| Project/Area Number |
16591948
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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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 |
SUZUKI Koji Okamaya University Hospital, Research Associate, 医学部・歯学部附属病院, 助手 (30304322)
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| Co-Investigator(Kenkyū-buntansha) |
KUBOKI Takuo Okayama University Graduate School of Medicine, Dentistry, Pharmaceutical Sciences, Professor, 大学院医歯薬学総合研究科, 教授 (00225195)
KANYAMA Manabu Okamaya University Hospital, Lecturer, 医学部・歯学部附属病院, 講師 (90294420)
峯 篤史 岡山大学, 医学部・歯学部附属病院, 助手 (60379758)
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| Project Period (FY) |
2004 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2006: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2005: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2004: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Keywords | CTGF / adenovirus / 遺伝子導入 / 口腔インプラント / ベクター / マウス骨芽細胞株MC3T3-E1 cell / 結合組織成長因子 |
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| Research Abstract |
CTGF gene transfer to the osteoblasts could be a promising strategy to accelerate bone formation. In this study, we investigated the possibility of recombinant adenovirus to transfer CTGF genes to a mouse osteoblastic (MC3T3-E1) cell line in vitro, and bone regeneration process after tooth extraction.
Rrecombinant adenovirus encoding the LacZ gene (Ax1CALacZ) were applied to the MC3T3-E1 cells with 5, 10 and 50 multiplicity of infection (MOI). At one day after transfection, those cells were stained with X-gal. As the result, the MOI 50 transfection dosage produced almost 80% X-gal staining of the cells without any obvious cell damages. Recombinant adenovirus encoding the human CTGF gene with CAG promotor (Ax1CACTGF) were fabricated and applied to the MC3T3-E1 cells in the MOI 50 dose. CTGF mRNA translation and protein production by the transfected cells were assessed by RT-PCR and western blotting of the harvested cells using a prepared primer set and an anti-human-CTGF antibody at 1, 3 and 7 days after transfection. Ax1CACTGF transfection caused a marked up-regulation in CTGF mRNA expression and CTGF protein even 7 days after transfection.
The sequential phase appearances of extraction wound healing were clearly observed by histological examination from 2 to 14 days after extraction. At 7 days after extraction, infilling with woven bone was observed from the socket margin with trabecular bone radiating toward the center of the socket, and at 14 days after, the extraction socket was almost fully filled by newly generated bone that underwent remodeling During any healing processes observed in the extraction wound, no cartilage-like cells were evident in the socket as determined by S-0 staining
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