VEGF And BMP-7 Gene Therapy Enhanced Bone Regeneration

Object: To construct vectors which can express bioactive vascular endothelial genetic factor 165 and vascular endothelial genetic factor 121 stably in MC3T3-E1.Methods: Gene VEGF165 and VEGF121 were obtained by RT-PCR from HL-60 and inserted into eukaryotic expression plasmid pcDNA3.1(+).The newly-constructed plasmids containing right sequence aim gene were transfected into MC3T3-E1 by lipofectin. After the cells had been limitedly diluted twice and selected by G418, the expression of VEGF gene was tested by ELISA, Western-blot and immunofluorescence essay methods, and bioactivity proved by MTT.Results: DNA sequencing indicated the sequences of the obtained VEGF165 and VEGF 121 were identical to the reported ones in Genebank, and the newly-constructed plasmids were VEGF165/pcDNA3.1(+)and VEGF121/pcDNA3.1 (+). The results of ELISA , Western-blot and immunofluorescence showed that thetransfected cells could express VEGF165 and VEGF121. And MTT proved the proteins were bioactive.Conclusion: MC3T3-E1 after transfected could express high level bioactive VEGF165 and VEGF121 stably.