Faculty Profiles - KUBOKI Takuo

Papers - KUBOKI Takuo

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Simvastatin induces the odontogenic differentiation of human dental pulp stem cells in vitro and in vivo. Reviewed

Yosuke Okamoto, Wataru Sonoyama, Mitsuaki Ono, Kentaro Akiyama, Takuo Fujisawa, Masamitsu Oshima, Yohei Tsuchimoto, Yoshizo Matsuka, Tatsuji Yasuda, Songtao Shi, Takuo Kuboki

Journal of Endodontics Vol.35 ( No.3 ) 367 - 372 2009.3

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

Statin, 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor, is known to promote bone formation. However, it is not clear whether statin affects the differentiation of pulp cells. This study used a cell proliferation assay, cell cycle analysis, quantitative reverse transcriptase polymerase chain reaction (RT-PCR) and in vivo transplantation to examine the effects of simvastatin on human dental pulp stem cells (DPSCs) in vitro and in vivo. Simvastatin at 1 mumol/L was able to significantly suppress the proliferation of DPSCs without inducing apoptosis. Quantitative RT-PCR revealed both osteocalcin and dentin sialophosphoprotein to be significantly up-regulated when DPSCs were cultured with simvastatin in comparison to bone morphogenetic protein-2 treatment. The in vivo transplantation data showed that simvastatin treatment promoted mineralized tissue formation. Taken together, these results suggest that statin might be an ideal active ingredient to accelerate the differentiation of DPSCs.

DOI： 10.1016/j.joen.2008.11.024

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Corrigendum to "botulinum toxin type A (150 kDa) decreases exaggerated neurotransmitter release from trigeminal ganglion neurons and relieves neuropathy behaviors induced by infraorbital nerve constriction" [Neuroscience 159 (2009) 1422-1429] (DOI:10.1016 Reviewed

Kitamura Y, Matsuka Y, Spigelman I, Ishihara Y, Yamamoto Y, Sonoyama W, Kamioka H, Yamashiro T, Kuboki T, Oguma K

Neuroscience 161 ( 3 ) 2009

Botulinum toxin blocks neurotransmitter release from somata of rat neuropathic trigeminal ganglion neurons Reviewed

Kitamura Yoichi, Matsuka Yoshizo, Ishihara Yoshihito, Spigelman Igor, Yamamoto Yumiko, Sonoyama Wataru, Kamioka Horoshi, Yamashiro Takashi, Oguma Keiji, Kuboki Takuo

TOXICON 51 6 - 6 2008.6

Promotion of hydroxyapatite-associated, stem cell-based bone regeneration by CCN2. Reviewed International journal

Mitsuaki Ono, Satoshi Kubota, Takuo Fujisawa, Wataru Sonoyama, Harumi Kawaki, Kentaro Akiyama, Kengo Shimono, Masamitsu Oshima, Takashi Nishida, Yasuhiro Yoshida, Kazuomi Suzuki, Masaharu Takigawa, Takuo Kuboki

Cell transplantation 17 ( 1-2 ) 231 - 40 2008

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

Multiple roles have been already recognized for CCN2 in cartilage development and regeneration. However, the effects of CCN2 on bone regeneration remain to be elucidated. In this study, the utility of CCN2 on bone regeneration was examined in vitro and in vivo in combination with hydroxyapatite (HAp) as a scaffold. Human bone marrow stromal cells (hBMSCs) were isolated from human iliac bone marrow aspirates of healthy donors and expanded, and the effects of CCN2 on their proliferation and migration were examined in vitro. The proliferation of hBMSCs on a plastic or HAp plate was significantly enhanced by CCN2. Moreover, the migration of hBMSCs also dramatically increased by CCN2. Interestingly, a C-terminal signal modular fragment of CCN2 (CT-module) also enhanced the cell proliferation and migration as efficiently as the full-length CCN2. Next, in order to estimate the effect of CCN2 on the migration and survival of hBMSCs and bone formation inside the HAp scaffold in vivo, two experiments were performed. First, the porous HAp carrier was cultured with hBMSCs for a week, and the cell-scaffold hybrid was transplanted with or without CCN2 subcutaneously into immunocompromised mice. CCN2 accelerated the hBMSC-like cell migration and survival inside the porous HAp within 4 weeks after transplantation. Second, the porous HAp carrier with or without CCN2 was directly implanted into bone defects within a rabbit mandible, and bone regeneration inside was evaluated. As a result, CCN2 efficiently induced the cell invasion and bone formation inside the porous HAp scaffold. These findings suggest that CCN2 and its CT-module fragment could be useful for regeneration and reconstruction of large-scale bone defects.

