【摘要】 目的 分析Choukroun′s富血小板纤维蛋白(PRF)对人成骨细胞增殖分化及ERK1/2-Runx2通路的影响。方法 组织块法分离人成骨细胞，设置对照组、PRF1组和PRF2组，分别用不含PRF、含1×PRF浸出液和2×PRF浸出液的DMEM完全培养基培养。检测细胞增殖、碱性磷酸酶(ALP)活性、I型胶原表达及细胞矿化能力，Western blot法测定细胞外调节蛋白激酶1/2-Runt相关转录因子2(ERK1/2-Runx2)信号通路蛋白表达情况。结果 与对照组比较，PRF1组和PRF2组MTT实验A值及ALP活性均升高，且PRF2组高于PRF1组(P<0.05);与培养1天比较，培养3、7天时三组MTT实验A值均升高，且培养7天高于培养3天(P<0.05)。培养7天，PRF1组和PRF2组I型胶原平均灰度值高于对照组，PRF2组高于PRF1组(P<0.05)。培养14、21天，PRF1组和PRF2组钙结节积分光密度高于对照组，PRF2组高于PRF1组(P<0.05);培养21天三组钙结节积分光密度均高于14天(P<0.05)。与对照组比较，PRF1组和PRF2组p-ERK1/2、Runx2蛋白表达升高，且PRF2组高于PRF1组(P<0.05)。结论 PRF可能通过激活ERK1/2-Runx2信号通路参与人成骨细胞增殖分化过程。更多还原

【Abstract】 Objective To analyze the effects of Choukroun′s platelet-rich fibrin(PRF) on the proliferation, differentiation and ERK1/2-Runx2 pathway of human osteoblasts.Methods Human osteoblasts were isolated by tissue block method, and divided into a control group, a PRF1 group and a PRF2 group. The cells were cultured with DMEM complete medium without PRF, with 1×PRF leaching solution, and 2×PRF leaching solution, respectively. Cell proliferation, alkaline phosphatase(ALP) activity, collagen type I expression and cell mineralization capacity were detected. The protein expression of extracellular regulated protein kinase 1/2-Runt-related transcription factor 2(ERK1/2-Runx2) signaling pathway was determined by Western blot.Results The A value and ALP activity of the MTT experiment in the PRF1 group and the PRF2 group were increased higher than those in the control group, and the PRF2 group was higher than the PRF1 group(P<0.05). Compared with the 1 st day of culture, the A values of the three groups in the MTT experiment were increased on the 3 rd and the 7 th day of culture, and the 7 th day of culture was higher than that of the 3 rd day of culture(P<0.05). After 7 days, the average gray value of collagen type I in the PRF1 and PRF2 groups were higher than those in the control group, and the PRF2 group was higher than the PRF1 group(P<0.05). After 14 and 21 days of culture, the integral optical density of calcium nodules in the PRF1 and PRF2 groups were higher than those in the control group, and the PRF2 group was higher than PRF1 group(P<0.05). The integral optical density of calcium nodules in the three groups at the 21 st day was higher than that at the 14 th day(P<0.05). The expressions of p-ERK1/2 and Runx2 proteins in the PRF1 and PRF2 groups were higher than those in the control group, and the PRF2 group was higher than the PRF1 group(P<0.05).Conclusion PRF may participate in the proliferation and differentiation of human osteoblasts by activating the ERK1/2-Runx2 signaling pathway.更多还原