： 10.1016/j.bbrep.2016.04.004

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DOI： 10.1016/j.archoralbio.2015.05.013

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DOI： 10.1002/jbmr.2444

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DOI： 10.1371/journal.pone.0112490

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DOI： 10.1093/ndt/gft449

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Authorship：Lead author,　Corresponding author   Language：Japanese   Publishing type：Research paper (scientific journal)   Publisher：The Japanese Society of Conservative Dentistry  

Purpose: Matrix trioxide aggregate (MTA) has been used for direct pulp-capping as well as perforation repair due to its potent ability to induce hard tissue regeneration. Pulp exposure, usually caused by tooth fracture and carious tooth treatment, involves the physical damage of pulp extracellular matrix. Therefore, when cells invade the damaged space from surrounding tissue, the existence of an adhesive substrate to aid cell proliferation and differentiation of invading cells is the key factor influencing the prognosis of pulp-capping treatment. In this study, we examined the effects of MTA on the adhesion, proliferation and apoptosis of human dental pulp cells (hDPCs) by comparison with composite resin (CR) and glass ionomer cement (GIC).<br> Materials and methods: Ninety-six-well plates were coated with MTA, CR, GIC and fibronectin. Cells were suspended in serum-free medium and then seeded onto coated wells. After 1.5-hr incubation at 37°C, the wells were rinsed to remove non-adherent cells. The number of adherent cells was quantified using the Cell Titer Glo Luminescent Cell Viability Assay Kit. For proliferation assay, these cells were seeded onto MTA, CR and GIC and attached cells were quantified. After 1.5-hr incubation, serum-free medium was replaced with 10% serum medium and cells were cultured onto these substrates for 72 hrs. The ability to induce apoptosis was evaluated by caspase 3/7 activity.<br> Result: MTA facilitated the adhesion of hDPCs, while CR and GIC did not. However, the adhesive effect of MTA was much weaker than that of fibronectin. hDPCs could attach onto MTA but not CR or GIC, even 72 hrs after seeding, although the number of attached cells gradually decreased during the culture period. These observations suggest that MTA could be used as a substrate for the adhesion, but not a scaffold for the proliferation, of hDPCs. A drastic increase of caspase 3/7 activity was observed when hDPCs were cultured onto CR and GIC. In contrast, MTA did not induce it.<br> Conclusions: Compared with CR and GIC, MTA is a good substrate for hDPC adhesion. These differential adhesive effects may be explained by the differential abilities for inducing pro-apoptotic signals in hDPCs. However, the effect of MTA as a cell adhesion substrate was much weaker than that of fibronectin. More importantly, MTA was unable to induce the proliferation of hDPCs on it. Thus, the co-addition of MTA with other adhesive and migratory proteins such as fibronectin may induce better wound healing and pulp tissue regeneration in vivo.

DOI： 10.11471/shikahozon.57.547

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DOI： 10.2174/978160805465711202010221

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DOI： 10.1016/j.archoralbio.2012.03.005

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DOI： 10.1186/1617-9625-10-3

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<jats:p>Tooth and bone specimen require extensive demineralization for careful analysis of cell morphology, as well as gene and protein expression levels. The LacZ gene, which encodes the ß-galactosidase enzyme, is often used as a reporter gene to study gene-structure function, tissue-specific expression by a promoter, cell lineage and fate. This reporter gene is particularly useful for analyzing the spatial and temporal gene expression pattern, by expressing the LacZ gene under the control of a promoter of interest. To analyze LacZ activity, and the expression of other genes and their protein products in teeth and bones, it is necessary to carry out a complete demineralization of the specimen before cutting sections. However, strong acids, such as formic acid used for tooth demineralization, destroy the activities of enzymes including those of ß-galactosidase. Therefore, most protocols currently use mild acids such as 0.1 M ethylene diamine tetra-acetic acid (EDTA) for demineralization of tooth and bone specimen, which require a longer period of treatment for complete demineralization. A method by which hard tissue specimens such as teeth and bones can be rapidly, but gently, decalcified is necessary to save time and effort. Here, we report a suitable method for rapid demineralization of mouse teeth in 0.1M EDTA at 42˚C without any loss of ß-galactosidase activity.</jats:p>

DOI： 10.2174/1874210601004010223

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DOI： 10.1016/j.bbrc.2010.06.112

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DOI： 10.11471/shikahozon.53.73

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DOI： 10.1016/j.matbio.2009.03.006

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DOI： 10.1016/j.matbio.2009.01.005

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DOI： 10.1177/0022034509334749

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Language：Japanese   Publisher：(NPO)日本歯周病学会  

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Language：English   Publisher：(NPO)日本歯周病学会  

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J-GLOBAL

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Language：Japanese  

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Language：English   Publishing type：Research paper, summary (international conference)  

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Language：English   Publishing type：Research paper, summary (international conference)  

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Publishing type：Research paper, summary (national, other academic conference)  

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Language：English   Publisher：(NPO)日本歯周病学会  

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J-GLOBAL

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J-GLOBAL

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Language：Japanese   Publisher：(NPO)日本歯科保存学会  

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Language：Japanese   Publisher：(NPO)日本歯科保存学会  

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Language：Japanese   Publisher：(NPO)日本歯周病学会  

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Language：Japanese   Publisher：(NPO)日本歯科保存学会  

