Effects Of Calcitonin Gene-related Peptide On Osteogenic Differentiation And Angiogenesis Of Human Periodontal Ligament Stem Cells In Vitro

Objective To investigate the effects of calcitonin gene-related peptide?CGRP?on osteogenic differentiation and angiogenesis of human periodontal ligament stem cells?hPDLSCs?in vitro.Methods The hPDLSCs were isolated and cultured by tissue block method,identification of cells by flow cytometry and immunohistochemistry.Different concentrations of CGRP were given.CCK-8 method was used to analyze the effect of CGRP on the proliferation of hPDLSCs.The mRNA expressions of alkaline phosphatase?ALP?,core binding factor 2?RUNX2?,osteocalcin?OCN?and vascular endothelial growth factor?VEGF?were detected by Raeltime-PCR.The expression of Runx2,OCN and VEGF protein was detected by Western blot.ALP staining was performed 7 days after osteogenic induction,and alizarin red staining was performed 21days after osteogenic induction.Results.Cells isolated and cultured from human periodontal ligament tissue were identified to be consistent with the characteristics of mesenchymal stem cells.CCK-8results showed that CGRP promoted the proliferation of hPDLSCs within a certain range?P<0.05?.The results of RT-PCR and Western blot showed that 10^-8 mol/L CGRP significantly up-regulated the expression of ALP,RUNX2,OCN and VEGF gene and protein?P<0.05?.ALP and alizarin red staining results showed that 10^-8 mol/L CGRP significantly promoted the expression of ALP and the formation of calcium nodules in osteogenic differentiation of hPDLSCs.Conclusion 10^-8 mol/L CGRP can promote osteogenic differentiation and angiogenesis of hPDLSCs in vitro.