研究者詳細 - 山本　直史

論文 - 山本　直史

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Modulation of microenvironment for controlling the fate of periodontal ligament cells: the role of Rho/ROCK signaling and cytoskeletal dynamics 招待査読

Tadashi Yamamoto, Yuki Ugawa, Mari Kawamura, Keisuke Yamashiro, Shinsuke Kochi, Hidetaka Ideguchi, Shogo Takashiba

Journal of Cell Communication and Signaling 12 ( 1 ) 369 - 378 2018年3月

担当区分：筆頭著者記述言語：英語出版者・発行元：Springer Netherlands

© 2017, The International CCN Society. Cells behave in a variety of ways when they perceive changes in their microenvironment; the behavior of cells is guided by their coordinated interactions with growth factors, niche cells, and extracellular matrix (ECM). Modulation of the microenvironment affects the cell morphology and multiple gene expressions. Rho/Rho-associated coiled-coil-containing protein kinase (ROCK) signaling is one of the key regulators of cytoskeletal dynamics and actively and/or passively determines the cell fate, such as proliferation, migration, differentiation, and apoptosis, by reciprocal communication with the microenvironment. During periodontal wound healing, it is important to recruit the residential stem cells into the defect site for regeneration and homeostasis of the periodontal tissue. Periodontal ligament (PDL) cells contain a heterogeneous fibroblast population, including mesenchymal stem cells, and contribute to the reconstruction of tooth-supporting tissues. Therefore, bio-regeneration of PDL cells has been the ultimate goal of periodontal therapy for decades. Recent stem cell researches have shed light on intrinsic ECM properties, providing paradigm shifts in cell fate determination. This review focuses on the role of ROCK activity and the effects of Y-27632, a specific inhibitor of ROCK, in the modulation of ECM-microenvironment. Further, it presents the current understanding of how Rho/ROCK signaling affects the fate determination of stem cells, especially PDL cells. In addition, we have also discussed in detail the underlying mechanisms behind the reciprocal response to the microenvironment.

DOI： 10.1007/s12079-017-0425-3

感染、炎症、機能の観点から見た口腔検査指標の歯周病治療に対する有用性パイロット研究査読

畑中加珠, 片山広大, 井上裕貴, 坂井田京佑, 清水由梨香, 鈴木里紗, 高木美奈, 山本直史, 高柴正悟

日本口腔検査学会雑誌 10 ( 1 ) 58 - 63 2018年3月

記述言語：日本語出版者・発行元：(一社)日本口腔検査学会

目的:歯周病治療の効果を「感染」、「炎症」、そして「機能」の3つの観点から評価して、治療効果を示す指標を探った。方法:歯周病治療を終えて安定期治療に至った7症例において、歯周局所の歯周病原細菌数と9菌種の血清IgG抗体価、歯周ポケット深さとプロービング時の出血から算出する歯周炎症表面積(PISA)、そして咀嚼機能の簡易指標として残存歯の動揺度を測定した。これら4つの指標の変動を治療時期の一定ポイントで調べ、それらの相互関係を検討した。結果:歯周治療が進むにしたがって、PISA、歯の動揺度およびPorphyromonas gingivalis(Pg)に対する抗体価の数値は低下し、7症例の各時期における平均値をとると、経時的な有意差が認められた(P<0.05)。しかしながら、他の菌種に対する抗体価や歯周局所の細菌数には増加するものもみられた。結論:「感染」、「炎症」、「機能」の3つの観点から歯周病治療の効果を評価する指標として、抗Pg抗体価、PISA、そして歯の動揺度を用いることは有用であった。(著者抄録)

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Effects of Lectins on initial attachment of cariogenic Streptococcus mutans 査読

Takashi Ito, Yasuhiro Yoshida, Yasuyoshi Shiota, Yuki Ito, Tadashi Yamamoto, Shogo Takashiba

Glycoconjugate Journal 35 ( 1 ) 41 - 51 2018年2月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Springer New York LLC

