Li Xiang-dong, Hu Yun-yu
Institute of Orthopedics, Xijing Hospital, Fourth Military Medical University, Xi'an Shaanxi, P. R. China 710032.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2003 May;17(3):177-9.
To determine whether fibroblasts can be used to promote endochondral bone formation in vivo by transfer of human bone morphogenetic protein-2(hBMP-2) into fibroblasts.
pcDNA3-hBMP-2 was constructed by use of gene clone and recombined technique. NIH3T3 fibroblasts were transfected with pcDNA3-hBMP-2. The positive cell clones were selected with G418. In NIH3T3 fibroblasts transferred with pcDNA3-hBMP-2, the expression of hBMP-2 was determined by in situ hybridization and immunohistochemical analysis; alkaline phosphatase activity was measured. hBMP-2-producing fibroblasts were implanted into nude mouse muscle to observe endochondral bone formation in vivo.
pcDNA3-hBMP-2 was successfully constructed. In NIH3T3 fibroblasts transfected with pcDNA3-hBMP-2, the BMP-2 expression was stable; alkaline phophatase activity was much higher than that in non-transfected NIH3T3 cells. Endochondral bone formation in vivo was observed at the site of implantation 4 weeks later.
Fibroblasts transfected by hBMP-2 gene can be used to promote endochondral bone formation in vivo.
通过将人骨形态发生蛋白-2（hBMP-2）导入成纤维细胞，确定成纤维细胞是否可用于促进体内软骨内骨形成。
利用基因克隆和重组技术构建pcDNA3-hBMP-2。用pcDNA3-hBMP-2转染NIH3T3成纤维细胞。用G418筛选阳性细胞克隆。在转染了pcDNA3-hBMP-2的NIH3T3成纤维细胞中，通过原位杂交和免疫组织化学分析确定hBMP-2的表达；测量碱性磷酸酶活性。将产生hBMP-2的成纤维细胞植入裸鼠肌肉中，观察体内软骨内骨形成情况。
成功构建了pcDNA3-hBMP-2。在转染了pcDNA3-hBMP-2的NIH3T3成纤维细胞中，BMP-2表达稳定；碱性磷酸酶活性远高于未转染的NIH3T3细胞。4周后在植入部位观察到体内软骨内骨形成。
用hBMP-2基因转染的成纤维细胞可用于促进体内软骨内骨形成。
