Research Of Different Doses Of Epigallocatechin Gallate On Osteoblasts In Vitro

Objective:To determine the maximum effect concentration(maximum inhibitory concentration or promoting concentration),the effects of different doses of epigallocatechin gallate on osteoblasts in vitro is studied.Methods:Osteoblasts isolated from the cranium of newborn SD rats are cultured in vitro,and the third generation cells are devided into 6 groups with the presence of 0,1,10,20,30 and 40 μmol/L epigallocatechin gallate cultured in DMEM medium+10% fetal bovine serum.The viability of osteoblasts,mineral node formation were determined using CCK-8,alizarin red staining,von kossa staining,and the activity of alkaline phosphatas,bone morphogenetic protein 2,vascular endothelial growth factor,Runt-related transcription factor 2 were determined using AKP,Elisa,Western blot,respectively.Results:The viability of osteoblasts treated with epigallocatechin gallate at 10~20μmol/L is significantly higher than that of the 0μmol/L group,and the concentration of 30μmol/L and above showed inhibitory effect on rat osteoblasts.Mineral node formation can be clearly observed after 7 days or 14 days culture at different concentrations of EGCG,and the number of mineral modules are significantly increased in osteoblasts cultured with epigallocatechin gallate at 10~20μmol/L when compared with the untreated cells.Alkaline phosphatase activity in 10-20 μmol / L group is significantly higher than that in other groups.Bone morphogenetic protein 2(BMP-2)and vascular endothelial growth factor(VEGF)test all shows that the concentration of 10~20μmol/L has significant promotion to the osteoblasts secrete BMP-2 and VEGF,and the concentration of 30μmol/L and above showed inhibitory effect on secrete BMP-2 and VEGF.It is determined that the concentration of 10~20μmol/L has significant promotion to the osteoblasts secrete BMP-2 protein and Runx2 protein.Conclusion:EGCG at the concentration of 10 ~ 20μmol / L has significant promotion to the proliferation of osteoblasts,and the concentration of 30μmol/L and above showed inhibitory effect on osteoblasts cultured in vitro.