研究者詳細 - 窪木　拓男

OstemiR: A Novel Panel of MicroRNA Biomarkers in Osteoblastic and Osteocytic Differentiation from Mesencymal Stem Cells 査読

Eguchi Takanori, Hara Emilio Satoshi, Ono Mitsuaki, Watanabe Ken, Kuboki Takuo, Calderwood Stuart

Faseb Journal 29 2015年

BMP2アンタゴニストを阻害するL51Pは骨形成を促進する国際誌

Bone 69 165 - 73 2014年12月

Comparison of platelet serotonin transporter activity in subjects with severe sleep bruxism and control.

Hajime Minakuchi, Chiharu Sogawa, Emilio Satoshi Hara, Haruna Miki, Kenji Maekawa, Norio Sogawa, Shigeo Kitayama, Yoshizo Matsuka, Glenn T Clark, Takuo Kuboki

Journal of prosthodontic research 58 ( 4 ) 217 - 22 2014年10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：ELSEVIER IRELAND LTD

PURPOSE: The aim of this study was to evaluate the correlation between sleep bruxism (SB) frequency and serotonin transporter (SERT)-driven serotonin (5-HT)-uptake in platelets. METHODS: Subjects were dental trainee residents and faculty members of Okayama University Hospital who were aware of having severe or no SB. SB frequency was assessed for 3-consecutive nights by a self-contained electromyographic detector/analyzer, which indicated individual SB levels as one of four grades (score 0, 1, 2 and 3). Subjects were classified as normal control (NC) when SB scores indicated only 0 or 1 during the 3 nights, or as severe SB for scores 2 or 3. Those subjects whose scores fluctuated from 0 to 3 during the 3 nights were omitted from further analysis. Fasting peripheral venous blood samples were collected in the morning following the final SB assessment. Amounts of SERTs proteins collected from peripheral platelets were quantified using ELISA, and SERTs transport activity was assessed by uptake assay using [3H]-5-HT. RESULTS: Thirteen severe SB subjects and 7 NC subjects were eligible. Gender distribution, mean age, 5-HT concentration and total amounts of SERT protein in platelets showed no significant differences between NC and severe SB (p=0.85: Chi-squared test; p=0.64, 0.26, 0.46: t-test). However, [3H]-5-HT uptake by platelets was significantly greater in NC compared to severe SB subjects (12.79±1.97, 8.27±1.91 fmol/10(5) platelets/min, p<0.001, t-test). CONCLUSION: The results of this pilot study suggest a possible correlation between peripheral platelet serotonin transporter uptake ability and SB severity.

DOI： 10.1016/j.jpor.2014.06.003

Dentin-smear remains at self-etch adhesive interface. 国際誌

Atsushi Mine, Jan De Munck, Marcio Vivan Cardoso, Kirsten L Van Landuyt, André Poitevin, Annelies Van Ende, Mariko Matsumoto, Yasuhiro Yoshida, Takuo Kuboki, Hirofumi Yatani, Bart Van Meerbeek

Dental materials : official publication of the Academy of Dental Materials 30 ( 10 ) 1147 - 53 2014年10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：ELSEVIER SCI LTD

OBJECTIVE: The bonding potential of 'mild' self-etch adhesives may be compromised due to smear interference, as they may not dissolve/penetrate the smear layer effectively due to their relatively low acidity. We observed that the thickness of the dentin smear layer differed depending on the surface-preparation methodology used. METHODS: The interaction of an (ultra-)mild self-etch adhesive (Clearfil S3 Bond, Kuraray Noritake) with human dentin, prepared either using a medium-grit diamond bur ('thick', clinically relevant smear layer) or 600-grit SiC-paper ('thin' smear layer), or just fractured (smear-free), was evaluated using high-resolution transmission electron microscopy (TEM). Non-demineralized/demineralized 30-100nm interfacial cross-sections were prepared following common TEM-specimen processing and diamond-knife ultra-microtomy. RESULTS: The adhesive did not dissolve the bur-cut, nor the SiC-ground smear layer, but impregnated it. Within this 'resin-smear complex', hydroxyapatite was abundantly present. At fractured dentin, this complex was not present, while the actual layer of interaction of the adhesive was limited to about 100nm. Non-demineralized 'ultra-thin' (30-50nm) sections confirmed the interfacial ultra-structure to differ for the three surface-preparation methods. An electron dense band was consistently disclosed at the adhesive interface, most likely representing the documented chemical interaction of the functional monomer 10-MDP with Ca. SIGNIFICANCE: The dentin surface-preparation method significantly affects the nature of the smear layer and the interaction with the ultra-mild self-etch adhesive.