© 2017, Springer Science+Business Media, LLC. Oral bacteria initiate biofilm formation by attaching to tooth surfaces via an interaction of a lectin-like bacterial protein with carbohydrate chains on the pellicle. This study aimed to find naturally derived lectins that inhibit the initial attachment of a cariogenic bacterial species, Streptococcus mutans (S. mutans), to carbohydrate chains in saliva in vitro. Seventy kinds of lectins were screened for candidate motifs that inhibit the attachment of S. mutans ATCC 25175 to a saliva-coated culture plate. The inhibitory effect of the lectins on attachment of the S. mutans to the plates was quantified by crystal violet staining, and the biofilm was observed under a scanning electron microscope (SEM). Surface plasmon resonance (SPR) analysis was performed to examine the binding of S. mutans to carbohydrate chains and the binding of candidate lectins to carbohydrate chains, respectively. Moreover, binding assay between the biotinylated-lectins and the saliva components was conducted to measure the lectin binding. Lectins recognizing a salivary carbohydrate chain, Galβ1-3GalNAc, inhibited the binding of S. mutans to the plate. In particular, Agaricus bisporus agglutinin (ABA) markedly inhibited the binding. This inhibition was confirmed by SEM observation. SPR analysis indicated that S. mutans strongly binds to Galβ1-3GalNAc, and ABA binds to Galβ1-3GalNAc. Finally, the biotinylated Galβ1-3GalNAc-binding lectins including ABA demonstrated marked binding to the saliva components. These results suggest that ABA lectin inhibited the attachment of S. mutans to Galβ1-3GalNAc in saliva and ABA can be useful as a potent inhibitor for initial attachment of oral bacteria and biofilm formation.

DOI： 10.1007/s10719-017-9795-2

IS1598 (IsPg4) distributed to abscess-forming strains of Porphyromonas gingivalis may enhance virulence through upregulation of nrdD-like gene expression 査読

Norihiro Sonoi, Hiroshi Maeda, Toshimitsu Murauchi, Tadashi Yamamoto, Kazuhiro Omori, Susumu Kokeguchi, Koji Naruishi, Shogo Takashiba

New Microbiologica 41 ( 1 ) 52 - 60 2018年1月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Luigi Ponzio e figlio Editori

©2018 by EDIMES-Edizioni Internazionali Srl. All rights reserved. An insertion sequence, IS1598 (IsPg4) has been found in virulent strains of Porphyromonas gingivalis in a murine abscess model. The present study was performed to investigate the effects of genetic rearrangements by IS1598 on the phenotypic characteristics of the virulent strains. For this purpose, we searched for a common insertion site of IS1598 among the virulent strains. Through cloning and database search, a common insertion site was identified beside an nrdD-like gene in the virulent FDC 381, W83 and W50 strains. In this region, predicted promoters of the nrdD-like gene and IS1598 are located in tandem, and accumulation of nrdD-like gene mRNA was 5-fold higher in virulent strains (W83, W50, FDC 381) than avirulent strains (ATCC33277, SU63, SUNY1021, ESO59 without IS1598). The role of the nrdD-like gene in virulence of P. gingivalis was investigated by constructing a nrdD-deficient mutant. In the murine abscess model, the parental W83 strain produced necrotic abscesses, while the nrdD-deficient mutant had almost lost this ability. Insertion of IS1598 into the nrdD-like gene promoter region may be related to the phenotypic differences in virulence among P. gingivalis strains through upregulation of the expression of this gene.

Expression of optineurin isolated from rat-injured dental pulp and the effects on inflammatory signals in normal rat kidney cells

Tadashi Yamamoto

Odontology 106 ( 2 ) 2018年

Aggregatibacter actinomycetemcomitans regulates the expression of integrins and reduces cell adhesion via integrin α5 in human gingival epithelial cells. 査読

Kochi S, Yamashiro K, Hongo S, Yamamoto T, Ugawa Y, Shimoe M, Kawamura M, Hirata-Yoshihara C, Ideguchi H, Maeda H, Takashiba S

Molecular and cellular biochemistry 436 ( 1-2 ) 39 - 48 2017年12月

Assessment of pathogenesis of infective endocarditis by plasma IgG antibody titer test against periodontal bacteria. 査読国際誌

Isoshima D, Yamashiro K, Matsunaga K, Shinobe M, Nakanishi N, Nakanishi I, Omori K, Yamamoto T, Takashiba S

Clinical case reports 5 ( 10 ) 1580 - 1586 2017年10月

Rho-kinase regulates extracellular matrix-mediated osteogenic differentiation of periodontal ligament cells 査読