DOI： 10.3727/000000008783907143

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粘膜圧迫により疼痛の軽減がみられた三叉神経に生じた持続性ニューロパシー性疼痛の2例 Reviewed

Yoshizo Matsuka, 熊田愛, 前川賢治, 水口一, 窪木拓男

Japanese Journal of Orofacial Pain Vol.1 ( No.1 ) 55 - 59 2008

われわれの行っているインプラント治療の臨床的検討

高木愼, 水川展吉, 福永城司, 菅原利夫, 完山学, 窪木拓男, 丸尾幸憲, 皆木省吾

岡山歯学会雑誌 26 ( 1 ) 49 - 50 2007.6

骨芽細胞様細胞(MC3T3E1)分化に対するチタンの影響

秋山謙太郎, 藤澤拓生, 明貝文夫, 大野充明, 吉田靖弘, 高柴正悟, 鈴木一臣, 窪木拓男

日本骨代謝学会学術集会プログラム抄録集 24回 229 - 229 2006.7

Effect of surface pre-treatment on durability of resin-based cements bonded to titanium. International journal

Youhei Tsuchimoto, Yasuhiro Yoshida, Maho Takeuchi, Atsushi Mine, Hirofumi Yatani, Yoh-ichi Tagawa, Bart Van Meerbeek, Kazuomi Suzuki, Takuo Kuboki

Dental materials : official publication of the Academy of Dental Materials 22 ( 6 ) 545 - 52 2006.6

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

OBJECTIVES: Titanium has in recent years become a material of great opportunities in dentistry and orthopedics. In this study, we searched for a method to effectively pre-treat titanium as part of an adhesive luting and/or resin-veneering procedure. METHODS: Ti plates were treated with two different solutions of hydrochloric acid (0.1, 1N), 37wt% phosphoric acid or kept untreated (control). The tensile bond strength of the composite cement (Panavia Fluoro Cement, Kuraray) to the untreated and pre-treated Ti plates was determined without and after 20,000 thermo-cycles. XPS was used to chemically analyze the effect of the three Ti pre-treatments, as well as the interaction of 10-methacryloxydecyl dihydrogen phosphate (10-MDP; functional monomer of Panavia Fluoro Cement) with Ti treated with 1N HCl. RESULTS: Although no significant difference in immediate tensile bond strength was measured, thermo-cycling significantly decreased the bond strength of all experimental groups except for Ti treated with 1N HCl. No difference in surface roughness was found between untreated and treated Ti. XPS demonstrated that HCl effectively decontaminated Ti in a dose-dependent manner, while H(3)PO(4) was strongly adsorbed on the Ti surface. While the latter potentially inhibits adsorption of the phosphoric groups of the functional monomer 10-MDP, Ti pre-treated with 1N HCl improved the adsorption of 10-MDP as compared to untreated Ti. SIGNIFICANCE: It is concluded that 1N HCl effectively pre-treats Ti in order to make it more receptive to resin, while H(3)PO(4) should better not be used.