DOI： 10.1016/j.dental.2014.07.006

完全な臓器としての歯根膜付きインプラントの開発成功国際誌

Scientific Reports 4 6044 - 6044 2014年8月

簡易貼付型睡眠時ブラキシズム測定装置の測定精度の検討

三木春奈, 水口一, 上枝麻友, 重本修伺, 鈴木善貴, 前川賢治, 松香芳三, 窪木拓男

日本顎関節学会雑誌 26 ( Suppl. ) 125 - 125 2014年7月

Direct interaction between CCN family protein 2 and fibroblast growth factor 1. 国際誌

Tarek Abd El Kader, Satoshi Kubota, Ken Anno, Saho Tanaka, Takashi Nishida, Takayuki Furumatsu, Eriko Aoyama, Takuo Kuboki, Masaharu Takigawa

Journal of cell communication and signaling 8 ( 2 ) 157 - 63 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：SPRINGER

In an attempt to find out a new molecular counterpart of CCN family protein 2 (CCN2), a matricellular protein with multiple functions, we performed an interactome analysis and found fibroblast growth factor (FGF) -1 as one of the candidates. Solid-phase binding assay indicated specific binding between CCN2 and FGF-1. This binding was also confirmed by surface plasmon resonance (SPR) analysis that revealed a dissociation constant (Kd) of 3.98 nM indicating strong molecular interaction between the two. RNA analysis suggested that both FGF-1 and CCN2 could be produced by chondrocytes and thus their interaction in the cartilage is possible. These findings for the first time indicate the direct interaction of CCN2 and FGF-1 and suggest the co-presence of these molecules in the cartilage microenvironment. CCN2 is a well-known promoter of cartilage development and regeneration, whereas the physiological and pathological role of FGF-1 in cartilage mostly remains unclear. Biological role of FGF-1 itself in cartilage is also suspected.

DOI： 10.1007/s12079-014-0232-z

A short-term treatment with tumor necrosis factor-alpha enhances stem cell phenotype of human dental pulp cells. 国際誌

Mayu Ueda, Takuo Fujisawa, Mitsuaki Ono, Emilio Satoshi Hara, Hai Thanh Pham, Ryu Nakajima, Wataru Sonoyama, Takuo Kuboki

Stem cell research & therapy 5 ( 1 ) 31 - 31 2014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：BIOMED CENTRAL LTD

INTRODUCTION: During normal pulp tissue healing, inflammatory cytokines, such as tumor necrosis factor-alpha (TNF-α) or interleukins, act in the initial 48 hours (inflammatory phase) and play important roles not only as chemo-attractants of inflammatory cells and stem/progenitor cells but also in inducing a cascade of reactions toward tissue regeneration or reparative dentin formation or both. Previous reports have shown that inflammatory cytokines regulate the differentiation capacity of dental pulp stem/progenitor cells (DPCs), but none has interrogated the impact of these cytokines on the stem cell phenotype of stem/progenitor cells. This study investigated the effects of a short-term treatment with TNF-α on the stem cell phenotype and differentiation ability of human DPCs. METHODS: An in vivo mouse model of pulp exposure was performed for analysis of expression of the mesenchymal stem cell marker CD146 in DPCs during the initial stage of inflammatory response. For in vitro studies, human DPCs were isolated and incubated with TNF-α for 2 days and passaged to eliminate TNF-α completely. Analysis of stem cell phenotype was performed by quantification of cells positive for mesenchymal stem cell markers SSEA-4 (stage-specific embryonic antigen 4) and CD146 by flow cytometry as well as by quantitative analysis of telomerase activity and mRNA levels of OCT-4 and NANOG. Cell migration, colony-forming ability, and differentiation toward odontogenesis and adipogenesis were also investigated. RESULTS: The pulp exposure model revealed a strong staining for CD146 during the initial inflammatory response, at 2 days after pulp exposure. In vitro experiments demonstrated that a short-term (2-day) treatment of TNF-α increased by twofold the percentage of SSEA-4+ cells. Accordingly, STRO-1, CD146, and SSEA-4 protein levels as well as OCT-4 and NANOG mRNA levels were also significantly upregulated upon TNF-α treatment. A short-term TNF-α treatment also enhanced DPC function, including the ability to form cell colonies, to migrate, and to differentiate into odontogenic and adipogenic lineages. CONCLUSIONS: A short-term treatment with TNF-α enhanced the stem cell phenotype, migration, and differentiation ability of DPCs.

DOI： 10.1186/scrt420

石膏系三次元プリンターにより作製されたコンピュータ骨造形モデルの臨床活用

抜歯窩から採取した未分化間葉系幹細胞

Journal of Dental Research 93 ( 11 ) 1133 - 1140 2014年

Efficient bone formation in swine socket-lift model using E. coli-derived rhBMP-2 adsorbed in b-TCP.

