Faculty Profiles - KUBOKI Takuo

Anti-fibrotic effect of CCN3 accompanied by altered gene expression profile of the CCN family. Reviewed International journal

Tarek Abd El Kader, Satoshi Kubota, Danilo Janune, Takashi Nishida, Takako Hattori, Eriko Aoyama, Bernard Perbal, Takuo Kuboki, Masaharu Takigawa

Journal of cell communication and signaling 7 ( 1 ) 11 - 8 2013.3

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

CCN family proteins 2 and 3 (CCN2 and CCN3) belong to the CCN family of proteins, all having a high level of structural similarity. It is widely known that CCN2 is a profibrotic molecule that mediates the development of fibrotic disorders in many different tissues and organs. In contrast, CCN3 has been recently suggested to act as an anti-fibrotic factor in several tissues. This CCN3 action was shown earlier to be exerted by the repression of the CCN2 gene expression in kidney tissue, whereas different findings were obtained for liver cells. Thus, the molecular action of CCN3 yielding its anti-fibrotic effect is still controversial. Here, using a general model of fibrosis, we evaluated the effect of CCN3 overexpression on the gene expression of all of the CCN family members, as well as on that of fibrotic marker genes. As a result, repression of CCN2 gene expression was modest, while type I collagen and α-smooth muscle actin gene expression was prominently repressed. Interestingly, not only CCN2, but also CCN4 gene expression showed a decrease upon CCN3 overexpression. These findings indicate that fibrotic gene induction is under the control of a complex molecular network conducted by CCN family members functioning together.

DOI： 10.1007/s12079-012-0180-4

Novel chondrogenic and chondroprotective effects of the natural compound harmine. Reviewed International journal

Emilio Satoshi Hara, Mitsuaki Ono, Satoshi Kubota, Wataru Sonoyama, Yasutaka Oida, Takako Hattori, Takashi Nishida, Takayuki Furumatsu, Toshifumi Ozaki, Masaharu Takigawa, Takuo Kuboki

Biochimie 95 ( 2 ) 374 - 81 2013.2

Language：English Publishing type：Research paper (scientific journal) Publisher：ELSEVIER FRANCE-EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER

A significant number of natural compounds have been shown to regulate the behavior of the cells, in collaboration with cellular proteins. CCN2/connective tissue growth factor (CTGF) has been reported to have essential roles in cartilage development, chondrocyte proliferation and differentiation as well as regulation of the extracellular matrix metabolism. Previous studies demonstrated the capability of CCN2 to regenerate surgical defects in articular cartilage of rat knee. Also, transgenic mice over-expressing cartilage-specific CCN2 were shown to be more resistant to aging-related cartilage degradation. We hypothesized that small molecules that induce CCN2 in chondrocytes could be novel candidates to increase the resistance to aging-related cartilage degradation, or even to correct cartilage degenerative changes incurred in OA. Therefore, this study screened a compound library and identified the β-carboline alkaloid harmine as a novel inducer of CCN2 in human chondrocytic HCS-2/8 cells and osteoarthritic articular chondrocytes. Harmine increased the expression of the cartilage markers aggrecan and COL2α1, as well as that of the master regulator of chondrogenesis, SOX-9. Moreover, harmine notably induced chondrogenesis of prechondrocytic ATDC5 cells in micromass cultures. The chondroprotective effect of harmine was investigated under inflammatory condition by stimulation with TNFα, and harmine was shown to ameliorate TNFα-induced decrease in expression of CCN2 and cartilage markers. These findings uncover novel chondrogenic effects of harmine and indicate harmine as a potential drug for prevention and/or repair of cartilage degradation.

DOI： 10.1016/j.biochi.2012.10.016

Regeneration of calvarial defects with Escherichia coli -derived rhBMP-2 adsorbed in PLGA membrane. Reviewed International journal

Mitsuaki Ono, Wataru Sonoyama, Kazuki Nema, Emilio Satoshi Hara, Yasutaka Oida, Hai Thanh Pham, Katushi Yamamoto, Kazuo Hirota, Kazushige Sugama, Walter Sebald, Takuo Kuboki

