The Primary Research About Effects Of Salvianolic Acid B On Osteogenic Differentiation Of Human Periodontal Ligament Cells

Objective: Periodontal disease is a common and multiple oral disease which can result in resorption of alveolar bone.Periodontal regeneration research has been the focus of treatment of periodontal disease between Chinese and foreign.To investigate the effects of Salvianolic acid B(Sal B),a major bioactive component of Chinese medicine---Salvia miltiorrhiza,on osteogenic differentiation of human periodontal ligament cells to provide a theoretical basis for the clinical treatment of periodontal disease with salvianolic acid B.Methods:1 The primary human periodontal ligament cells were cultured by tissue block method from young periodontal ligament,then we identify it and the third passage human PDLCs were used to do the experiment.2 Cytotoxicity assay.MTT method was used to observe the proliferation activity of PDLCs with the effects of different concentration of Sal B.3 Experimental grouping.concentration group(Con group),Standard osteogenic induction medium group(OIM group),Standard osteogenic induction medium group+ salvianolic acid B group(OIM + Sal B group): the concentration of Sal B was 0.1?M,0.5?M,1?M,5?M.4 Measurement of Alkaline phosphatase(ALP)activity.ALP activity was measured of PDLCs after cultured for 7 days.5 ALP staining of PDLCs.ALP staining was measured of PDLCs after cultured for 7 days.6 Mineralized nodule staining.alizarin red was used to stain the cells after cultured for 20 days.7 Real-time PCR.The expression of OCN mRNA in human periodontal ligament cells was detected by Real-time PCR after cultured for 10 days.Results:1 PDLCs were successfully cultured and identified.2 The results of cytotoxicity test showed that the salvianolic acid B had no toxicity for human periodontal ligament cells in concentration of 0.1?M,0.5?M,1?M,5?M.3 ALP activity of PDLCs was improved in treatment group with different concentrations of Sal B(0.5?M?1?M?5?M).Compared with OIM group,statistically significant differences were found(P<0.05).4 The results of ALP activity staining showed that the degree of cytoplasm blue staining of OIM + Sal B(0.1?M,0.5?M,1?M,5?M)was higher than that of OIM group.5 The results of Mineralized nodules staining showed that When the concentration of Sal B was 0.5M,1?M,5?M the mineralized nodules in cells were significantly higher than the OIM group.6 Real-time PCR results: the OCN mRNA expression can be increased with the concentration of salvianolic acid B in 0.5?M,1?M,5?M,Compared with OIM group,statistically significant differences were found(P<0.05).Conclusion:Appropriate concentration of Sal B can improve the osteogenic differentiation of PDLCs.Sal B has a broad application prospects and value in the periodontal tissue regeneration and repair.