Yuki Ugawa, Tadashi Yamamoto, Mari Kawamura, Keisuke Yamashiro, Masayuki Shimoe, Kazuya Tomikawa, Shoichi Hongo, Hiroshi Maeda, Shogo Takashiba

CELL BIOLOGY INTERNATIONAL 41 ( 6 ) 651 - 658 2017年6月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：WILEY

The periodontal ligament (PDL) cells contain heterogeneous mesenchymal cell populations, which have the ability to differentiate into cells that produce adjacent mineralized tissues and abundant extracellular matrix (ECM). ECM is essential not only for the homeostasis of the periodontal tissue, but also for controlling the differentiation of the PDL cells. The process of differentiation involves mechanotransduction, which links the ECM to the cytoskeleton. The present study investigated the roles of Rho-associated coiled-coil containing protein kinase (ROCK) signaling, a crucial regulator of the cytoskeleton, during ECM-mediated osteogenic differentiation of PDL cells in vitro. The PDL cells were isolated from human periodontal ligaments of extracted teeth and cultured in osteogenic medium with or without Y-27632, a pharmacological inhibitor of ROCK. ECMcoated plates were used for ECM-mediated differentiation. The osteogenic phenotype was evaluated at different time points by real-time RT-PCR for the gene encoding alkaline phosphatase (ALP) and an ALP activity assay. The effects of ROCK on cytoskeletal changes and ECM synthesis were examined by immunofluorescence analysis. Y-27632 significantly inhibited ALP at the mRNA and protein activity levels in the late stage of differentiation; concomitantly, the actin filament content and the extracellular levels of collagen-I and fibronectin were markedly decreased by Y-27632. Exogenous collagen-I and fibronectin temporally increased ALP activity, with fibronectin showing a more pronounced effect. Importantly, ECM-mediated differentiation was almost completely inhibited by Y-27632. These findings indicated that ECM-mediated differentiation is dependent on ROCK signaling, and ROCK signaling contributes to the establishment of the ECM microenvironment for PDL cell differentiation.

DOI： 10.1002/cbin.10769

Smad2 overexpression enhances adhesion of gingival epithelial cells 査読

Shoichi Hongo, Tadashi Yamamoto, Keisuke Yamashiro, Masayuki Shimoe, Kazuya Tomikawa, Yuki Ugawa, Shinsuke Kochi, Hidetaka Ideguchi, Hiroshi Maeda, Shogo Takashiba

ARCHIVES OF ORAL BIOLOGY 71 46 - 53 2016年11月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：PERGAMON-ELSEVIER SCIENCE LTD

Objective: Gingival epithelial cells play an important role in preventing the initiation of periodontitis, by their hemidesmosomal adhesion to the tooth root surface. Adhesion requires integrin-extracellular matrix (ECM) interactions that are intricately regulated by transforming growth factor-beta (TGF-beta) signaling. However, the mechanisms underlying the interplay between adhesion molecules and TGF-beta, especially the respective roles of Smad2 and Smad3, remain elusive. In this study, we examined the effects of Smad overexpression on gingival epithelial cell adhesion and expression profiles of integrin and ECM-related genes.
Methods: Human gingival epithelial cells immortalized by the SV40 T-antigen were transfected with Smad2- and Smad3-overexpression vectors. A cell adhesion assay involving fluorescence detection of attached cells was performed using the ArrayScan imaging system. Real-time PCR was performed to examine the kinetics of integrin and ECM gene expression. In vitro and in vivo localization of adhesion molecules was examined by immunofluorescence analysis.
Results: By using SB431542, a specific inhibitor of the TGF-beta type I receptor, Smad2/3 signaling was confirmed to be dominant in TGF-beta 1-induced cell adhesion. The Smad2-transfectant demonstrated higher potency for cell adhesion and integrin expression (alpha 2, alpha 5, beta 4, and beta 6) than the Smad3-transfectant, whereas little or no change in ECM expression was observed in either transfectant. Moreover, the gingival epithelium of transgenic mice that overexpressed Smad2 driven by the keratin 14 promoter showed increased integrin alpha 2 expression.
Conclusion: These findings indicate the crucial role of Smad2 in increased adhesion of gingival epithelial cells via upregulation of integrin alpha 2. (C) 2016 Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.archoralbio.2016.06.025

