The Effect Of MiR-30a Regulating Runx2 On The Osteogenic Differentiation Of Human Periodontal Ligament Stem Cells Under Inflammatory Microenvironment

ObjectiveTo investigate the effect of micro RNA-30a(miR-30a)regulating Runt-related transcription factor 2(Runx2)in inflammatory microenvironment on the osteogenic differentiation of human periodontal ligament stem cells.MethodHuman periodontal ligament stem cells(hPDLSCs)were cultured in vitro and randomly divided into control group,model group,miR-30 a mimics group,miR-30 a mimics negative control group,miR-30 a inhibitor group and miR-30 a inhibitor negative control group,in addition to the control group,the other groups used lipopolysaccharide to induce the inflammatory microenvironment of the cells,constructed the inflammatory model in vitro,and then treated them in groups and induced their osteogenic differentiation,the alkaline phosphatase(ALP)and alizarin red staining were used to detect the osteogenic differentiation;enzyme linked immunosorbent assay(ELISA)was used to detect the levels of TNF-α and IL-6 in the supernatant of each group;the expressions of miR-30 a,Runx2 mRNA were detected by real-time fluorescence quantitative analysis(qRT-PCR);and the expressions of Runx2,osteopontin(OPN)and osteocalcin(OCN)proteins were detected by Western blot.ResultCompared with the control group,the expression of miR-30 a and the levels of TNF-α and IL-6 in the supernatant of the model group were higher(P <0.05),the expression level of Runx2 mRNA,relative proportion of ALP positive cells,relative proportion of mineralized nodule,expression levels of Runx2,OPN and OCN proteins were lower(P<0.05).Compared with the model group,the expression of miR-30 a and the levels of TNF-α and IL-6 in the supernatant of miR-30 a mimics group were higher(P<0.05),the expression level of Runx2 mRNA,relative proportion of ALP positive cells,relative proportion of mineralized nodule,expression levels of Runx2,OPN and OCN proteins were lower(P<0.05);the expression of miR-30 a and the levels of TNF-α and IL-6 in the supernatant of miR-30 a inhibitor group were lower(P<0.05),the expression level of Runx2 mRNA,relative proportion of ALP positive cells,relative proportion of mineralized nodule,expression levels of Runx2,OPN and OCN proteins were higher(P<0.05);there was no significant change in cell indexes of miR-30 a mimics negative control group and miR-30 a inhibitor negative control group(P>0.05).ConclusionThe miR-30 a is highly expressed in hPDLSCs in inflammatory microenvironment,which can down-regulate the expression of Runx2,promote the progress of inflammation,and then inhibit the osteogenic differentiation of hPDLSCs.