The Effect Of Saliva Miltiorrhiza Bugge, RhBMP-2 And RhTGF-β1 On Human Periodontal Ligament Fibroblasts In Vitro

The key of periodontal tissue regeneration is the proliferation and differentiation of human periodontal ligament fibroblasts(HPDLFs).The cells in pathological region are lack,so stimulating the proliferation and differentiation of HPDLFs is the key problem of research on periodontal tissue regeneration.The investigation in last years showed that Saliva Miltiorrhiza Bugge can improve the minimum-cycle,intervene the inflammation,adjust the synthesis of growth factors,and promote the tissue regeneration.The animal experiment suggested that Saliva Miltiorrhiza Bugge combined with biomembrane in GTR can promote better periodontal tissue regeneration.The development and proper application of growth factors play an very important role in promoting the development of the seed cell's potential in periodontal tissue engineering.The objective of this study is to invstigate the effect of Saliva Miltiorrhiza Bugge on HPDLFs in vitro.HPDLFs were cultured with different concentration of Saliva Miltiorrhiza Bugge,the proliferation and differentiation of HPDLFs were observed.Objective:To study the effects of Saliva Miltiorrhiza Bugge(SMB),bone morphogenetic protein-2(rhBMP-2),and transforming growth factor-β1(rhTGF-β1)on the proliferation and differentiation of human periodontal ligament cells(HPDLFs) respectively and select their optimal density.Methods:The first premolars extracted due to orthognathous treatment were selected and the one-third PDL tissures of fang's intermediate were scraped as samples.The fifth generation of the cells was used for the experiment.After subcultivation,the DMEM medium was supplemented with 5 concentrations of SMB45mg/L, 90mg/L,180 mg/L,360 mg/L,720mg/L,rhBMP-2 -25μg/L,50μg/L, 100μg/L,200μg/L,400μg/L.and rhTGF-β1-2.5μg/L,5μg/L,10μg/L,20μg/L, 40μg/L.The cells were cultured for 1,3,5,7 days respectively.The cells number was measured,and select the optimal density of SMB,rhBMP-2 and rhTGF-β.The cells number,alkaline phosphoric activity(ALP activity)and the amount of OCN of the cell's was mesured after 5-,7-,12- and 14- day culture with their optimal density respectively.PDLFs culture technique,MTT assay and enzyme kinetics methods were used to observe dynamically the effects of gradient contents of SMB,rhTGF-β1 and rhBMP -2 on the proliferation and differentiation of PDLFs respectively.Result:SMB,rhBMP-2 and rhTGF-β1 can promote the proliferation and differentiation of PDLFs。The optimal density of them is SMB(360rag/L),rhBMP-2 (200μg/L)and rhTGF-β1(5μg/L)).All of them dynamically acted on HPDLFs from 1 to 7 days and indicated continuously promoting proliferation and differentiation trend. SMB(360mg/L),rhBMP-2(200μg/L)and rhTGF-β1(5μg/L)can strengthened the ALP activity and OCN activity。There are no differences(P＜0.05)among SMB(360mg/L), rhBMP-2(200μg/L),and rhTGF-β1(5μg/L)Conclusion:SMB,rhBMP-2 and rhTGF-β1 can promote the proliferation and differentiation of.PDLFs.At the optimal density,There are no differences among them.(P＜0.05)...