Cells, tissues, organs 198 ( 5 ) 367 - 76 2013

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

OBJECTIVE: Escherichia coli-derived recombinant human bone morphogenetic protein-2 (E-BMP-2) has been shown to be as effective as mammalian cell-derived BMP-2. However, several in vitro and in vivo experiments are still necessary to validate the effectiveness of E-BMP-2 due to the difference in synthesis process, mainly related to protein nonglycosylation. The objective of this study was to investigate whether biodegradable polylactide-co-glycolide (PLGA) membrane is a suitable carrier for E-BMP-2 delivery for bone regeneration of critical-sized defects in rat calvaria. MATERIALS AND METHODS: First, the osteoinductive effect of E-BMP-2 was confirmed in vitro in mouse bone marrow stromal cells by analysis of osteocalcin mRNA levels, and calcium deposition was detected by alizarin red staining. Before in vivo experiments, the release profile of E-BMP-2 from PLGA membranes was determined by ELISA. E-BMP-2 (0, 1, 5 and 10 μg/μl) was applied for ectopic and orthotopic bone formation and was analyzed by X-ray, micro-CT and histology. RESULTS: Release-profile testing showed that PLGA membrane could retain 94% of the initially applied E-BMP-2. Ectopic bone formation assay revealed that combination of E-BMP-2/PLGA membrane strongly induced bone formation. Stronger osteoinductivity with complete repair of critical-sized defects was observed only with PLGA membranes adsorbed with 5 and 10 μg/μl of E-BMP-2, whereas no bone formation was observed in the groups that received no membrane or 0-μg/μl dose of E-BMP-2. CONCLUSION: PLGA membrane was shown to be a suitable carrier for sustained release of E-BMP-2, and the E-BMP-2/PLGA membrane combination was demonstrated to be efficient in bone regeneration in a model of critical-sized defects.

DOI： 10.1159/000356947

The CCN2-inducer harmine promotes chondrogenesis and protects against TNFα-induced ablation of chondrocytic phenotype Reviewed

Hara, E.S, M. Ono, S. Kubota, W. Sonoyama, Y. Oida, T. Hattori, T. Nishida, T. Furumatsu, T. Ozaki, M. Takigawa, T. Kuboki

Biochimie 95 374 - 381 2013

A problem-based learning tutorial for dental students regarding elderly residents in a nursing home in Japan. Reviewed International journal

Yoshizo Matsuka, Ryu Nakajima, Haruna Miki, Aya Kimura, Manabu Kanyama, Hajime Minakuchi, Shigehiko Shinkawa, Hiroya Takiuchi, Kumiko Nawachi, Kenji Maekawa, Hikaru Arakawa, Takuo Fujisawa, Wataru Sonoyama, Atsushi Mine, Emilio Satoshi Hara, Takeshi Kikutani, Takuo Kuboki

Journal of dental education 76 ( 12 ) 1580 - 8 2012.12

Language：English Publishing type：Research paper (scientific journal) Publisher：12

This educational trial was an eight-day problem-based learning (PBL) course for fourth-year predoctoral students at Okayama University's dental school who interviewed elderly residents living in a nursing home. The purpose of this PBL course was to introduce geriatric dentistry to the students by allowing them, independently, to discover the clinical problems of elderly patients as well as the solutions. The sixty-five students were divided into nine small groups and received patient information (age, gender, degree of care needed, medical history, food type, medications, and oral condition) in datasheets before visiting the nursing home. Each group of students directly interviewed one patient and the caregivers and identified the patient's medical, psychological, and social problems. After the interview, the students participated in a PBL tutorial to delineate a management approach for the patient's problems. To measure the efficacy of this program, the students completed a questionnaire before and after the course regarding their level of understanding of and attitudes toward geriatric dentistry, clinical research, and self-study. The results showed that student's perceptions of their knowledge about and attitudes toward oral health care for the elderly significantly increased after the PBL course, which suggests that such tutorials should be an option for dental curricula.

Multiple sleep bruxism data collected using a self-contained EMG detector/analyzer system in asymptomatic healthy subjects. Reviewed International journal

Hajime Minakuchi, Chiyomi Sakaguchi, Emilio S Hara, Kenji Maekawa, Yoshizo Matsuka, Glenn T Clark, Takuo Kuboki

Sleep & breathing = Schlaf & Atmung 16 ( 4 ) 1069 - 72 2012.12

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

PURPOSE: Small, self-contained electromyographic (EMG) detector/analyzer (D/A) devices have become available for the detection of jaw muscle activity events above threshold. These devices claim to be less intrusive to the subjects sleep so it is less prone to induce disturbed sleep. The objective of this study was to evaluate for night-to-night variability and examine for a systematic alteration on the first night in EMG levels. METHODS: Ten asymptomatic healthy volunteers (mean age, 26.8 ± 3.78) were recorded for six sequential nights in their home environment using EMG D/A system. The device yields a nightly EMG level above threshold score on a 0-4 level. Because the data are categorical and nonparametric, the data of the ten subjects across six nights were submitted to a Friedman repeated measures ANOVA. The significant level was set as alpha equal to 0.05. RESULTS: The median and mode values of the subjects were tabulated and analyzed and we did not find a significant difference in EMG D/A level across the six nights (p = 0.287, Kendall's coefficient of concordance = 0.124, Friedman two-way repeated measures ANOVA). The data did show clear and substantial night-to-night variability. CONCLUSION: Substantial night-to-night variability in masseter EMG activity levels was clearly observed in our subjects. There was no evidence of a suppressed or elevated first-night effect-like variability on masseter muscle EMG level seen in these subjects using a small portable self-contained EMG detector/analyzer. These data suggest that recordings should be at least 5-6-nights duration to establish a reasonable measure of an individual's average nightly masseter EMG level.

