人骨形成蛋白7重组腺病毒载体的构建及其在牙周膜细胞中的表达

Objective To investigate the expression of adenovirus-transfected human bone morphogenetic protein-7 (BMP-7) in human periodontal ligament cells(PDLCs) and its effect on PDLCs proliferation after transfection. Methods The BMP-7 fragment was amplified by PCR according to the pCMV-SPORT6-BMP-7, and the fragment was cloned into pShuttle-CMV-BMP-7, then it was homogenously recombined with pBHGE3 in 293 cells to obtain adenovirus expression vector containing BMP-7 (Ad-BMP-7). Ad-BMP-7 was identified and the titer of virus was measured after amplification and purification. Ad-RMP-7 transfected human PDLCs in vitro. The BMP-7 protein expression and proliferation of transfected PDLCs were detected by Western blot and MTT assay respectively. Results PCR analysis confirmed that the human BMP-7 gene was successfully inserted into the adenovirus vector. The titer of the recombinant adenovirus was 1.785×1012 pfu/ml. Expression of BMP-7 protein was detected in PDLCs transfected with Ad-BMP-7. The MTT test showed no significant difference between PDLCs transfected with or without Ad-BMP-7. Conclusions Adenovirus expression vector containing BMP-7 can transfect human PDLCs successfully with high expresion of BMP-7 in PDLCs in vitro.

Construction of recombinant adenoviruse vector containing bone human morphogenetic protein-7 and its expression in human periodontal ligament cells