| Project/Area Number |
09470320
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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 |
Orthopaedic surgery
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| Research Institution | KYOTO PREFECTURAL UNIVERSITY OF MEDICINE |
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Principal Investigator |
KUBO Toshikazu Kyoto Prefectural University of Medicine, Medical School, Associate Professor, 医学部, 助教授 (20178031)
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| Co-Investigator(Kenkyū-buntansha) |
TAKIGAWA Masaharu Okayama University, Dental School, Professor, 歯学部, 教授 (20112063)
SAWADA Kouhei Kyoto Prefectural University of Medicine, Medical School, Assistant, 医学部, 助手 (40264767)
石田 敏博 京都府立医科大学, 医学部, 助手 (20295653)
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥9,000,000 (Direct Cost: ¥9,000,000)
Fiscal Year 1998: ¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 1997: ¥5,700,000 (Direct Cost: ¥5,700,000)
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| Keywords | gene therapy / adenovirus vector / guinea pig / in vivo / drug delivery system / TGF-betaI / joint / 変形性関節症 / アデノウイルスペクター / LacZ / 軟骨細胞 / プロモーター |
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| Research Abstract |
We evaluated the in vivo applicability of adenovirus-mediated gene delivery in order to consider the feasibility of gene therapy for human joint diseases.
We directly injected vectors harbouring beta-galactosidase (beta-gal) or transforming growth factor (TGF)-beta1 gene into the joints of Hartley guinea pigs. Expressions of transduced beta-gal genes were examined by 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside (X-gal) staining and reverse transcription- polymerase chain reaction (RT-PCR). The levels of TGF-beta1 which was delivered to the joint and then transferred to the joint fluid, were assessed by enzyme-linked immunosorbent assay (ELISA). LacZ expression was observed in almost the entire synovial tissues and in chondrocytes on the surface of degenerated cartilage. Un expected effects of the direct vector inj ection on the other organs were also examined, no expression of delivered gene was observed. Therefore, intraarticular direct injection would achieve gene delivery limited to the joint cavity, possibly eliminating unecessary systemic effects. TGF-beta1 levels in the joint fluid following the gene delivery had been significantly higher than the levels in the controls for 2 weeks.
Direct gene delivery into the joint cavity is realistic with the in vivo gene delivery method using adenovirus vector, and it would be clinically applicable.
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