Effect Of Overexpression Of Transcription Factor And High Magnesium Ion Concentration Environment On Osteogenic Differentiation Of Tissue Engineering Seeding Cells

PURPOSE To explore the effect of overexpression of Runx2 and Osterix(OSX)genes on the osteogenic differentiation of human umbilical vein endothelial cells(HUVECs)and to explore the effect of the high magnesium ion environment on the osteogenic differentiation of rat Bone Mesenchymal Stem Cells(BMSCs).METHODS Overexpression Runx2 and osterix lentiviral vectors were transfected into HUVECs respectively,and the BMSCs were cultured in high magnesium ion environment.The osteogenic potential and the mechanism of transfected cells or BMSCs cultured in high magnesium ion concentration environment were identified by alkaline phosphatase(alkaline phosphatase,ALP)staining and ALP activity.Furthermore,q PCR,Western blot and immune-fluorescence staining were performed to detect the expression of osteogenic genes and proteins in HUVECs and BMSCs.RESULTS 1.The overexpression of Runx2 gene was beneficial for the osteogenic differentiation of HUVECs,and the gene expression level of Runx2,OSX,ALP,bone sialoprotein(BSP),osteopontin(OPN),osteocalcin(OCN),and the up-regulated protein level of OPN and OCN was also observed.While the overexpression of OSX can not promote the osteogenic differentiation of HUVECs.2.The ALP activity of BMSCs increased in the presence of high magnesium ion concentration,and the transcription level of Runx2,ALP,OCN were up-regulated,and the expression of OCN protein was increased.In addition,the phosphorylation level of ERK protein in the cells increased in the presence of high magnesium ion concerntration.CONCLUTIONS The overexpression of Runx2 could promote the osteogenic differentiation of HUVECs.And the high magnesium ion concentration environment could promote the osteogenic differentiation of rat BMSCs and it was actived through ERK pathway.