| Objective: To evaluate the effect of transforming growth factor-beta1 on osteogenic differentiation and the expression of osteoprotegerin/Osteoclast nuclear factor ?B receptor activator ligand in human periodontal ligament stem cells.Method: The recovery of human periodontal ligament stem cells were cultured and passaged and randomly divided into control group(0ng/mL transforming growth factor-beta1)and experimental group(1,10ng/mL,transforming growth factor-beta1).Firstly,CCK-8 detected transforming growth factor-beta1 on cell proliferation;Secondly,the effects of transforming growth factor-beta1 on the osteogenesis of cells was detected by the activity of alkaline phosphatase,alizarin red staining and real-time quantitative PCR;Finally,the effect of transforming growth factor-beta1 on the expression of osteoprotegerin and Osteoclast nuclear factor ?B receptor activator ligand were detected by immunohistochemistry and real-time quantitative PCR in the process of osteogenesis.Results: The experiment of CCK8 showed that transforming growth factor-beta1 can promote cell proliferation and 10ng/mL transforming growth factor-beta1 promotes cell proliferation was the strongest(P<0.05);The experiment of alkaline phosphatase activity assay showed that the activity of alkaline phosphatase in the experimental group was higher than that in the control group,and the activity of 10ng/mL containing transforming growth factor-beta1 in the experimental group was significantly higher than that in the experimental group containing 1ng/mL transforming growth factor-beta1(P<0.05).The experiment of lizarin red staining showed calcification of extracellular matrix.Compared with control group,the calcification of PDLSCs' extracellular matrix in experimental group was significant,and 10ng/mL transforming growth factor-beta1 in the experimental group was significantly more calcified than that in 1ng/mL transforming growth factor-beta1;The experiment of real-time PCR showed experiment group compared with the control group,the expression of cementum attachment protein and osteopontin,osteocalcin,Runt related transcription factor 2,osteoprotegerin and Osteoclast nuclear factor ?B receptor activator ligand were significantly increased(P<0.05)and the ratio of OPG/RANKL were higher than the control group,and the difference was statistically significant(P < 0.05);Immunohistochemistry results showed that the expression of protein of osteoprotegerin and osteoclast nuclear factor ?B receptor activator ligand in periodontal ligament stem cells in experimental group were positive and osteoprotegerin was stronger in the control group than in the control group,while the cytoplasm of the control group was not stained.Osteoprotegerin and osteoclast nuclear factor ?B receptor activator ligand were all negative.Conclusion: TGF-beta 1 can promote bone differentiation of human periodontal ligament stem cells and affect the expression of osteoprotegerin and receptor activator of nuclear factor ?B ligand in this process,up regulation of the ratio of osteoprotegerin/ receptor activator of nuclear factor ?B ligand,which can regulate the regeneration of periodontal bone tissue. |
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