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Language：Japanese   Publisher：(NPO)日本歯科保存学会  

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J-GLOBAL

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J-GLOBAL

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Publishing type：Research paper, summary (national, other academic conference)  

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Language：Japanese   Publisher：(NPO)日本歯周病学会  

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J-GLOBAL

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Publishing type：Research paper, summary (national, other academic conference)  

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Language：Japanese   Publishing type：Research paper, summary (national, other academic conference)   Publisher：The Japanese Society of Conservative Dentistry  

Purpose: Matrix trioxide aggregate (MTA) has been used for direct pulp-capping as well as perforation repair due to its potent ability to induce hard tissue regeneration. Pulp exposure, usually caused by tooth fracture and carious tooth treatment, involves the physical damage of pulp extracellular matrix. Therefore, when cells invade the damaged space from surrounding tissue, the existence of an adhesive substrate to aid cell proliferation and differentiation of invading cells is the key factor influencing the prognosis of pulp-capping treatment. In this study, we examined the effects of MTA on the adhesion, proliferation and apoptosis of human dental pulp cells (hDPCs) by comparison with composite resin (CR) and glass ionomer cement (GIC).<br> Materials and methods: Ninety-six-well plates were coated with MTA, CR, GIC and fibronectin. Cells were suspended in serum-free medium and then seeded onto coated wells. After 1.5-hr incubation at 37°C, the wells were rinsed to remove non-adherent cells. The number of adherent cells was quantified using the Cell Titer Glo Luminescent Cell Viability Assay Kit. For proliferation assay, these cells were seeded onto MTA, CR and GIC and attached cells were quantified. After 1.5-hr incubation, serum-free medium was replaced with 10% serum medium and cells were cultured onto these substrates for 72 hrs. The ability to induce apoptosis was evaluated by caspase 3/7 activity.<br> Result: MTA facilitated the adhesion of hDPCs, while CR and GIC did not. However, the adhesive effect of MTA was much weaker than that of fibronectin. hDPCs could attach onto MTA but not CR or GIC, even 72 hrs after seeding, although the number of attached cells gradually decreased during the culture period. These observations suggest that MTA could be used as a substrate for the adhesion, but not a scaffold for the proliferation, of hDPCs. A drastic increase of caspase 3/7 activity was observed when hDPCs were cultured onto CR and GIC. In contrast, MTA did not induce it.<br> Conclusions: Compared with CR and GIC, MTA is a good substrate for hDPC adhesion. These differential adhesive effects may be explained by the differential abilities for inducing pro-apoptotic signals in hDPCs. However, the effect of MTA as a cell adhesion substrate was much weaker than that of fibronectin. More importantly, MTA was unable to induce the proliferation of hDPCs on it. Thus, the co-addition of MTA with other adhesive and migratory proteins such as fibronectin may induce better wound healing and pulp tissue regeneration in vivo.

DOI： 10.11471/shikahozon.57.547

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Other Link： http://search.jamas.or.jp/link/ui/2015182150

Publishing type：Research paper, summary (national, other academic conference)  

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J-GLOBAL

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Language：Japanese   Publisher：(NPO)日本歯周病学会  

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Language：Japanese   Publisher：(NPO)日本歯周病学会  

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Publishing type：Research paper, summary (national, other academic conference)  

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Publishing type：Research paper, summary (national, other academic conference)  

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Language：Japanese   Publishing type：Research paper, summary (national, other academic conference)   Publisher：(NPO)日本歯周病学会  

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Publishing type：Research paper, summary (national, other academic conference)  

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Language：Japanese   Publisher：(NPO)日本歯周病学会  

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Language：Japanese   Publishing type：Research paper, summary (international conference)   Publisher：JAPANESE SOCIETY OF PERIODONTOLOGY  

DOI： 10.14833/amjsp.2011s.0.23.0

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Publishing type：Research paper, summary (international conference)  

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Publishing type：Research paper, summary (international conference)  

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Publishing type：Research paper, summary (international conference)  

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Language：Japanese  

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Language：Japanese   Publisher：日本炎症・再生医学会  

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Language：Japanese  

DOI： 10.14833/amjsp.2006s.0.71.0

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Language：Japanese   Publisher：(NPO)日本歯周病学会  

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Language：Japanese   Publisher：日本炎症・再生医学会  

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Language：Japanese   Publisher：(NPO)日本歯周病学会  

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Publishing type：Research paper, summary (international conference)  

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Publishing type：Research paper, summary (international conference)  

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Language：Japanese   Publisher：(NPO)日本歯周病学会  

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Language：Japanese   Publisher：(NPO)日本歯周病学会  

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CiNii Books

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Language：Japanese   Publisher：(NPO)日本歯周病学会  

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CiNii Books

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Language：Japanese   Publisher：(NPO)日本歯周病学会  

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Language：Japanese   Publisher：(NPO)日本歯周病学会  

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Language：Japanese   Publisher：大阪大学歯学会  

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Language：English   Presentation type：Oral presentation (invited, special)  

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Language：English   Presentation type：Oral presentation (invited, special)  

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Language：Japanese   Presentation type：Oral presentation (invited, special)  

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Language：Japanese   Presentation type：Oral presentation (invited, special)  

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Language：English   Presentation type：Oral presentation (invited, special)  

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