| Project/Area Number |
14571365
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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 | KANAZAWA UNIVERSITY |
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Principal Investigator |
TSUCHIYA Hiroyuki Graduate School of Medicine Kanazawa Univ., Associated Prof., 大学院医学系研究科, 助教授 (40227434)
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| Co-Investigator(Kenkyū-buntansha) |
SAKURAKICHI Keisuke raduate School of Medicine Kanazawa Univ., Assistant Prof., 大学院医学系研究科, 助手 (60334752)
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| Project Period (FY) |
2002 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2003: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2002: ¥2,100,000 (Direct Cost: ¥2,100,000)
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| Keywords | normal mesenchymal cell / recombinant human bone morphogenetic protein-2 / bone formation / gene transfection / distraction osteogenesis / bone defect / 骨形成因子 / BMP-2 / 仮骨延長 / 遺伝子治療 / 創外固定 |
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| Research Abstract |
We made the plasmid vector coding fusion protein of green fluorescein protein and recombinant human bone morphogenetic protein-2(rhBMP-2).The plasmid vector were transfected into C2C12, and NIH3T3, fibroblast cell line of mouse with lipofection.Expression of mRNA of rhBMP-2 were increased.Transfected cells showed GFP expression with laser scanning microscopy, but no relation between expression of rhBMP-2 and GFP with immunohistological study.C2C12 cells transfected with GFP-rhBMP-2 shows increasing of ALP activity.
Type I collagen and transfectd bone marrow stromal cells harvested from rat's femur, were transplantation into rat biceps femorins and 5mmfemoral bone defect made with external fixators.Rhoentogenographic study showed no bone formation.After plantation of transfected cells and collagen, lengthening of femur were done 0.5mm per day for 10days.The results were the same.
Gene coding GFP-rhBMP-2 were transfected into normal mesenchymal cells, but did not show increaeing of production of fusion protein with immunohistochemically.Transfected cellsshowed slight increase of ALP activity under in vivocircumference, but dose not showed heterotopic bone formation or acceleration of bone formation.
It needed to change the construction of GFP-rhBM-2 gene, or to swich the vector from plasmid to virus.
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