DOI： 10.1007/s11325-011-0602-1

Multiple sleep bruxism data collected using a self-contained EMG detector/analyzer system in asymptomatic healthy subjects Reviewed International journal

Hajime Minakuchi, Chiyomi Sakaguchi, Emilio S. Hara, Kenji Maekawa, Yoshizo Matsuka, Glenn T. Clark, Takuo Kuboki

SLEEP AND BREATHING 16 ( 4 ) 1069 - 1072 2012.12

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

Small, self-contained electromyographic (EMG) detector/analyzer (D/A) devices have become available for the detection of jaw muscle activity events above threshold. These devices claim to be less intrusive to the subjects sleep so it is less prone to induce disturbed sleep. The objective of this study was to evaluate for night-to-night variability and examine for a systematic alteration on the first night in EMG levels.
Ten asymptomatic healthy volunteers (mean age, 26.8 +/- 3.78) were recorded for six sequential nights in their home environment using EMG D/A system. The device yields a nightly EMG level above threshold score on a 0-4 level. Because the data are categorical and nonparametric, the data of the ten subjects across six nights were submitted to a Friedman repeated measures ANOVA. The significant level was set as alpha equal to 0.05.
The median and mode values of the subjects were tabulated and analyzed and we did not find a significant difference in EMG D/A level across the six nights (p = 0.287, Kendall's coefficient of concordance = 0.124, Friedman two-way repeated measures ANOVA). The data did show clear and substantial night-to-night variability.
Substantial night-to-night variability in masseter EMG activity levels was clearly observed in our subjects. There was no evidence of a suppressed or elevated first-night effect-like variability on masseter muscle EMG level seen in these subjects using a small portable self-contained EMG detector/analyzer. These data suggest that recordings should be at least 5-6-nights duration to establish a reasonable measure of an individual's average nightly masseter EMG level.

DOI： 10.1007/s11325-011-0602-1

CCN2非依存性モジュールの軟骨再生能力(Cartilage regeneration potential of CCN2 independent modules)

El Kader Tarek Abd, Kubota Satoshi, Nishida Takashi, Hattori Takako, Aoyama Eriko, Janune Danilo, Kuboki Takuo, Takigawa Masaharu

日本再生歯科医学会誌 10 ( 1 ) 50 - 50 2012.12

A prosthodontic medicine with collaboration between medical and dental sciences. Reviewed

Takuo Kuboki

Journal of prosthodontic research 56 ( 4 ) 227 - 8 2012.10

Matrix metalloproteinase-8 is the major potential collagenase in active peri-implantitis. Reviewed

Hikaru Arakawa, Junji Uehara, Emilio Satoshi Hara, Wataru Sonoyama, Aya Kimura, Manabu Kanyama, Yoshizo Matsuka, Takuo Kuboki

Journal of prosthodontic research 56 ( 4 ) 249 - 55 2012.10

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

PURPOSE: Current clinical procedures to control or regenerate bone loss due to peri-implantitis are not predictable neither accomplish complete resolution. Therefore, early detection of the onset and the active periods of bone loss are crucial for prevention of extensive peri-implant bone resorption. This study aimed to determine a possible association between the presence of collagenases (MMP-1, MMP-8 and MMP-13) in peri-implant sulcular fluid (PISF) and active periods of bone loss by annually adjusted vertical bone loss (AVBL) measurements. METHODS: Intended sample consisted of 76 consecutive patients who received oral implant treatment at the Fixed Prosthodontic Clinic of Okayama University Hospital from 1990 to 2000. Twelve subjects were lost to follow-up or refused to participate. Consequently, the actual sample consisted of 64 patients who were followed-up for at least one year. Those patients with AVBL>0.6mm were included in the severe peri-implantitis group, and randomly selected, age-, gender- and implantation site-matched healthy patients (AVBL<0.3mm) comprised the control group. PISF samples were collected from both groups and further analyzed by western blot for detection of collagenases. RESULTS: Four patients presented severe peri-implantitis. MMP-8 was the only collagenase detected in peri-implant sites with ongoing bone loss. PISF samples from control group showed no positive reactions to any collagenase. CONCLUSION: This study showed MMP-8 as a possible marker for progressive bone loss in peri-implantitis.

