| Objective:To evaluate the effects of tumor necrosis factor-a on osteogenic differentiation and Notch signaling pathway of periodontal ligament stem cell,to investigate the regulatory role of Notch signaling pathway on the osteogenic differentiation of periodontal ligament stem cell under the influence of tumor necrosis factor-a.Methods:periodontal ligament stem cell was obtained by enzyme digestion and tissue block method,the expression of stem cell surface markers CD 105,CD90,CD 146,CD45 and CD31 were detected by Fluorescence activated cell sorter,periodontal ligament stem cell was divided into experimental group(tumor necrosis factor-a lOng/ml)and control group(tumor necrosis factor-a Ong/ml),The proliferation ability of periodontal ligament stem cell was detected by CCK-8;The effect of tumor necrosis factor-a on the osteogenic ability of periodontal ligament stem cell was testde by alkaline phosphatase activity alizarin red staining and quantitative real-time PCR;The Notch signal pathway receptor Notch1,Notch2;ligand JAG1,JGA2;downstream gene Hes1,DLL1 expression changes were detected by quantitative real-time PCR.Results:Fluorescence activated cell sorter profiling showed that periodontal ligament stem cell were strongly positive for CD105,CD90 and CD146,but negative for CD45 and CD31;The results of CCK-8 showed that tumor necrosis factor-a could promote the proliferation of periodontal ligament stem cell(P?0.05);The results showed that the activity of ALP in the experimental group was lower than that in the control group(P<0.05);Alizarin red staining showed that compared with the control group,the mineralized nodules of the experimental group were smaller and the amount of the formation was also reduced;Quantitative real-time PCR results showed that compared with the control group,the mRNA expression of osteogenic marker genes cementum attachment protein,osteopontin,runt-related transcription factor 2 significantly decreased(P?0.05).The expression levels of Notch1,Notch2,JAG 1,JGA2 and Hes-1 were significantly lower than control group(P?0.05),while the expression levels of Notch3 and DLL1 were increased in Notch signaling pathway related molecules(P<0.05).Conclusion:tumor necrosis factor-a can promote the proliferation of periodontal ligament stem cell and inhibit the differentiation of bone and inhibit the expression of Notch signaling pathway,which shows that the Notch signaling pathway has a certain regulatory effect on osteogenic differentiation of periodontal ligament stem cell. |
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