| Project/Area Number |
10671373
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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 |
Orthopaedic surgery
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| Research Institution | Kyoto Prefectural University of Medicine |
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Principal Investigator |
NAMIE Kazuo Kyoto Prefectural University of Medicine, the medical department, Assistant, 医学部, 助手 (10295652)
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| Co-Investigator(Kenkyū-buntansha) |
KUBO Toshikazu Kyoto Prefectural University of Medicine, the medical department, Associate Professor, 医学部, 助教授 (20178031)
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| Project Period (FY) |
1998 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥2,900,000 (Direct Cost: ¥2,900,000)
Fiscal Year 1999: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1998: ¥1,700,000 (Direct Cost: ¥1,700,000)
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| Keywords | gene therapy / adenovirus vector / cartilage repair / cartilage injury / ex vivo / drug delivery system / graft / chondrocyte |
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| Research Abstract |
Cartilage defects do not recover and the damaged cartilage will usually develop degeneration in the long term, because ordinarily articular cartilage does not regenerate. To date, various experimental studies such as autologous and allograft chondrocyte transplantation have been conducted to achieve better repair of the defects. As a means to activate metabolic activity of grafted chondrocytes, cytokines could be useful. Another approach, gene therapy, has attracted attention in a wide range of specialties, and its application to joint diseases has also been studied. We investigated the usefulness of an ex vivo gene delivery method using an adenovirus in chondrocyte grafting for cartilage defects
β-galactosidase gene (LacZ) was transfected using an adenovirus vector to chondrocytes isolated from rat joints. The cells were then embedded into collagen gel, and LacZ expression in the gel was examined using 5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside (X-gal) staining ; β-galactosidase activity was also measured. The collagen gel containing transfected chondrocytes was grafted to the experimental cartilage defects, and the expression of delivered gene was histologically examined after X-gal staining of the tissue containing the grafted area.
X-gal-positive chondrocytes in the gel accounted for 82% at one week and 55% at 8 weeks after gene delivery.β-galactosidase activity decreased with time, but its expression was maintained even at 8 weeks after gene delivery. Chondrocytes used in the allograft maintained their morphology, and the expression of delivered gene continued during the 8-week period.
We consider that ex vivo gene delivery using an adenovirus vector would be a promising technique for cartilage repair because this method can realize effective action of cytokines and other remedies specifically on the damaged area.
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