DOI： 10.1016/j.jpor.2012.07.002

Matrix metalloproteinase-8 is the major potential collagenase in active peri-implantitis Reviewed

Hikaru Arakawa, Junji Uehara, Emilio Satoshi Hara, Wataru Sonoyama, Aya Kimura, Manabu Kanyama, Yoshizo Matsuka, Takuo Kuboki

JOURNAL OF PROSTHODONTIC RESEARCH 56 ( 4 ) 249 - 255 2012.10

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

Purpose: Current clinical procedures to control or regenerate bone loss due to peri-implantitis are not predictable neither accomplish complete resolution. Therefore, early detection of the onset and the active periods of bone loss are crucial for prevention of extensive pen-implant bone resorption. This study aimed to determine a possible association between the presence of collagenases (MMP-1, MMP-8 and MMP-13) in peri-implant sulcular fluid (PISF) and active periods of bone loss by annually adjusted vertical bone loss (AVBL) measurements.
Methods: Intended sample consisted of 76 consecutive patients who received oral implant treatment at the Fixed Prosthodontic Clinic of Okayama University Hospital from 1990 to 2000. Twelve subjects were lost to follow-up or refused to participate. Consequently, the actual sample consisted of 64 patients who were followed-up for at least one year. Those patients with AVBL > 0.6 mm were included in the severe peri-implantitis group, and randomly selected, age-, gender- and implantation site-matched healthy patients (AVBL <0.3 mm) comprised the control group. PISF samples were collected from both groups and further analyzed by western blot for detection of collagenases.
Results: Four patients presented severe peri-implantitis. MMP-8 was the only collagenase detected in pen-implant sites with ongoing bone loss. PISF samples from control group showed no positive reactions to any collagenase.
Conclusion: This study showed MMP-8 as a possible marker for progressive bone loss in peri-implantitis. (C) 2012 Japan Prosthodontic Society. Published by Elsevier Ireland. All rights reserved.

DOI： 10.1016/j.jpor.2012.07.002

矯正歯科治療患者の口腔関連Quality of Lifeの評価

歯科矯正治療を受診する患者の口腔関連Quality of Lifeの評価

古森紘基, 川邉紀章, 片岡伴記, 白賀のり子, 大野彩, 窪木拓男, 山城隆

日本矯正歯科学会大会プログラム・抄録集 71回 198 - 198 2012.9

古森紘基, 川邉紀章, 片岡伴記, 白賀のり子, 大野彩, 窪木拓男, 山城隆

中・四国矯正歯科学会雑誌 24 ( 1 ) 106 - 106 2012.8

Identification of novel transcription-regulating genes expressed during murine molar development. Reviewed International journal

Kenta Uchibe, Hirohito Shimizu, Shigetoshi Yokoyama, Takuo Kuboki, Hiroshi Asahara

Developmental dynamics : an official publication of the American Association of Anatomists 241 ( 7 ) 1217 - 26 2012.7

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

BACKGROUND: The mechanism of tooth development is a complex process regulated by numerous genes including transcription factors, growth factors, and other intra- and extracellular molecules. Especially, transcription factors play a central role in gene expression, regulating a wide spectrum of biological processes including organogenesis. Substantial evidence has been demonstrated by a number of studies using genetically engineered animal models. However, detailed molecular mechanisms of tooth development have not been completely elucidated, partially because numerous genes that play essential roles in tooth development remain unidentified. RESULTS: In this study, we conducted an expression-based screening using gene expression database and in situ hybridization assays. Based on the gene expression database "EMBRYS," 207 out of 1,520 genes were expressed in the maxillary and/or mandibular processes and thus were selected for further analysis by section in situ hybridization. Among these candidates, 28 genes were newly identified as potential factors associated with tooth development by in situ hybridization assays with frontal sections of embryonic day 13.5 and 14.5 mouse embryos. The expression patterns were also examined at embryonic day 16.5 and 18.5. CONCLUSIONS: These results will contribute to elucidating the mechanisms of tooth development and to improving the technology for regeneration of tooth.

