富血小板血浆促猫角膜内皮细胞增殖的研究

Promotive effect of platelet-rich plasma on proliferation of corneal endothelial cells in cats

目的观察富血小板血浆对体外培养猫角膜内皮细胞增殖的影响。方法采用揭膜法与消化法相结合获取猫角膜原代内皮细胞,培养基采用DMEM培养基(含体积分数10%胎牛血清)。采用二次离心法提取富血小板血浆。在完全培养基中加入体积分数分别为5%、10%、20%的富血小板血浆培养角膜内皮细胞,以未加入富血小板血浆的完全培养基为对照。CCK-8试剂盒检测细胞增殖情况,扫描电镜观察细胞形态变化。结果富血小板血浆体外培养角膜内皮细胞时间越长,其促增殖作用越明显,与对照组比较差异均有显著统计学意义意义(均为P<0.01),且随着富血小板血浆含量的增加,角膜内皮细胞增殖越明显,呈剂量依赖性。扫描电镜检测表明,与对照组相比,PRP作用组角膜内皮细胞表面可见丰富的微绒毛,且随着富血小板血浆含量的增加,微绒毛越丰富。结论富血小板血浆能明显促进体外培养猫角膜内皮细胞增殖。

Objective To observe effect of platelet-rich plasma（PRP） on proliferation of corneal endothelial cells（CEC） in cats in vitro.Methods The cat CEC were obtained by separating and treating with enzyme digestion,then cultured in DMEM supplemented with 10% volume fractions fetal bovine serum（FBS）.PRP was extracted by the two-step centrifugation method.The CEC of the second generation were cultured in the complete media containing different concentrations of PRP（5%,10% and 20% volume fractions）,while those cultured in the media without PRP were served as controls.Proliferation of CEC was detected by CCK-8 kit method.The morphological changes of CEC were examined by scanning electron microscope（SEM）.Results Effect of proliferation of the CEC cultured with PRP was obvious with time prolonged,and had statistical difference compared with that of control group（P0.01）.The proliferation of CEC was more obvious with the increase of platelet-rich plasma,and with dose dependent marner.There were more microvilli on the CEC surface in PRP group than control group under scanning electron microscope.The microvini was more with the increase of platelet-rich plasma.Conclusion PRP can obviously promote the proliferation of PRP in cats cultured in vitro.