富血小板血浆对巨噬细胞表型的作用及其影响因素研究(英文)

Efects of Platelet-Rich Plasma on Macrophage Phenotype and Its Influencing F actors

目的:评估临床常用的几种血小板激活剂,即凝血酶、ADP和CaCl_(2)活化的富血小板血浆(PRP)上清对巨噬细胞表型的影响并探究储存时间对PRP功能的影响。方法:采用含不同激活剂或不同储存时间的活化PRP上清的培养基体外培养人单核衍生巨噬细胞,流式细胞仪检测巨噬细胞表面标记分子的表达,ELISA方法检测PRP上清中的生长因子浓度。结果:培养24 h后,凝血酶和CaCl_(2)活化PRP上清刺激的巨噬细胞CD86的表达水平均明显高于PRP组(P<0.05),且CaCl_(2)活化PRP上清增加了巨噬细胞CD163的表达。而对于不同激活剂组比较,CaCl_(2)活化组的巨噬细胞CD163的表达明显高于凝血酶和ADP组(P<0.05)。ELISA结果显示,在-80℃储存大于20个月和10-20个月的PRP在CaCl_(2)活化后,其上清中FGF(P<0.001)和EGF(P<0.05)的浓度明显高于储存小于10个月组,且两组上清处理的巨噬细胞CD86(P<0.01)、CD163(P<0.001)和CD206(P<0.001)表达均明显升高。结论:不同激活剂活化的PRP对巨噬细胞的表型影响不同。同时,储存时间也会影响PRP的生长因子浓度和作用效果。

Objective:To evaluate the effects of platelet-rich plasma(PRP)supernatants with different activ ation methods and storage time on human monocyte-derived macrophages phenotype and explore the possible mechanism.Methods:Human monocyte-derived macrophages were cultured in vitro with PRP or activated PRP supernatants activated with different activators.The expression of marker molecules on the suface of macrophages was detected by flow cytometry,and the concentration of growth factors in PRP supernatants was detected by ELISA.Results:After 24 h of coculture,the expression level of CD86 in macrophages stimulated by PRP supernatant(thrombin and CaCl_(2) activated)was significantly higher than that by PRP group(P<0.05),and the expression of CD163 in macrophages was increased by CaCl_(2) activated PRP supematant.Compared with different activator groups,the expression of CD163 in macrophages of CaCl_(2) activated group was significantly higher than that of thrombin and ADP groups(P<0.05).ELISA results showed that the concentrations of FGF(P<0.001)and EGF(P<0.05)in the supematant of PRP stored at-80 C for more than 20 months and 10-20 months were significantly higher than those in the group stored at less than 10 months after CaCl_(2) activation,and the expressions of CD86(P<0.01),CD163(P<0.001)and CD206(P<0.001)in macrophages cocultured with the supematant of the two groups were significantly increased.Conclusion:PRP activated by different activators has different effects on the phenotype of macrophages.Meanwhile,the storage time will also affect the growth factor concentration and effect of PRP.