Experimental Research Of ARC Enhances Osteogenesis Differentiation Of Human Osteoblast And Prevents Its Apoptosis

Objective:To investigate the effect of ARC on apoptosis and osteogenesis differentiation of human osteoblast under Zoledronate treatment.Materials and methods:(1)Lentivirus vector mediated ARC overexpressing was constructed and then transduct human osteoblast and the transduction efficiency was confirmed by western blot and RT-PCR.(2)MTT method was used to detect the proliferation ability of OB-CON and OB-NOL3 under the treatment of different concentrations(0μM,1μM,5μM)of Zoledronate.(3)Flow cytometry method was applied to identity cell apoptosis of OB-CON and OB-NOL3 under the treatment of different concentrations of Zoledronate.(4)ALP staining and quantitative analysis were performed to detect the osteogenesis differentiation of human osteoblast at day 7 and day 14.(5)Real-time PCR and western blot analysis were used to detect the osteogenesis differentiation of OB-CON and OB-NOL3.(6)Ectopic osteogenesis in nude mice was further performed to detect the osteogenesis differentiation of ARC overexpressed human osteoblast.Results:(1)ARC overexpressed human osteoblast showed higher proliferation ability than control group under different concentrations of Zoledronate treatment.(2)After three days of Zoledronate treatment,the percentage of cell apoptosis of OB-CON and OB-NOL3 were 3.87% and 2.83% in 0μM group,13.35% and 8.15% in 1μM group,26.96% and 17.4% in 5μM group.(3)ALP staining and quantitative analysis revealed ARC overexpressed human osteoblast showed higher osteogenesis differentiation ability than control groups.(4)Ectopic osteogenesis in nude mice further confirmed higher osteogenesis differentiation ability in ARC overexpressed group.Conclusions: ARC can reverse the inhibitory effect of Zoldronate on osteoblasts.ARC can also promote osteogenesis differentiation of osteoblasts and inhibit their apoptosis.