In Vitro Study On Accelerating Vascularization Of Dentin Powder-Concentrated Growth Factor Complex

ObjectiveThe application of concentrated growth factor as a scaffold material in dental pulp angiogenesis can not obtain an exact prognosis.In order to obtain a more ideal scaffold material,dentin powder(Dentin Powder,DP)and concentrated growth factor(Concentrated Growth Factor,CGF)were compounded to study whether the composite can accelerate angiogenesis in vitro,so as to provide assistance for dental pulp regeneration.MethodsDentin powder was obtained from isolated teeth,which was treated by physical and chemical methods to meet the requirements of cell culture,and concentrated growth factors were obtained from peripheral blood.Human umbilical vein endothelial cells(HUVEC)were cultured routinely in vitro.Firstly,ELISA was used to detect the content and release pattern of VEGF in dentin and concentrated growth factor.According to the results of ELISA,the complete endothelial culture medium treated with DP-CGF was prepared,while the control medium was not treated.The proliferation ability of human umbilical vein endothelial cells was detected by CCK-8 method,and the 450 nm absorbance values of the two groups were obtained,and the differences between the two groups were obtained by statistical analysis.In the cell scratch test to detect the migration ability of human umbilical vein endothelial cells,DP-CGF was used to treat the endothelial medium without serum and epithelial growth factor,while the control group was not treated.The images were taken under inverted microscope at 0h and 6h,and the data were measured by image processing software.After statistical analysis,the difference between the groups was obtained.Finally,after human umbilical vein endothelial cells were treated with dentin powder concentrated growth factor,the expression of target genes VEGF-A,VEGFR2,P100α,p100β,Akt and p27 were detected by real-time quantitative PCR and statistically analyzed.ResultsBoth dentin and concentrated growth factor contained VEGF,and the ability of local continuous release was found in both dentin and concentrated growth factor.Compared with the control group,the ability of cell proliferation in DP-CGF group was significantly enhanced,and the difference was statistically significant,the ability of cell migration was enhanced in DP-CGF group,and the difference was statistically significant,VEGF-A,VEGFR2,p100α,p100β and Akt genes in DP-CGF group were significantly up-regulated(P<0.05),while cell cycle related gene p27 was significantly down-regulated(P<0.05).ConclusionsIn vitro,dentin powder and concentrated growth factor contain a certain amount of VEGF and can be released continuously under certain conditions,and the complex of dentin powder and concentrated growth factor can effectively promote the proliferation and migration of vascular endothelial cells and greatly accelerate the process of angiogenesis,which has positive significance for dental pulp regeneration.Dentin powder combined with concentrated growth factor may affect the cell cycle of HUVEC through VEGF-PI3K-p27 signal pathway,thus improve its proliferation and migration ability and accelerate the process of vascularization.