DOI： 10.1016/j.dental.2005.08.002

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骨芽細胞様細胞株(MC3T3E1細胞)の細胞接着,増殖,分化および遺伝子発現に対するチタンの影響

秋山謙太郎, 藤澤拓生, 明貝文夫, 完山学, 吉田靖弘, 高柴正悟, 鈴木一臣, 窪木拓男

日本口腔インプラント学会誌 19 ( 1 ) 32 - 32 2006.3

岡山大学歯学部1年次生におけるテュートリアル教育の問題点と今後の課題

完山学, 久保田聡, 市川博之, 十川紀夫, 平田あずみ, 美藤純弘, 岡本信, 福永智広, 香川智正, 中山周子, 坂本友紀, 吉田登志子, 宮脇卓也, 西村英紀, 松村誠士, 窪木拓男

岡山歯学会雑誌 24 ( 2 ) 97 - 104 2005.12

[Longitudinal follow-up of symptom severity of a patient with continuous neuropathic pain assessed by actual drug consumption record].

Chiyomi Nagamatsu, Kumiko Nawachi, Atsushi Mine, Kenji Maekawa, Takuo Kuboki

Nihon Hotetsu Shika Gakkai zasshi 49 ( 5 ) 691 - 6 2005.10

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

PATIENT: The patient was a 56-year-old female, suffering from severe burning pain in the left maxillary alveolar area. The pain was initiated by periodontal curettage under no local anesthesia. No abnormal local findings in dental X-ray images and no abnormal central nervous system findings in brain magnetic resonance images were observed. Extraction of the tooth temporarily abated her pain, however the pain recurred gradually. Since the observed signs and symptoms of the patient agreed well with the diagnostic standard for continuous neuropathic pain (atypical alveolar pain) of Graff-Radford & Solberg (1992), medications (tricyclic antidepressant, non-steroidal anti-inflammatory drug (NSAID), anxiolytics, topical application of capsacin cream) and supportive psychological therapy have been administered continuously. To measure symptom severity over a long term, daily amounts of taken NSAID and anxiolytics were recorded continuously by the patient in a simple table diary for 7 years. As a result, the total amount of the drugs per month gradually decreased with the passage of time. DISCUSSION AND CONCLUSIONS: Clinical evidence regarding the prognosis of continuous neuropathic pain in the orofacial region is obviously insufficient, making it difficult to inform such patients of their prognosis of the disorder on the basis of clinical evidence. In this report, we have plotted the total amount of drugs taken per month by the patient as a good reference for the pain severity and the results clearly suggest that pain severity decreased with time. This kind of information may help both clinicians and patients for case-based explanation of the disorder.

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Adsorption of polyphosphoric acid to titanium surface and its effect on hBMSC attachment Reviewed

K. Maekawa, A. Mine, T. Fujisawa, Y. Yoshida, K. Suzuki, T. Kuboki

International Congress Series 1284 332 - 333 2005.9

[Needs and current research directions of biological regenerative medicine in prosthodontic practice--to attain reliable and sophisticated dental implant therapy].

Takuo Kuboki

Nihon Hotetsu Shika Gakkai zasshi 49 ( 4 ) 576 - 86 2005.8

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

Appropriate dental implants are known to improve the quality of life in totally and partially edentulous patients, as well as to prevent future tooth loss, and so prosthetic treatment modalities have drastically changed recently with reconstruction therapy. However, dental implant therapy did not win the confidence of the dental community in the early developmental stage, until osseointegration between titanium and surrounding bone tissue was discovered and the modality utilizing osseointegration became reliable and produced durable treatment outcomes for long-term function. On the other hand, the biological mechanism of osseointegration has not been clarified yet and the time required for osseointegration is still long, e.g. three to four months. As well, when the implant is applied in the upper posterior region, the acquisition of osseointegration and the long-term survival of the implant are still not clinically adequate. Therefore, to reduce the time required for osseointegration and to regenerate enough alveolar bone mass in the target implantation site, the oral implant modality must be made more useful and potent as one of the treatment options for partial and total edentulism. With this background, we have studied biological strategies for reducing the time required for osseointegration and for regenerating enough alveolar bone mass, e.g., investigation of the specific genes for osseointegration between titanium and bone, nano-level surface modification of the titanium, biodegradable apatite foam for alveolar bone regeneration, application of bone formation-related growth factors with biological scaffold, and autologous cell transplantation of bone marrow derived mesenchymal stem cells. In this article, we review the current status of regenerative medicine being applied in prosthodontics and discuss our future research direction.

