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

Objective:To evaluate the effect of osteogenic potential on human marrow mesenchymal stem cells (hMSCs) transferred with human vascular endothelial growth factor(VEGF) gene by adenovirus. Methods :hMSCs were isolated from human marrow, cultured in vitro and randomly divided into 3 groups:Ad-VEGF165 group: adding 1×1010OPU/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. Results :The 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). Conclusion:Adenovirus mediated VEGF165 gene can transfect hMSCs and promote osteogenesis of hMSCs.

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

Objective:To evaluate the effect of osteogenic potential on human marrow mesenchymal stem cells (hMSCs) transferred with human vascular endothelial growth factor(VEGF) gene by adenovirus. Methods :hMSCs were isolated from human marrow, cultured in vitro and randomly divided into 3 groups:Ad-VEGF165 group: adding 1×1010OPU/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. Results :The 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). Conclusion:Adenovirus mediated VEGF165 gene can transfect hMSCs and promote osteogenesis of hMSCs.