Osteogenic differentiation regulated by Rho-kinase in periodontal ligament cells 査読

Tadashi Yamamoto, Yuki Ugawa, Keisuke Yamashiro, Masayuki Shimoe, Kazuya Tomikawa, Shoichi Hongo, Shinsuke Kochi, Hidetaka Ideguchi, Hiroshi Maeda, Shogo Takashiba

DIFFERENTIATION 88 ( 2-3 ) 33 - 41 2014年9月

担当区分：筆頭著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：ELSEVIER SCI LTD

The periodontal ligament is a multifunctional soft connective tissue, which functions not only as a cushion supporting the teeth against occlusal force, but is also a source of osteogenic cells that can regenerate neighboring hard tissues. Periodontal ligament cells (PDL cells) contain heterogeneous cell populations, including osteogenic cell progenitors. However, the precise mechanism underlying the differentiation process remains elusive. Cell differentiation is regulated by the local biochemical and mechanical microenvironment that can modulate gene expression and cell morphology by altering actin cytoskeletal organization mediated by Rho-associated, coiled-coil containing protein kinase (ROCK). To determine its role in PDL cell differentiation, we examined the effects of ROCK on cytoskeletal changes and kinetics of gene expression during osteogenic differentiation. PDL cells were isolated from human periodontal ligament on extracted teeth and cultured in osteogenic medium for 14 days. Y-27632 was used for ROCK inhibition assay. Osteogenic phenotype was determined by monitoring alkaline phosphatase (ALP) activity and calcium deposition by Alizarin Red staining. ROCK-induced cytoskeletal changes were examined by immunofluorescence analysis of F-actin and myosin light chain 2 (MLC2) expression. Real-time PCR was performed to examine the kinetics of osteogenic gene expression. F-actin and phospho-MLC2 were markedly induced during osteogenic differentiation, which coincided with upregulation of ALP activity and mineralization. Subsequent inhibition assay indicated that Y-27632 significantly inhibited F-actin and phospho-MLC2 expression in a dose dependent manner with concomitant partial reversal of the PDL cell osteogenic phenotype. PCR array analysis of osteogenic gene expression indicated that extracellular matrix genes, such as fibronectin 1, collagen type I and Ill, and biglycan, were significantly downregulated by Y27632. These findings indicated crucial effects of ROCK in cytoskeletal reorganization and differentiation of PDL cells toward osteogenic cells. ROCK contributes to induction of osteogenic differentiation by synergistic increases in extracellular matrix gene expression in PDL cells. (C) 2014 International Society of Differentiation, Published by Elsevier B.V. All rights reserved.

DOI： 10.1016/j.diff.2014.09.002

Overexpression of Smad2 inhibits proliferation of gingival epithelial cells 査読

M. Shimoe, T. Yamamoto, N. Shiomi, K. Tomikawa, S. Hongo, K. Yamashiro, T. Yamaguchi, H. Maeda, S. Takashiba

JOURNAL OF PERIODONTAL RESEARCH 49 ( 3 ) 290 - 298 2014年6月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：WILEY-BLACKWELL

Background and Objective
Spatiotemporal inhibition of apical migration and proliferation of gingival epithelium are significant factors involved in periodontal regeneration. Transforming growth factor beta (TGF-beta) is important in multiple aspects of wound healing, and Smad2, a downstream transcription factor of TGF-beta, has an inhibitory effect on re-epithelialization during gingival wound healing. Therefore, we investigated the effects on migration and proliferation status, and intra/extracellular signaling regulated by Smad2 overexpression in gingival epithelial cells.
Material and Methods
Gingival epithelial cells were isolated from the palatal gingival tissue of transgenic mice overexpressing Smad2 driven by the Keratin14 promoter. Smad2 expression was identified by western blotting and immunofluorescence analysis. Scratch assay and 5-bromo-2 '-deoxyuridine staining were performed to assess cell migration and proliferation. To inactivate TGF-beta type I receptor, the cultures were supplemented with SB431542. Secreted TGF-beta was quantified by ELISA. Smad2 target gene expression was examined by real-time RT-PCR and in vivo immunofluorescence analysis of gingival junctional epithelium.
Results
Smad2-overexpressing cells were confirmed to have significant phosphorylated Smad2 in the nucleus. Scratch assay and 5-bromo-2 '-deoxyuridine staining indicated that Smad2-overexpressing cells showed no significant differences in migration, but had reduced proliferation rates compared to wild-type controls. SB431542 significantly inhibited Smad2 phosphorylation, which coincided with restoration of the proliferation rate in Smad2-overexpressing cells. ELISA of TGF-beta release did not show any differences between genotypes. The cell cycle inhibitors, p15 and p21, showed significant upregulation in Smad2-overexpressing cells compared to wild-type controls. Moreover, junctional epithelium of the transgenic mice showed increased expression of P-Smad2, p15 and p21.
Conclusion
The signaling activation triggered by overexpression of Smad2 was dependent on TGF-beta type I receptor, and the activated Smad2 increased p15 and p21 expression, responsible for inhibiting cell cycle entry, resulting in antiproliferative effects on gingival epithelial cells. Understanding of Smad2-induced signaling would be useful for possible clinical application to regulate gingival epithelial downgrowth.