DOI： 10.1002/dvdy.23808

軟骨細胞と骨芽細胞に対するCCN2独立モジュールの影響(Effects of CCN2 independent modules on chondrocytic and osteoblastic cells)

El Kader Tarek Abd, Kubota Satoshi, Nishida Takashi, Hattori Takako, Aoyama Eriko, Janune Danilo, Kuboki Takuo, Takigawa Masaharu

日本骨代謝学会学術集会プログラム抄録集 30回 232 - 232 2012.7

Bonding effectiveness of a new 'multi-mode' adhesive to enamel and dentine. Reviewed International journal

Masao Hanabusa, Atsushi Mine, Takuo Kuboki, Yasuko Momoi, Annelies Van Ende, Bart Van Meerbeek, Jan De Munck

Journal of dentistry 40 ( 6 ) 475 - 84 2012.6

Language：English Publishing type：Research paper (scientific journal) Publisher：6

OBJECTIVES: Self-etch adhesives are well adopted in general practice, obviously primarily thanks to their ease of use and fast application time. Nevertheless, phosphoric acid is still often recommended to beforehand etch enamel following a so-called 'selective' enamel-etch technique, this in particular when most cavity margins end in enamel. The purpose of this study was to test if a new one-step adhesive can be applied in a multi-mode manner, this following different, either 'full' or 'selective', self-etch and etch-and-rinse approaches. Specific research hypotheses tested were that prior phosphoric-acid etching did not affect the bonding effectiveness of the one-step adhesive to enamel and dentine, and that the bonding effectiveness to dentine was also not affected when the adhesive was applied either following a 'dry-bonding' or 'wet-bonding' etch-and-rinse technique. METHODS: The micro-tensile bond strength (μTBS) of the one-step self-etch adhesive G-Bond Plus (GC, Tokyo, Japan; 1-SEA) was measured when it was bonded to bur-cut enamel following either a 'self-etch' or an 'etch-and-rinse' adhesive protocol, and to bur-cut dentine when applied following either a 'self-etch', a 'dry-bonding' or a 'wet-bonding' etch-and-rinse adhesive protocol. Bond-strength testing was corroborated by ultra-structural analysis of the interfacial interaction at enamel and dentine using transmission electron microscopy (TEM). RESULTS: Prior phosphoric-acid etching significantly increased the bonding effectiveness of the 1-SEA to enamel. A clearly enhanced micro-retentive surface was revealed by TEM. To dentine, no statistically significant difference in bonding effectiveness was recorded when the 1-SEA was either applied following a self-etch or both etch-and-rinse approaches. The 'dry-bonding' etch-and-rinse protocol was significantly more effective than its 'wet-bonding' version. TEM however revealed indications of low-quality hybridisation following both etch-and-rinse approaches, in particular in the form of a porous and poorly resin-infiltrated collagen mesh. CONCLUSIONS: While phosphoric-acid etching definitely improved bonding of the one-step self-etch adhesive to enamel, one should be more careful with additional phosphoric-acid etching of dentine. Although the bond strength was not reduced, the resultant adhesive interface appeared ultra-structurally more vulnerable to biodegradation.

DOI： 10.1016/j.jdent.2012.02.012

Videoendoscopic assessment of swallowing function to predict the future incidence of pneumonia of the elderly Reviewed

N. Takahashi, T. Kikutani, F. Tamura, M. Groher, T. Kuboki

Journal of Oral Rehabilitation 39 ( 6 ) 429 - 437 2012.6

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

The purpose of the present study was to examine what dysphagic signs identified by videoendoscopy (VE) could predict the incidence of pneumonia and body weight loss in elderly patients living in nursing homes. This study was performed at six nursing care facilities in Japan from March 2007 to February 2009. The 148 subjects (85·1±8·0years, male/female: 43/105) were evaluated for their feeding and swallowing movements by clinical and VE examinations during the consumption of a regular meal. The VE examination items included the existence/absence of pharyngeal residue, laryngeal penetration, and aspiration of food and saliva. The patients were followed-up for 3months with individualized feeding therapy based on the results of the clinical/VE examination at baseline, and the incidence of pneumonia was examined as the primary outcome. In patients without pneumonia, the body weight change was also measured as a secondary outcome. The risk factors for pneumonia and body weight loss (of 3% or more) were identified among the clinical/VE examination items by a Cox proportional hazard analysis. Even with elaborative feeding therapy, 12 (8·1%) of the 148 patients developed pneumonia during the 3months follow-up period. The existence of signs of 'silent aspiration of saliva' or 'aspiration of saliva' detected by VE examination was a significant risk factor for both pneumonia and a body weight loss of 3% or more. This study shows that 'aspiration of saliva' detected by VE is a significant risk factor for both pneumonia and body weight loss in elderly patients living in nursing homes. © 2012 Blackwell Publishing Ltd.

DOI： 10.1111/j.1365-2842.2011.02286.x

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