The Effect Of Prp On Periodontal Ligament Flbroblasts In Different Conditions

Objectives: The aim of the study was to investigate the in vitro effects of platelet-richplasma on human periodontal ligament fibroblasts(PDLFs) proliferation ,migrationand human periodontal ligament fibroblasts proliferation co-culture with human bonemarrow stroma cells(BMSCs).Provide the data for the application of PRP inperiodontal tissue regeneration treatment .Methods:1. HPDLFs were isolated and cultured using tissue culture In vitro.Determine the biological sources by detecting the expression of vimentin. HBMSCswere separated and cultured using whole bone marrow culture。PRP was obtainedfrom Jiangsu province blood center.2. HPDLFs were exposed to various concentrations of PRP(1%,5%,10%,20%,30%,50%). HPDLFs proliferation was evaluated using an MTTproliferation assay. Migration was determined by assessing HPDLFs response tovarious concentrations of PRP(1%,5%,10%,20%,30%,50%) with Transwell chamber.10% newborn calf serum was positive control and 0% newborn calf serum wasnegative control.3. HPDLFs and HBMSCs were seeded in transwell system at 1:1ratio. 10% newborncalf serum was positive control and 0% newborn calf serum was negative control.HPDLFs cultured alone were negative control for co-culture. The co-cultures wereincubated for 7 days. The proliferation rates of HPDLFs were assessed by MTTassays. ResultsResults:1.HPDLFs separated from tissue culture expressed vimentin, Identify that the cellswere derived from mesoderm.HBMSCs separated from whole bone marrow werefibroblast-like appearance, which had good proliferation in vitro.2. At 3, 5and 7days, the various concentrations of PRP groups compared with thenegative control group significantly stimulated cell proliferation of HPDLFs (P<0.01). The increase in cell proliferation treated by 20% PRP was statistically higherthan with other PRP concentrations (p < 0. 01). The data suggested that PRPtreatment induced a strong time-dependent increase of HPDLFs proliferation in the 7days culture.The various PRP concentrations significantly enhanced migration compared with thenegative control of 0% newborn calf serum.Especially,concentrations of 10% PRPshowed the most significant stimulation among them(p < 0.01).3. Following 7 days of exposure to the various concentrations of PRP, there was astatistically significant increase in proliferation of HPDLFs.A significant differencebetween platelet-rich plasma-treated cells and negative control groups wereobserved at 7 day(p <0.01), and co-culture group compared with non-co-culturegroup had better proliferative capacity (p <0.01).The increase of proliferation inducedby 10% PRP was statistically higher than other concentrations of PRP(p <0.01).Conclusions: Platelet-rich plasma can exert a positive effect on human periodontalligament fibroblasts function. But this effect with higher concentrations resulted in areduction outcomes.