DOI： 10.1111/jre.12106

細菌感染度を評価しながら包括的歯周治療を行った広汎型侵襲性歯周炎患者の一症例査読

冨川和哉, 河野隆幸, 山本直史, 岩本義博, 下江正幸, 山口知子, 本郷昌一, 宮本学, 前田博史, 高柴正悟

日本歯周病学会会誌 55 ( 4 ) 340 - 348 2013年12月

記述言語：日本語出版者・発行元：(NPO)日本歯周病学会

歯周病に対する感受性が高い侵襲性歯周炎患者の治療においては、徹底した感染のコントロールによる疾患活動性の抑制が必要である。とりわけ、歯列不正を伴う歯周炎患者に矯正治療を行うことは、長期に渡る感染のコントロールを行う上で非常に有効である。しかし、歯周炎患者に矯正治療を行う場合に、感染がコントロールされている歯周状態かどうかを決定する基準は未だ明確でない。今回報告するのは、Agregatibacter actinomycetemcomitans(Aa)の感染が主な病態形成因子と考えられる、初診時22歳の広汎型侵襲性歯周炎患者に対して、矯正治療を含めた包括的歯周治療を行った症例である。初診以降、臨床計測値の変化に加えて、細菌DNA検査と歯周病原細菌に対する血清IgG抗体価検査を用いて、歯周病原細菌の感染度を評価した。その結果、歯周外科治療を含む感染源除去によって、細菌DNA検査でAaが検出されず、Aaに対する血清IgG抗体価が健常者レベルに推移したことを矯正治療移行前に確認した。すなわち、歯周病原細菌感染度が低下したと判断した後に、矯正治療を行うことで、現在まで良好なSPTを維持している。本症例においては、Aaの感染度の低下を細菌DNA検査と血清IgG抗体価を用いて評価することによって、客観的な根拠をもって矯正治療に移行できたと考える。(著者抄録)

DOI： 10.2329/perio.55.340

その他リンク： https://search.jamas.or.jp/index.php?module=Default&action=Link&pub_year=2013&ichushi_jid=J01095&link_issn=&doc_id=20140117230007&doc_link_id=%2Fcp9perlo%2F2013%2F005504%2F008%2F0340-0348%26dl%3D0&url=https%3A%2F%2Fwww.medicalonline.jp%2Fjamas.php%3FGoodsID%3D%2Fcp9perlo%2F2013%2F005504%2F008%2F0340-0348%26dl%3D0&type=MedicalOnline&icon=https%3A%2F%2Fjk04.jamas.or.jp%2Ficon%2F00004_2.gif

Serum antibody response to group II chaperonin from Methanobrevibacter oralis and human chaperonin CCT 査読

Kimito Hirai, Hiroshi Maeda, Kazuhiro Omori, Tadashi Yamamoto, Susumu Kokeguchi, Shogo Takashiba

PATHOGENS AND DISEASE 68 ( 1 ) 12 - 19 2013年6月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：WILEY-BLACKWELL

