Effects Of VEGFA On Proliferation And Neurogenic Differentiation Of Rat Dental Pulp Stem Cells

Background and purposeNerve damage and degenerative diseases often lead to disability and death.At present,the treatment methods for this kind of disease are limited and the prognosis of patients is poor,so it is urgent to find an efficient and accurate treatment method.Dental pulp stem cells have strong clonal formation and multidirectional differentiation potential.Compared with other mesenchymal stem cells,these stem cells have stronger ability of neurogenic differentiation due to their origin in neural crest cell differentiation and migration.VEGFA can not only increase the angiogenesis and regeneration ability,but also has the function of nutrition and nerve protection.At the same time,VEGFA can increase the expression and secretion of nutritional factors in mesenchymal stem cells,improve the microenvironment,and promote the recovery of neurological function.The aim of this study was to explore the effects of VEGFA on the proliferation and neurogenic differentiation of dental pulp stem cells in rats,and to provide a new idea for the treatment of neurological diseases with dental pulp stem cells.MethodsPrimary rat DPSCs were isolated and cultured.The obtained DPSCs were verified by flow cytometry and multidirectional differentiation experiment.Lentiviral vectors carrying VEGFA were constructed to infect DPSCs,and the stable strains were screened and the transfection efficiency was verified.CCK8 assay was used to detect the effect of VEGFA on the proliferation of DPSCs.The expression of nerve markers Nestin,β3-Tubulin and S100β were detected by q PCR,Western Blotting and immunofluorescence assay to evaluate the effect of VEGFA on neurotrophic differentiation of DPSCs in rats.ResultsFlow cytometry showed that the positive rates of CD44 and CD55 were higher than 95%,and the positive rates of CD34 and CD45 were lower than 2%.The results of alizarin red staining and oil red O staining indicated that the cells had the multidirectional differentiation potential of osteogenesis and adipogenesis.It is proved that DPSCs have been successfully extracted and cultured.CCK8 results showed that the proliferation ability of DPSCs in VEGFA-DPSCs group was significantly enhanced at 3,5,7,9 days.After 7 days of induction of neuroblastogenic differentiation medium,q PCR and WB results showed that the expression levels of nerve markers Nestin,β3-Tubulin and S100β were increased after VEGFA overexpression.Immunofluorescence results showed that the fluorescence intensity of the three nerve markers was the strongest in the induced VEGFA-DPSCs group.ConclusionsVEGFA can promote the proliferation of rat dental pulp stem cells.VEGFA can improve the neural differentiation potential of rat dental pulp stem cells and increase the expression of nerve markers.