| Project/Area Number |
12470419
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
補綴理工系歯学
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| Research Institution | OKAYAMA UNIVERSITY |
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Principal Investigator |
KANYAMA Manabu Okayama Univ, Dental Hospital, Lecture, 歯学部・附属病院, 講師 (90294420)
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| Co-Investigator(Kenkyū-buntansha) |
NAKANISHI Tohru Okayama Univ, Graduate Sch of Med and Dent, Associate Professor, 大学院・医歯学総合研究科, 助教授 (30243463)
TAKIGAWA Masaharu Okayama Univ, Graduate Sch of Med and Dent, Professor, 大学院・医歯学総合研究科, 教授 (20112063)
KUBOKI Takuo Okayama Univ, Graduate Sch of Med and Dent, Associate Professor, 大学院・医歯学総合研究科, 助教授 (00225195)
ARAKAWA Hikaru Okayama Univ, Graduate Sch of Med and Dent, Research Associate, 大学院・医歯学総合研究科, 助手 (30304314)
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| Project Period (FY) |
2000 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥8,200,000 (Direct Cost: ¥8,200,000)
Fiscal Year 2001: ¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 2000: ¥5,900,000 (Direct Cost: ¥5,900,000)
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| Keywords | Adenovirus vector / CTGF / MC3T3-E1 cell / Tooth extraction socket / Titanium / Ex vivo gene delivery / MC3T3-El cell / 抜歯創時癒過程 / BMP遺伝子 / CTGF遺伝子 / MC3T3-E1細胞 / TGF-β遺伝子 |
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| Research Abstract |
The purpose of this study was to contribute to shortening the healing period of the dental implant and expanding its application by ex vivo gene delivery using an adenovirus vector. It has been well-documented that connective tissue growth factor (CTGF) is up-regulated in the healing process at the bone fracture site in vivo and known as a potent stimulator for the proliferation and differentiation of osteoblasts in vitro. Therefore, CTGF gene transfer to the osteoblasts could be a promising strategy to accelerate bone formation. First, we investigated the possibility of recombinant adenovirus to transfer CTGF genes to a mouse osteoblastic (MC3T3-E1) cell line in vitro. Recombinant adenovirus encoding the LacZ gene and the human CTGF gene with CAG promotor (Ax1CACTGF) were applied to the MC3T3-E1 cells with 5, 10 and 50 multiplicity of infection (MOI). As the result, the MOI 50 transfection dosage produced almost 80% X-gal staining of the cells without any obvious cell damages and Ax1CACTGF transfection caused a marked up-regulation in CTGF mRNA expression and CTGF protein even 7 days after transfection. Second, we investigated the spatial and temporal expression of CTGF in the rat tooth extraction sockets to know the mechanisms of new alveolar bone formation. As the result, CTGF was expressed in the endothelial cells migrating into the granulation tissue at the sockets during 4 days after tooth extraction. Osteoblast-like cells proliferated in the sockets with CTGF expression at 4, 7, 10 and 14 days after extraction. Finally, we tried the ex vivo gene delivery, but we could not isolate osteoblasts from the defects of rat alveolar bone.
Based on these findings, adenovirus-mediated CTGF gene transduction to the cultured osteoblast-like cells was highly successful. However, in order to do the ex vivo gene transfer, it is necessary to develop the new methods of isolating osteoblasts.
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