Ono M, Sonoyama W, Yamamoto K, Oida Y, Akiyama K, Shinkawa S, Nakajima R, Pham HT, Hara ES, Kuboki T

199 ( 4 ) 249 - 255 2014年

Efficient bone formation in a swine socket lift model using Escherichia coli-derived recombinant human bone morphogenetic protein-2 adsorbed in β-tricalcium phosphate. 査読国際誌

Ono M, Sonoyama W, Yamamoto K, Oida Y, Akiyama K, Shinkawa S, Nakajima R, Pham HT, Hara ES, Kuboki T

Cells, tissues, organs 199 ( 4 ) 249 - 55 2014年

Immunotherapy in Transplantation 査読

Kentaro Akiyama, Emilio Hara Satoshi, Takuo Kuboki

Stem Cell Biology and Tissue Engineering in Dental Sciences 831 2014年

miRNA-720 regulates the stem cell phenotype and differentiation of human dental pulp-derived mesenchymal stromal cells 査読

Hara Emilio Satoshi, Ono Mitsuaki, Eguchi Takanori, Hai Thanh Pham, Tajima Shoji, Calderwood Stuart K, Matsumoto Takuya, Kuboki Takuo, IEEE

2014 International Symposium on Micro-Nanomechatronics and Human Science (Mhs) 2014年

吉本彰夫, 十河基文, 荒川光, 植村修作, 窪木拓男

日本口腔インプラント学会誌 26 ( 4 ) 723 - 730 2013年12月

記述言語：日本語出版者・発行元：(公社)日本口腔インプラント学会

2名の患者のCTデータを基に、石膏系の三次元プリンターによるラピッドプロトタイピングのコンピュータ骨造形モデルを作製し、その臨床活用法を検討した。上顎右側洞底挙上術を行う患者では、術前診査において患者固有の上顎洞の解剖学的特徴を把握でき、CT画像による骨内の動脈走行に注意を払いつつ、上顎洞底挙上術の開窓位置や開窓範囲を決定する一助となった。また、上顎無歯顎のオープンフラップ施術後に即時負荷を与える患者では、顎骨データと全部床義歯の咬合面データを連結した一体のデータを作成することで骨造形モデルでも咬合を確立でき、事前に即時負荷用プロビジョナルレストレーションを作製できた。CTデータの新しい活用法の一つとして、石膏系の三次元プリンターを用いたコンピュータ骨造形モデルの活用は非常に有益な方法であることがわかった。

J-GLOBAL

Hydrolytic stability of three-step etch-and-rinse adhesives in occlusal class-I cavities. 査読国際誌

Jan De Munck, Atsushi Mine, Marcio Vivan Cardoso, Kirsten L Van Landuyt, Anne-Katrin Lührs, André Poitevin, Masao Hanabusa, Takuo Kuboki, Bart Van Meerbeek

Clinical oral investigations 17 ( 8 ) 1911 - 8 2013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：8

OBJECTIVES: A dental adhesive without small and hydrophilic monomers such as 2-hydroxyethyl methacrylate (HEMA) and triethylene glycol dimethacrylate (TEGDMA) would be beneficial in order to avoid contact allergies. However, these monomers are important to increase infiltration and polymerization of the adhesive. Therefore, the purpose of this study was to evaluate the bonding effectiveness and bond durability of a more hydrophobic and biocompatible adhesive as compared to a conventional three-step etch-and-rinse adhesive. METHODS: Sixteen non-carious human third molars were used to determine the micro-tensile bond strength testing (μTBS) and interfacial ultrastructure by transmission electron microscopy (TEM) of the more hydrophobic cmf adhesive system (Saremco) adhesive as compared to the control OptiBond FL (Kerr). RESULTS: The more hydrophobic and biocompatible three-step etch-and-rinse adhesive was able to produce a reasonable short-time bonding effectiveness. In the long term, the collagen fibrils in the hybrid layer were not effectively protected and were prone to hydrolytic degradation. As a result, long-term bonding effectiveness of this novel adhesive was very low. CONCLUSIONS: Application of a more hydrophobic adhesive without altering the application procedure considerably results in a reduced durability of the created bond CLINICAL RELEVANCE: Omitting small and hydrophilic components from the adhesive formulation may impair the durability of your composite restoration.

