【摘要】 目的:探索不同浓度富血小板血浆(platelet-rich plasma,PRP)对人牙周膜细胞体外增殖和碱性磷酸酶(alkaline phosphatase,ALP)活性的影响。方法:两步离心法制备PRP,用酶联免疫吸附法检测血浆、PRP、激活后PRP上清液中转化生长因子-β1(transforming growth factors-β1,TGF-β1)和血小板衍生生长因子(platelet-derived growth factor-AB,PDGF-AB)的水平。分别用2%、5%、10%、20%激活后的PRP作用于人牙周膜细胞,以DMEM培养液为阴性对照。在作用24h和72h后,用细胞计数试剂盒检测细胞增殖状况,以对硝基磷酸二钠为底物,检测细胞ALP的活性。用SPSS10.0软件包中的方差分析和配对t检验进行统计学分析。结果:激活后的PRP上清中,TGF-β1和PDGF-AB的水平均高于未经激活的PRP和血浆。各浓度PRP促细胞增殖和ALP活性的作用均显著强于阴性对照组(P<0.001);各浓度PRP72h时促细胞增殖和ALP活性的作用均高于24h时(P<0.01);不同浓度PRP组间存在显著差异(P<0.001),PRP浓度从2%渐增至10%时,细胞增殖和ALP活性随之增加;当PRP浓度达到20%时,促细胞增殖作用开始下降,而促ALP活性作用继续增强。结论:在1～3d内,PRP对人牙周膜细胞的体外增殖和ALP活性均有促进作用,并在一定范围内呈剂量依赖关系。更多还原

【Abstract】 PURPOSE: To assess the effect of different concentrations of platelet-rich plasma (PRP) on cell proliferation and alkaline phosphatase (ALP) activity of human periodontal ligament cells (hPDLCs) in vitro. METHODS: PRP was obtained from volunteer donors using two-step centrifugation. TGF-β1 and PDGF-AB levels in activated PRP,PRP and plasma were evaluated by enzyme-linked immunosorbent assay. hPDLCs were exposed to various concentrations of PRP (2%,5%,10% and 20%) and DMEM (negative control),respectively. After 24 and 72 hours,cell proliferation was evaluated using Cell Counting Kit-8 assay. Alkaline phosphatase (ALP) activity of hPDLCs was evaluated by a p-nitrophenyl phosphate disodium assay. Statistical analysis was performed using one-way ANOVA and Student’s t test in SPSS10.0.RESULTS: TGF-β1 and PDGF-AB levels were highest in activated PRP. The effect of various concentrations of PRP on cell proliferation and ALP activity was significantly greater than that of negative control (P<0.001). The effect of PRP was significantly greater at 72-hour compared with that at 24-hour (P<0.01),with significantly difference among various PRP concentrations (P<0.001). Cell proliferation and ALP activity increased when the concentration of PRP increased from 2% to 10%. The maximum effect on cell proliferation was achieved with 10% PRP; 20% PRP resulted in a reduced cell proliferation. However,ALP activity was greatest with 20% PRP. CONCLUSION: PRP preparations exert a dose-dependent effect on hPDLCs proliferation and ALP activity in a certain extent in vitro at 24-hour and 72-hour. Supported by the Capital Medical Development Fund (Grant No. 2005-2007).更多还原