富血小板血浆对人脂肪间充质干细胞增殖及成骨分化能力的影响  

Effect of Platelet-rich Plasma on the Proliferation and Osteogenic Differentiation of Human Adipose-derived Mesenchymal Stem Cells in Vitro

作　　者：王悦[1] 杨旭芳[2,3] 周延民[1] 赵静辉[1] 朱喆[2] 马英智[2,4] 

机构地区：[1]吉林大学口腔医院种植中心,吉林长春130021 [2]吉林大学基础医学院病理生物学教育部重点实验室 [3]牡丹江医学院病理生理学教研室 [4]空军航空大学训练基地门诊部

出　　处：《口腔医学研究》2011年第9期786-789,共4页Journal of Oral Science Research

基　　金：吉林省科学技术厅项目(编号:200804423)

摘　　要：目的:探讨富血小板血浆(PRP)对人脂肪间充质干细胞(hADSCs)增殖、分化及成骨能力的影响。方法:脂肪组织体外分离获得hADSCs,2次离心制备PRP,氯化钙加人凝血酶激活PRP。倒置相差显微镜观察成骨诱导实验中PRP对细胞增殖、分化状况的影响,钙-钴法检测PRP作用下hADSCs矿化结节形成,碱性磷酸酶(ALP)检测PRP对细胞活性及分化能力的影响。CCK-8法检测成骨诱导组和非成骨诱导组加PRP培养后2、4、6、8、16d细胞活性变化。结果:成骨诱导实验中50%PRP30μL组细胞密集、数量最多,且有剂量依赖性;钙-钴法检测见PRP原液组细胞染色最深,且有浓度依赖性;碱性磷酸酶显色见PRP原液组细胞活性最强,染色最深。CCK-8检测显示无论有无成骨诱导,各浓度PRP均可促进细胞增殖,且有浓度依赖性。结论:适宜浓度的PRP可明显促进hADSCs增殖,并有浓度及剂量依赖性。Objective： To evaluate the effect of platelet-rich plasma（PRP） on proliferation and osteogenic differentiation of human adipose-derived mesenchymal stem cells（hADSCs） in vitro.Methods： hADSCs were separated from an adult donor.PRP was prepared by twice centrifugation method.CaCl2 combined with thrombin was used to activate PRP.In the osteogenesis induction experiment the morphology of cells was studied with a phase contrast microscope.The alkaline phosphatase（ALP） activity of hADSCs in different concentrations PRP groups was determined by calcium-cobalt method and BCIP/NBT alkaline phosphatase color development kit was used,respectively.The proliferation of hADSCs with and without osteogenic induction culture medium in various concentrations PRP groups was detected by CCK-8 assay in 2nd,4th,6th,8th and 16th day.Results： The microscope investigation showed that 50% PRP 30μL group improved cell proliferation higher than other groups.The significant increase in ALP was observed when added original liquid PRP.The CCK-8 showed that mean A450 of original liquid PRP was higher than other groups whichever with or without osteogenic induction.Conclusion： These data suggest that PRP has a positive dose-depend and concentration-depend influence on hADSCs proliferation.

关 键 词：人脂肪间充质干细胞 富血小板血浆 成骨诱导 增殖 分化 

分 类 号：R782[医药卫生—口腔医学]