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交通事故外傷による顎顔面の変形と咬合異常を呈した症例の一治験例

藤山光治, 山城隆, 藤井昭仁, 窪木拓男, 山本照子

日本矯正歯科学会大会プログラム・抄録集 63回 301 - 301 2004.11

再生医療と歯科補綴学の接点補綴治療における再生医療のニーズと現時点での研究動向確実かつ質の高いインプラント治療をめざして

窪木拓男

日本補綴歯科学会雑誌 48 ( 112回特別 ) 41 - 41 2004.10

Regeneration of defects in articular cartilage in rat knee joints by CCN2 (connective tissue growth factor). International journal

Takashi Nishida, Satoshi Kubota, Shunji Kojima, Takuo Kuboki, Kyouji Nakao, Toshihiro Kushibiki, Yasuhiko Tabata, Masaharu Takigawa

Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research 19 ( 8 ) 1308 - 19 2004.8

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

UNLABELLED: CTGF/CCN2, a hypertrophic chondrocyte-specific gene product, possessed the ability to repair damaged articular cartilage in two animal models, which were experimental osteoarthritis and full-thickness defects of articular cartilage. These findings suggest that CTGF/CCN2 may be useful in regeneration of articular cartilage. INTRODUCTION: Connective tissue growth factor (CTGF)/CCN2 is a unique growth factor that stimulates the proliferation and differentiation, but not hypertrophy, of articular chondrocytes in vitro. The objective of this study was to investigate the therapeutic use of CTGF/CCN2. MATERIALS AND METHODS: The effects of recombinant CTGF/CCN2 (rCTGF/CCN2) on repair of damaged cartilage were evaluated by using both the monoiodoacetic acid (MIA)-induced experimental rat osteoarthritis (OA) model and full-thickness defects of rat articular cartilage in vivo. RESULTS: In the MIA-induced OA model, quantitative real-time RT-PCR assays showed a significant increase in the level of CTGF/CCN2 mRNA, and immunohistochemical analysis and in situ hybridization revealed that the clustered chondrocytes, in which clustering indicates an attempt to repair the damaged cartilage, produced CTGF/CCN2. Therefore, CTGF/CCN2 was suspected to play critical roles in cartilage repair. In fact, a single injection of rCTGF/CCN2 incorporated in gelatin hydrogel (rCTGF/CCN2-hydrogel) into the joint cavity of MIA-induced OA model rats repaired their articular cartilage to the extent that it became histologically similar to normal articular cartilage. Next, to examine the effect of rCTGF/CCN2 on the repair of articular cartilage, we created defects (2 mm in diameter) on the surface of articular cartilage in situ and implanted rCTGF/CCN2-hydrogel or PBS-hydrogel therein with collagen sponge. In the group implanted with rCTGF/CCN2-hydrogel collagen, new cartilage filled the defect 4 weeks postoperatively. In contrast, only soft tissue repair occurred when the PBS-hydrogel collagen was implanted. Consistent with these in vivo effects, rCTGF/CCN2 enhanced type II collagen and aggrecan mRNA expression in mouse bone marrow-derived stromal cells and induced chondrogenesis in vitro. CONCLUSION: These findings suggest the utility of CTGF/CCN2 in the regeneration of articular cartilage.

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[Cartilage and mechanical stress from the point of the view of development, growth, pathology and therapeutic aspects].

