Faculty Profiles - KUBOKI Takuo

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 2014.10

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

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 2014.10

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

Functional tooth restoration by next-generation bio-hybrid implant as a bio-hybrid artificial organ replacement therapy. International journal

Masamitsu Oshima, Kaoru Inoue, Kei Nakajima, Tetsuhiko Tachikawa, Hiromichi Yamazaki, Tomohide Isobe, Ayaka Sugawara, Miho Ogawa, Chie Tanaka, Masahiro Saito, Shohei Kasugai, Teruko Takano-Yamamoto, Takashi Inoue, Katsunari Tezuka, Takuo Kuboki, Akira Yamaguchi, Takashi Tsuji

Scientific reports 4 6044 - 6044 2014.8

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

Bio-hybrid artificial organs are an attractive concept to restore organ function through precise biological cooperation with surrounding tissues in vivo. However, in bio-hybrid artificial organs, an artificial organ with fibrous connective tissues, including muscles, tendons and ligaments, has not been developed. Here, we have enveloped with embryonic dental follicle tissue around a HA-coated dental implant, and transplanted into the lower first molar region of a murine tooth-loss model. We successfully developed a novel fibrous connected tooth implant using a HA-coated dental implant and dental follicle stem cells as a bio-hybrid organ. This bio-hybrid implant restored physiological functions, including bone remodelling, regeneration of severe bone-defect and responsiveness to noxious stimuli, through regeneration with periodontal tissues, such as periodontal ligament and cementum. Thus, this study represents the potential for a next-generation bio-hybrid implant for tooth loss as a future bio-hybrid artificial organ replacement therapy.

DOI： 10.1038/srep06044

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 2014.7

Direct interaction between CCN family protein 2 and fibroblast growth factor 1. International journal

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 2014.6

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

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 2014.2

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

Mesenchymal Stem/Progenitor Cell Isolation from Tooth Extraction Sockets

Nakajima, R., Ono, M., Hara, E. S., Oida, Y., Shinkawa, S., Pham, H. T., Akiyama, K., Sonoyama, W., Maekawa, K., Kuboki, T.

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

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

Efficient bone formation in a swine socket lift model using Escherichia coli-derived recombinant human bone morphogenetic protein-2 adsorbed in β-tricalcium phosphate. Reviewed International journal

Mitsuaki Ono, Wataru Sonoyama, Katushi Yamamoto, Yasutaka Oida, Kentaro Akiyama, Shigehiko Shinkawa, Ryu Nakajima, Hai T Pham, Emilio S Hara, Takuo Kuboki

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

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

Several preclinical studies have shown that Escherichia coli-derived bone morphogenetic protein-2 (E-BMP-2) is as effective as mammalian cell-derived bone morphogenetic protein-2 (C-BMP-2) in the treatment of bone defects. However, further investigation of the effectiveness and determination of the optimal dosage of E-BMP-2 in large animals are still necessary before its full application in humans. This study investigated the efficiency of different concentrations of E-BMP-2 adsorbed in β-TCP for bone augmentation and osseointegration of immediate dental implants in a swine socket lift model. Following exposure of the maxillary sinus lateral wall, a 3.4-mm (diameter) cavity was drilled and filled with 0.1 g of β-TCP containing different doses of E-BMP-2 (0, 10, 30, or 100 μg/site) to lift the Schneiderian membrane. A dental implant was then immediately inserted. Bone-to-implant contact (BIC) and bone density (BD) examined via histological analysis were used as parameters to assess E-BMP-2 efficiency in bone formation. The implant stability quotient (ISQ) was measured using Osstell to determine the effect of E-BMP-2/β-TCP on implant stability. After 8 weeks, the groups that received 30 and 100 μg of E-BMP-2 showed substantial new bone formation in the elevated space, while no bone formation was observed with β-TCP alone. Accordingly, BIC and BD presented a dose-dependent response to increasing doses of E-BMP-2. However, there was no increase in implant stability with E-BMP-2 treatment. In conclusion, the E-BMP-2/β-TCP combination was efficient in bone formation and osseointegration of dental implants in a socket lift model in mini-pigs.

DOI： 10.1159/000369061

Immunotherapy in Transplantation Reviewed

Akiyama, K., Hara Satoshi, E., Kuboki, T.

Stem Cell Biology and Tissue Engineering in Dental Sciences 831 2014

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

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

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

In this study we performed a microRNA (miRNA) array to characterize a stem cell population from human dental pulp-derived mesenchymal stromal cells (hDPSCs). miRNA-720 was found to regulate the stem cell phenotype of hDPSCs by targeting the pluripotency factor NANOG.

DOI： 10.1109/MHS.2014.7006080

Clinical Applications of Computerized Bone Models Made by 3D Plaster Printing

吉本彰夫, 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 2013.12

Hydrolytic stability of three-step etch-and-rinse adhesives in occlusal class-I cavities. Reviewed International journal

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 2013.11

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

Reliability and validity of diagnostic nomenclature system of prosthetic dentistry Reviewed

松香芳三, 松香芳三, 萩原芳幸, 萩原芳幸, 玉置勝司, 玉置勝司, 竹内久裕, 藤澤政紀, 藤澤政紀, 小野高裕, 小野高裕, 築山能大, 永尾寛, 津賀一弘, 會田英紀, 會田英紀, 近藤尚知, 近藤尚知, 笛木賢治, 笛木賢治, 塚崎弘明, 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, 馬場一美, 古谷野潔

日本補綴歯科学会誌 5 ( 3 ) 281 - 290 2013.11

Regulation of CCN2 gene expression and possible roles in developing tooth germs. Reviewed International journal

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

Archives of oral biology 58 ( 11 ) 1659 - 66 2013.11

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