DOI： 10.1007/s00784-012-0884-0

補綴歯科治療病名システムの信頼性と妥当性の検討査読

松香芳三, 萩原芳幸, 玉置勝司, 竹内久裕, 藤澤政紀, 小野高裕, 築山能大, 永尾寛, 津賀一弘, 會田英紀, 近藤尚知, 笛木賢治, 塚崎弘明, 石橋寛二, 藤井重壽, 平井敏博, 佐々木啓一, 矢谷博文, 五十嵐順正, 佐藤裕二, 市川哲雄, 松村英雄, 山森徹雄, 窪木拓男, 馬場一美, 古谷野潔

日補綴会誌 5 ( 3 ) 281 - 290 2013年11月

記述言語：日本語出版者・発行元：(公社)日本補綴歯科学会

目的:(社)日本補綴歯科学会は病態とその発現機序の把握に基づく適切な補綴歯科治療を国民に提供するために、補綴歯科治療における新たな病名システムを提案した。これは患者に生じている「障害」を病名の基本とし、この障害を引き起こしている「要因」を併記して病名システムとするものであり、「A(要因)によるB(障害)」を病名システムの基本的な表現法としている。本研究の目的は考案した方法に従って決定した補綴歯科治療における病名の信頼性と妥当性を検討することである。方法:模擬患者カルテを作成し、(社)日本補綴歯科学会診療ガイドライン委員会で模範解答としての病名(以下、模範病名)を決定した。その後、合計50名の評価者(日本補綴歯科学会専門医(以下、補綴歯科専門医)ならびに大学病院研修歯科医(以下、研修医))に診断をしてもらい、評価者間における病名の一致度(信頼性)ならびに(社)日本補綴歯科学会診療ガイドライン委員会による模範病名との一致度(妥当性)を検討した。結果:評価者間の一致度を検討するための算出したKrippendorff's αは全体では0.378、補綴歯科専門医では0.370、研修医では0.401であった。Krippendorff's αは模範病名との一致度の高い上位10名の評価者(補綴歯科専門医:3名、研修医:7名)では0.524、上位2名の評価者(補綴歯科専門医:1名、研修医:1名)では0.648と上昇した。日常的に頻繁に遭遇する病名に関しては模範病名との一致度が高かったが、日常的に遭遇しない病名は模範病名との一致度は低い状況であった。さらに、模範病名との一致度とアンケート回答時間や診療経験年数の関連性を検討したところ、相関関係はみられなかった。結論:全評価者間の一致度を指標とした本病名システムの信頼性は高くはなかったが、模範病名との一致度の高い評価者間では一致度が高かった。日常的に遭遇する補綴関連病名については模範病名との一致度が高かった。以上から(公社)日本補綴歯科学会の新しい病名システムは臨床上十分な信頼性と妥当性を有することが示唆された。(著者抄録)

DOI： 10.2186/ajps.5.281

J-GLOBAL

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Regulation of CCN2 gene expression and possible roles in developing tooth germs. 査読国際誌

Manabu Kanyama, Tsuyoshi Shimo, Hiroki Sugito, Motohiko Nagayama, Takuo Kuboki, Maurizio Pacifici, Eiki Koyama

Archives of oral biology 58 ( 11 ) 1659 - 66 2013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

CCN proteins are extracellular and cell-associated molecules involved in several developmental processes, but their expression patterns and regulation in tooth development remain unclear. Here we first determined the expression patterns of CCN genes in mouse tooth germs. We found that at early stages CCN2 was detected in dental lamina, dental mesenchyme, and primary enamel knot, while other CCN family members were expressed broadly. By the bell stage, all members were expressed in differentiating odontoblasts and ameloblasts, but CCN1 and CCN2 transcripts were conspicuous in differentiating osteoblasts in dental follicle. Next, we asked what signalling molecules regulate CCN2 expression and what roles CCN2 may have. We found that upon surgical removal of dental epithelium CCN2 was not longer expressed in dental mesenchyme in cultured bud stage germs. Implantation of beads pre-coated with BMPs and FGFs onto E12-13 mandibular explants induced CCN2 expression in dental mesenchyme. There was a dose-dependent effect of BMP-4 on CCN2 induction; a concentration of 100 ng/μl was able to induce strong CCN2 expression while a minimum concentration of 25 ng/μl was needed to elicit appreciable expression. Importantly, Noggin treatment inhibited endogenous and BMP-induced CCN2 expression, verifying that CCN2 expression in developing tooth germs requires BMP signalling. Lastly, we found that rCCN2 stimulated proliferation in dental mesenchyme in a dose-dependent manner. Together, the data indicate that expression of CCN genes is spatio-temporally regulated in developing tooth germs. CCN2 expression appears to depend on epithelial and mesenchymal-derived signalling factors, and CCN2 can elicit strong proliferation in dental mesenchyme.

DOI： 10.1016/j.archoralbio.2013.08.010