Takuo Fujisawa, Masaharu Takigawa, Takuo Kuboki

Clinical calcium 14 ( 7 ) 29 - 35 2004.7

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

Articular cartilage is always subjected to various types, magnitudes and cycles of mechanical stress. While it has been recognized that these stresses regulate the chondrocyte growth and differentiation, the mechanism is still unclear. Here, we summarize the effect of mechanical stress on cartilage metabolism from the point of view of development, growth, pathology and therapeutic aspect.

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掌蹠膿疱症を合併した重度歯周炎患者に対して行っている歯科的アプローチの一例

赤澤洋, 西村英紀, 前田博史, 峯篤史, 前川賢治, 窪木拓男, 高柴正悟

岡山歯学会雑誌 23 ( 1 ) 190 - 190 2004.6

Connective tissue growth factor expressed in rat alveolar bone regeneration sites after tooth extraction. International journal

Manabu Kanyama, Takuo Kuboki, Kentaro Akiyama, Kumiko Nawachi, Fabiora M Miyauchi, Hirofumi Yatani, Satoshi Kubota, Tohru Nakanishi, Masaharu Takigawa

Archives of oral biology 48 ( 10 ) 723 - 30 2003.10

Language：English Publishing type：Research paper (scientific journal) Publisher：PERGAMON-ELSEVIER SCIENCE LTD

OBJECTIVE: To understand bone regeneration process after tooth extraction could be a clue to develop a new strategy for alveolar bone reconstruction. Recently, accumulated evidences support that connective tissue growth factor (CTGF) is implicated in tissue repair of many tissues. In this study, we investigated the spatial and temporal expression of CTGF in the rat tooth extraction sockets. DESIGN: Five weeks old wild type male rats (weighing 120 g) were used for this experiment. Expression of CTGF was determined by immunohistochemistry and in situ hybridization in the rat upper molar tooth extraction sockets at 2, 4, 7, 10 and 14 days after tooth extraction. RESULTS: CTGF was expressed strongly in the endothelial cells migrating into the granulation tissue at the bottom of the sockets during 4 days after tooth extraction. During the reparative process, no apparent chondrocyte-like cell appeared in the sockets, while osteoblast-like cells proliferated in the sockets with low CTGF expression at 7, 10, 14 days after extraction. As expected, no staining was observed with the preimmune rabbit IgG and CTGF sense probe. CTGF may play an important role in angiogenesis and granulation tissue formation specifically at early healing stage after tooth extraction to initiate alveolar bone repair. CONCLUSION: CTGF was expressed at early healing stage of the rat tooth extraction wound.

DOI： 10.1016/S0003-9969(03)00153-5

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A repetitive, steady mouth opening induced an osteoarthritis-like lesion in the rabbit temporomandibular joint

T. Fujisawa, T. Kuboki, T. Kasai, W. Sonoyama, S. Kojima, J. Uehara, C. Komori, H. Yatani, T. Hattori, M. Takigawa

Journal of Dental Research 82 ( 9 ) 731 - 735 2003

Language：English Publishing type：Research paper (scientific journal) Publisher：Intern. and American Associations for Dental Research

Although excessive mechanical stress is assumed to be one of the factors contributing to pathogenesis of temporomandibular joint (TMJ) osteoarthritis (OA), no pure mechanical-stress-induced OA model has been developed without surgical manipulation or puncture of the joint cavity. The purpose of this study was to establish a genuine mechanical-stress-induced OA model of the rabbit TMJ. In the experimental rabbits, repetitive, forced jaw-opening, 3 hrs/day for 5 days, was applied with the use of a general anesthesia protocol. By histological assessment of the TMJ articular tissues, partial eburnation of the articular cartilage, reactive marginal proliferation of the articular cartilage chondrocytes, and nested proliferation of chondrocytes in the subchondral bone area were observed at 7 days after the repetitive, forced-jaw-opening period. These results suggest that the repetitive, forced-jaw-opening protocol without surgical intervention can induce evident OA-like lesions in the rabbit TMJ, and this OA model may greatly contribute to the elucidation of the cartilage degradation mechanism in TMJ OA.

DOI： 10.1177/154405910308200914

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