An experiment study of osteogenesis of Ad-VEGF165 transfected human bone marrow mesenchymal stem cells in vitro

ObjectiveTo evaluate the effect of osteogenic potential on human marrow mesenchymal stem cells(hMSCs) transferred with human vascular endothelial growth factor(VEGF) gene by adenovirus.MethodshMSCs were isolated from human marrow, cultured in vitro and randomly divided into 3 groups:Ad-VEGF165 group: adding 1 × 1 010OPU/ml Ad-VEGF in hMSCs culture fluid after incubating 24 hours, changing into ordinary complete culture and continuing culturing; Positive control group: Cultured hMSCs with 1 nmol/L dexamethasone,10 mmol/L glycerophosphate and 50 mg/L vitamin C, exchanging this conditioned medium twice a week; blank control group:no special treatment but culturing hMSCs in DMEM. To evaluate osteogenesis competence, Von Kossa±s staining and a quantitative alkaline phosphates(ALP) activity analysis were performed after 2 weeks treatment.ResultsThe calcified nodes formed after 2 weeks treatment in Ad-VEGF165 group and Positive control group but not in blank control group. ALP activities in Ad-VEGF165 group, Positive control group and blank control group were(7.91 ± 0.90)u/L,(8.18 ± 0.76 u/L) and (3.46 ± 0.49)u/L respectively. The differences were no statistical significance between Ad-VEGF165 group and positive control group(P >0.05), but Ad-VEGF165 group and Positive control group were significantly different with blank control group(P <0.05).ConclusionAdenovirus mediated VEGF165 gene can transfect hMSCs and promote osteogenesis of hMSCs.