Both group I (HSP60) and group II (CCT) chaperonins are targets of autoantibodies. Autoimmune reactions to HSP60 have been well characterized, while immune reactions to group II chaperonin have not been clarified. Methanobrevibacter oralis is a suspected periodontal pathogen with group II chaperonin. In this study, serum responses to M.oralis chaperonin, human HSP60, and CCT subunits were examined using sera from patients with periodontitis and autoimmune diseases. In comparison with healthy controls, periodontitis patients showed significantly higher responses to CCT4 and CCT8 on dot blot analysis. Signals for CCT3 and CCT8 in autoimmune disease patients were significantly higher than in controls. Significant differences were also demonstrated by Western blotting in anti-CCT4 response in both patient groups. All subjects showed strong reactivity to M.oralis chaperonin and faint signals to human HSP60. Autoantibodies were raised against CCT rather than HSP60; and CCT3, CCT4, and CCT8 were shown to be the main targets. Host immune systems may be frequently exposed to chaperonins of Archaea in various habitats. Although further studies of the cross-reactivity between M.oralis chaperonin and human CCT are required, anti-CCT autoantibodies may be involved in the pathogenesis of periodontitis and autoimmune diseases.

DOI： 10.1111/2049-632X.12041

Medical microbiological approach to Archaea in oral infectious diseases 査読

Hiroshi Maeda, Kimito Hirai, Junji Mineshiba, Tadashi Yamamoto, Susumu Kokeguchi, Shogo Takashiba

Japanese Dental Science Review 49 ( 2 ) 72 - 78 2013年5月

記述言語：英語

Recent advances in molecular biological techniques have yielded large amounts of information regarding the oral microflora. The microbiological communities were shown to be more diverse than previously thought and to include a number of previously uncharacterized microorganisms. The range of research targets of microorganisms associated with oral diseases has been expanded to include these unknown or uncharacterized organisms. These organisms include the Archaea. A series of recent reports suggested these microorganisms to be potential pathogens involved in periodontitis and apical periodontitis mainly based on the detection frequency or their increased numbers in diseased sites in association with the severity or symptoms of disease. However, it cannot be concluded that Archaea are oral pathogens based on such circumstantial evidence. Further studies are required to investigate the potential pathogenic mechanisms of action of these organisms. This will require investigation of the antigenic properties of the Archaea and synergism with other established oral pathogens. Especially, studies of the host immune response will provide insight into the medical impact of Archaea as suspected pathogens. © 2013 Japanese Association for Dental Science.

DOI： 10.1016/j.jdsr.2013.01.002

Smad2 Decelerates Re-epithelialization during Gingival Wound Healing 査読

K. Tomikawa, T. Yamamoto, N. Shiomi, M. Shimoe, S. Hongo, K. Yamashiro, T. Yamaguchi, H. Maeda, S. Takashiba

JOURNAL OF DENTAL RESEARCH 91 ( 8 ) 764 - 770 2012年8月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：SAGE PUBLICATIONS INC

During periodontal regeneration, inhibition of gingival downgrowth is necessary to promote migration of mesenchymal cells into the defects. Transforming growth factor (TGF)-beta is a pleiotropic cytokine that has numerous cell functions, including regulation of epithelial growth. Recent studies have shown that Smad2, a downstream transcription factor of TGF-beta, plays crucial roles in wound healing in the epithelia. Therefore, we investigated the effects of Smad2 overexpression on re-epithelialization of gingival wounds. Transgenic mice overexpressing smad2 driven by the keratin 14 promoter (k14-smad2) were confirmed to have significant Smad2 phosphorylation in gingival basal epithelia. Punch wounds were made in the palatal gingiva, and wound healing was assessed histologically for 7 days. Re-epithelialization was significantly retarded on day 2, while collagen deposition was enhanced on day 7 in k14-smad2 compared with wild-type mice. Moreover, expression of keratin 16 (K16), an indicator of keratinocyte migration, was significantly inhibited in wound-edge keratinocytes in k14-smad2. The inhibition of K16 coincided with the induction of Smad2 in the corresponding epithelia, while BrdU incorporation was unaffected. These results indicated that Smad2 has inhibitory effects in regulating keratinocyte migration during gingival wound healing. TGF-beta/Smad2 signaling mediating alteration of K16 expression must be tightly regulated during periodontal regeneration.

