摘要
Background Bone morphogenetic proteins （BMPs）, which belong to the transforming growth factor beta superfamily, are powerful regulators of cartilage and bone formation. This study investigated the biological changes of NIH3T3 ceils incubated with secretive BMP2 that was induced by gene transfection through transwell.Methods Eukaryonic expression vector （pcDNA3. 1-B2 ） was transfered into NIH333 cells with Sofast^TM, a positive compound transfection agent. The positive cell clones were selected with G418. The cytoplasmic and extracellular expressions of BMP2 were determined by immunohistochemical stain and enzyme-linked immunosorbent assay. NIH333 ceils were co-cultured with hBMP2 gene transfecting ceils through transwell, and the ultrastructure, alkaline phosphatase activity and the expression of osteocalcin （the marker of osteogenetic differentiation） changes were observed.Results There were cytoplasmic and extracellular expressions of BMP2 in transfecting NIH3T3 ceils. The ultrastructural changes, the high activity of alkaline phosphatase and the positive stain of osteocalcin suggested the osteogenetic differentiation tendency of NIH3T3 cells co-cultured with transfecting NIH3T3 cells.Conclusion Secretive BMP2 that is induced by gene transfection could promote the osteogenetic differentiation of fibroblast ceils.
Background Bone morphogenetic proteins （BMPs）, which belong to the transforming growth factor beta superfamily, are powerful regulators of cartilage and bone formation. This study investigated the biological changes of NIH3T3 ceils incubated with secretive BMP2 that was induced by gene transfection through transwell.Methods Eukaryonic expression vector （pcDNA3. 1-B2 ） was transfered into NIH333 cells with Sofast^TM, a positive compound transfection agent. The positive cell clones were selected with G418. The cytoplasmic and extracellular expressions of BMP2 were determined by immunohistochemical stain and enzyme-linked immunosorbent assay. NIH333 ceils were co-cultured with hBMP2 gene transfecting ceils through transwell, and the ultrastructure, alkaline phosphatase activity and the expression of osteocalcin （the marker of osteogenetic differentiation） changes were observed.Results There were cytoplasmic and extracellular expressions of BMP2 in transfecting NIH3T3 ceils. The ultrastructural changes, the high activity of alkaline phosphatase and the positive stain of osteocalcin suggested the osteogenetic differentiation tendency of NIH3T3 cells co-cultured with transfecting NIH3T3 cells.Conclusion Secretive BMP2 that is induced by gene transfection could promote the osteogenetic differentiation of fibroblast ceils.
基金
This work is supported by grants from 135 Medical EmphasisFoundation of Jiangsu Province (RC 2002018) and Natural Scienceand Technology Foundation of Nanjing Medical University(CX2003006).
