| Co-Investigator(Kenkyū-buntansha) |
TUCHIDA Hiroyuki Yamagata University, Orthopaedic Surgery, Assistant Professor, 医学部, 助教授 (40250922)
HASIMOTO Junichi Yamagata University, Orthopaedic Surgery, Lecturer, 医学部, 助手 (10359565)
川前 金幸 山形大学, 医学部, 教授 (70254026)
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| Budget Amount *help |
¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 2005: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2004: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Research Abstract |
Previously, we demonstrated osteogeic potential of Adenoviral BMP2 gene transferred mesenchymal stem cell, in rat bone defect model. However, adenoviral exhibit effective gene transfer, viral vector also has toxic effect to the cells and grafted tissues. To overcome these toxic effect, HVJ envelope vector has been established (HVJ ; Sendai virus). HVJ with robust fusion activity was inactivated, and exogenous DNA was incorporated into the viral envelope by detergent treatment and centrifugation. HVJ envelope vector introduce plasmid DNA into in vitro and in vivo, furthermore, proteins, synthetic oligonucleotide, and drugs. And no report is found regarding osteogenic potential of mesenchymal stem cell, BMP gene transferred using HVJ envelope vector. This study was undertaken to determine whether direct injection of HVJ envelope vector containing BMP4 gene can be used of bone regeneration. Five Brown-Norway rats were used in this study. Using this vector introduced BMP4, we delivered BMP4 gene into rat femoral muscle by direct percutaneous injection. After injection, soft-ray roentgenogram was obtained at 2 and 4 weeks. At 4 weeks, the rats were sacrificed for histological inspection. Roentgenogram and histological sections demonstrated no bone formation in samples of 2 and 4 weeks after injection. We speculate that administration rout for this vector could be incorrect, and response and number of mesenchymal stem cells was inadequate. In vivo gene therapy require adequate amount of target cells, therefore, the possibility of low amount of intra muscular stem cells was considered in our experiment. Further experiment regarding adequate way for this gene therapy was thought to be necessary.
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