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[Effects of secretive bone morphogenetic protein 2 induced by gene transfection on the biological changes of NIH3T3
Wei-bin Sun1, Juan Wang, Chun Lu
1Department of Periodontology, Stomatological College, Nanjing Medical University, Nanjing 210029, China. wbsun@njmu.edu.cn
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 cells incubated with secretive BMP2 that was induced by gene transfection through transwell.
Methods:
Eukaryonic expression vector (pcDNA3.1-B2) was transfered into NIH3T3 cells with Sofast, 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. NIH3T3 cells were co-cultured with hBMP2 gene transfecting cells 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 cells. 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 cells.
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