Acceleration of distraction osteogenesis with recombinant human bone morphogenetic protein-2 transfected into normal mesenchymal cell.

重组人骨形态发生蛋白2转染正常间充质细胞加速牵引成骨。

• 批准号: 14571365
• 负责人: TSUCHIYA Hiroyuki
• 金额: $ 2.24万
• 依托单位: KANAZAWA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14571365/
• 关键词: 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

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.

我们制作了编码绿色荧光素蛋白与重组人骨形态发生蛋白2（rhBMP-2）融合蛋白的质粒载体，用脂质体转染小鼠C2C12、NIH3T3等成纤维细胞系，rhBMP-2 mRNA表达增加。转染细胞激光扫描显微镜下可见GFP表达， 免疫组织学研究显示rhBMP-2和GFP的表达没有相关性。转染GFP-rhBMP-2的C2C12细胞显示ALP活性增加。从大鼠股骨收集I型胶原和转染的骨髓基质细胞，移植到大鼠股二头肌和外固定器制作的5mm股骨缺损处。Rhoentogenography研究显示没有 转染细胞和胶原植入后，每天延长股骨0.5mm，持续10天，结果相同。将编码GFP-rhBMP-2的基因转染到正常间充质细胞中，但免疫组化未显示融合蛋白产量增加。转染细胞在体内周情况下显示ALP活性略有增加，但不增加。 表现出异位骨形成或骨形成加速。需要改变GFP-rhBM-2基因的构建，或将载体从质粒转为病毒。