DOI： 10.1177/0022034512451449

The promotional effect of IL-22 on mineralization activity of periodontal ligament cells 査読

Nahoko Kato-Kogoe, Toshihiro Nishioka, Mutsuki Kawabe, Fusa Kataoka, Koji Yamanegi, Naoko Yamada, Masaki Hata, Tadashi Yamamoto, Keiji Nakasho, Masahiro Urade, Nobuyuki Terada, Hideki Ohyama

CYTOKINE 59 ( 1 ) 41 - 48 2012年7月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD

Objectives: Interleukin (IL)-22 acts on non-immune cells to induce anti-microbial responses, protection from tissue damage, and enhance cell regeneration. However, little is known about the involvement of IL-22 in periodontal biology. This study investigated the biological effects of IL-22 on periodontal ligament (PDL) cells as part of studies to assess the involvement of IL-22 in periodontal disease.
Materials and methods: Gene expression levels of IL-22 and its receptors in PDL cells and gingival tissue samples were evaluated by real-time PCR. Proliferative responses and mineralized-matrix forming activities of PDL cells were examined in the presence and absence of IL-22.
Results: In contrast to the expression of IL-22 receptors detected in PDL tissues and their cell lines, gingival tissues showed modest or no gene expressions of IL-22. The production of several cytokines including IL-11, IL-8 and CCL2 was upregulated by IL-22 treatment of PDL cells in a dose-dependent manner. IL-22 treatment had no effect on the proliferative response in PDL cells. Meanwhile, IL-22 precipitated mineralized nodule formation and induced gene expressions of RUNX2,MSX2 and osteocalcin in PDL cells, suggesting that IL-22 enhances the mineralized matrix-forming activities of PDL cells.
Conclusion: IL-22 has the potential to promote mineralizing activity in PDL cells and to develop appropriate regenerative therapy. (C) 2012 Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.cyto.2012.03.024

血清抗体価検査を応用して治療した広汎型侵襲性歯周炎患者の15年間の経過査読

冨川和哉, 岩本義博, 大江丙午, 新井英雄, 山本直史, 前田博史, 高柴正悟

日本歯周病学会会誌 54 ( 2 ) 193 - 202 2012年6月

記述言語：日本語出版者・発行元：(NPO)日本歯周病学会

歯周病に対する感受性が高い侵襲性歯周炎患者の治療においては，徹底した感染コントロールを行い，疾患活動性を抑制する事が要求される。近年，歯周病原細菌の感染量や歯周炎の活動性を評価するために，PCR 法を応用した細菌 DNA 検査や血清抗体価検査が活用されるようになってきた。今回報告するのは，好中球貪食能が低下傾向であった広汎型侵襲性歯周炎患者の症例である。患者は 25 歳の女性であり，歯周基本治療，歯周組織再生療法を含む歯周外科治療，そして最終補綴治療を経て SPT に移行し，初診から 15 年間にわたって，臨床計測値の変化および歯周病原細菌に対する血清 IgG 抗体価の変動をモニタリングしている。これによって，SPT 期間中に歯周組織の破壊が進行した 26 の臨床所見と血清 IgG 抗体価の変動を捉えることができた。26 以外の歯周組織は SPT 期間を通して安定した状態を示していたが，血清 IgG 抗体価は 26 の組織破壊の進行と連動して高値を示した。本症例においては局所的な歯周炎の活動性が血清 IgG 抗体価検査によって評価できたと考える。日本歯周病学会会誌（日歯病誌）54(2)：193-202, 2012

DOI： 10.2329/perio.54.193

Chronic periodontitis with multiple risk factor syndrome: a case report. 査読国際誌

Masayuki Shimoe, Tadashi Yamamoto, Yoshihiro Iwamoto, Nobuyuki Shiomi, Hiroshi Maeda, Fusanori Nishimura, Shogo Takashiba

