Effects Of Platelet Concentrated Growth Factors On Angiogenic Properties Of Dental Pulp Cells And Endothelial Cells

Dental pulp revascularization has been widely applied in the dental clinics in recent years,which is demonstrated to promote continuous root development of necrotic immature teeth.Concentrate growth factors(CGF),as the newest platelet concentrates product,contains abundant growth factors,and can be used to induce tissue regeneration.In addition,CGF is easily available from whole blood and eliminates the cell components in the whole blood so that the risk of infectious disease can be avoided.CGF has been clinically shown to accelerate tissue healing in many fields,for example bone reconstruction,angiogenesis and dental implant.However,the angiogenic effects of CGF on dental pulp revascularization was rarely discussed.Objective: In order to supply the experimental basement of clinical usage,this study aimed to explore the angiogenic effects of concentrate growth factors extracts(CGFe)on human umbilical vein endothelial cells(HUVECs)in vitro.Methods: DPCs(dental pulp cells)were isolated from extracted teeth using the enzymatic digestion combined with tissue cultivation method.CGF was prepared from whole blood of healthy volunteers and than CGFe was made.Five groups are designed depending on the cell culture medium: 2% CGFe,5% CGFe,10% CGFe,15% CGFe and control.The proliferation activity of DPCs and HUVECs was detected by a CCK8 assay.The VEGF,CXCR4 and PDGF mRNA expression of DPCs was examined by qRTPCR.The VEGF,CXCR4 protein level of DPCs were detected by western blot.The migration of HUVECs was detected by a transwell assay.In-vitro tube formation assay was conducted to evaluate the angiogenic capacity of HUVECs.Results: The CCK-8 assay showed that CGFe promoted DPCs and HUVECs proliferation in a concentration dependent manner when the concentration of CGFe is below 10%(P<0.05);the effects of 10%CGFe and 15%CGFe showed no statistical difference for DPCs(P>0.05),whereas the 15%CGFe showed proliferation inhibition effect to HUVECs at 36h(P<0.05).Thus,the 10%CGFe was chosen as optimum concentration and 15%CGFe group was not set in the other assays.The expression of VEGF,CXCR4 and PDGF mRNA and VEGF,CXCR4 protein of dental pulp cells were upregulated by CGFe in a concentration dependent manner(P<0.05).CGFe significantly promoted migration of HUVECs and formation of tube-like structures compared with control group,and the stimulative effect was in a concentration dependent manner((P<0.05).Conclusion: CGF promoted dental pulp cells proliferation and differentiation in vitro;CGF promoted HUVECs proliferation,migration and vascular structure formation.