Journal of the International Academy of Periodontology 13 ( 2 ) 40 - 7 2011年7月

記述言語：英語掲載種別：研究論文（学術雑誌）

OBJECTIVE: Multiple risk factor syndrome is a clustering of cardiovascular risk factors, such as diabetes, dyslipidemia, hypertension, and obesity associated epidemiologically with insulin resistance. This report describes the clinical course of a patient suffering from severe periodontitis with multiple risk factor syndrome, and discusses the association between periodontal infection and systemic health. METHODS: The patient had a history of type 2 diabetes, dyslipidemia, and hypertension for over 10 years. At baseline, her hemoglobin A1 c was 8.1%. However, she had no diabetic complications except periodontitis. The IgG antibody titers against Porphyromonas gingivalis FDC 381 and SU63 were elevated above the mean of healthy subjects +2 standard deviations. Intensive periodontal treatment, including periodontal surgery, was performed to reduce periodontal infection and bacteremia. Her systemic and periodontal conditions were evaluated longitudinally for 10 years. RESULTS: Following periodontal treatment, antibody titers against Porphyromonas gingivalis and hemoglobin A1c values were significantly improved. The other clinical data and medication for her systemic condition also remained stable during supportive periodontal therapy. However, she developed myocardial infarction, and showed continuous deterioration of hemoglobin A1 c level and periodontitis. CONCLUSION: The long-term clustering of risk factors, such as diabetes, dyslipidemia, hypertension, and periodontitis, are associated with the development of myocardial infarction. Treatment of systemic conditions in combination with comprehensive periodontal treatment is important in management of patients with multiple risk factor syndrome.

Rapid detection of mecA and spa by the loop-mediated isothermal amplification (LAMP) method. 査読国際誌

Y Koide, H Maeda, K Yamabe, K Naruishi, T Yamamoto, S Kokeguchi, S Takashiba

Letters in applied microbiology 50 ( 4 ) 386 - 92 2010年4月

記述言語：英語掲載種別：研究論文（学術雑誌）

AIM: To develop a detection assay for staphylococcal mecA and spa by using loop-mediated isothermal amplification (LAMP) method. METHODS AND RESULTS: Staphylococcus aureus and other related species were subjected to the detection of mecA and spa by both PCR and LAMP methods. The LAMP successfully amplified the genes under isothermal conditions at 64 degrees C within 60 min, and demonstrated identical results with the conventional PCR methods. The detection limits of the LAMP for mecA and spa, by gel electrophoresis, were 10(2) and 10 cells per tube, respectively. The naked-eye inspections were possible with 10(3) and 10 cells for detection of mecA and spa, respectively. The LAMP method was then applied to sputum and dental plaque samples. The LAMP and PCR demonstrated identical results for the plaque samples, although frequency in detection of mecA and spa by the LAMP was relatively lower for the sputum samples when compared to the PCR methods. CONCLUSION: Application of the LAMP enabled a rapid detection assay for mecA and spa. The assay may be applicable to clinical plaque samples. SIGNIFICANCE AND IMPACT OF THE STUDY: The LAMP offers an alternative detection assay for mecA and spa with a great advantage of the rapidity.

DOI： 10.1111/j.1472-765X.2010.02806.x

Highly expressed genes in a rough-colony-forming phenotype of Aggregatibacter actinomycetemcomitans: implication of a mip-like gene for the invasion of host tissue 査読

Takemasa Maeda, Hiroshi Maeda, Kokoro Yamabe, Junji Mineshiba, Ichiro Tanimoto, Tadashi Yamamoto, Koji Naruishi, Susumu Kokeguchi, Shogo Takashiba

FEMS IMMUNOLOGY AND MEDICAL MICROBIOLOGY 58 ( 2 ) 226 - 236 2010年3月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：WILEY-BLACKWELL

Aggregatibacter actinomycetemcomitans, a potent pathogen of periodontitis, typically grows as a rough and adherent colony on primary isolated cultures. The colony transforms into a smooth phenotype during repeated subculture. In this study, we aimed to identify highly expressed genes in the rough-colony-forming phenotype for isolation of host-induced genes. Using a cDNA-subtractive hybridization technique, three genes, homologous to a macrophage infectivity potentiator gene (mip), peroxiredoxin gene (prx) and outer membrane protein gene (ompA), were identified. The expression levels of these genes in the rough-colony-forming phenotype were 4-10-fold higher as compared with the smooth-colony-forming phenotype. Attention was focused on the mip-like gene, and a recombinant protein and a deficient mutant were constructed. The recombinant protein reacted with sera from patients with periodontitis, suggesting the production of the Mip-like protein in periodontal lesions. Viable quantitative invasion assay demonstrated that the viable cell counts of the wild-type strain that invaded HeLa cells were more than fourfold as compared with the mip-deficient mutant. The expression of the mip-like gene, prx-like gene and ompA-like gene may be enhanced in the host, and the mip-like gene may play an important role in the infection of A. actinomycetemcomitans, especially in its invasion of the epithelium.

DOI： 10.1111/j.1574-695X.2009.00624.x

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