富血小板血浆(PRP)凝胶联合TGF-β3对肌腱干细胞成软骨分化影响的实验研究  

Study on effect of PRP gel combined with TGF-β3 on chondrogenic differentiation of tendon stem cells

作　　者：罗毅 汪敏加[2] 苏全生 何本翔[2] 邓友章 高伟强[4] 姜河[4] LUO Yi;WANG Min-jia;SU Quan-sheng;HE Ben-xiang;DENG You-zhang;GAO Wei-qiang;JIANG He(Affiliated Hospital of Chengdu Institute of Physical Education,Chengdu 610041,China;College of Sports and Health,Chengdu Institute of Physical Education,Chengdu 610041,China;Chengdu University of Traditional Chinese Medicine,Chengdu 610075,China;Chengdu First People's Hospital,Chengdu 610041,China)

机构地区：[1]成都体育学院附属医院博士后工作站,四川成都610041 [2]成都体育学院运动与健康学院,四川成都610041 [3]成都中医药大学,四川成都610075 [4]成都市第一人民医院骨科,四川成都610041

出　　处：《广西大学学报（自然科学版）》2019年第6期1781-1789,共9页Journal of Guangxi University（Natural Science Edition）

基　　金：国家自然科学基金资助项目(BK2010572);四川省中医药管理局课题项目(2018JC034);成都体育学院运动医学与健康研究所/2017年创新课题项目(CX17AO4)

摘　　要：探究PRP凝胶联合转化生长因子-β3(TGF-β3)对肌腱干细胞成软骨分化的影响。方法:体外分离培养人跟腱来源肌腱干细胞hTSCs,经表面标志物检测鉴定后,制备富血小板血浆(PRP)凝胶-hTSCs细胞复合物,以CCK-8法检测PRP凝胶的细胞相容性;体外培养的hTSCs随机分为对照组(control)、TGF-β3组、PRP凝胶组、PRP凝胶+TGF-β3组四组,经TGF-β3、PRP凝胶处理并诱导其成软骨分化,甲苯胺蓝染色检测成软骨分化情况,以实时荧光定量(qRT-PCR)及蛋白免疫印迹法(Western blot)检测成软骨分化标志Sox9、Ⅱ型胶原(CollagenⅡ)的表达。PRP凝胶具有良好的细胞相容性,并对hTSCs有一定的促增殖作用。与control组相比,TGF-β3组、PRP凝胶组、PRP凝胶+TGF-β3组细胞成软骨细胞比例、Sox9、CollagenⅡ表达均增加(P<0.05);与TGF-β3组、PRP凝胶组分别相比,PRP凝胶+TGF-β3组细胞成软骨细胞比例、Sox9、CollagenⅡ表达均增加(P<0.05)。PRP凝胶联合TGF-β3促进肌腱干细胞成软骨分化优于单独使用。To explore the effect of PRP gel combined with transforming growth factor-β3(TGF-β3)on chondrogenic differentiation of tendon stem cells.Human Achilles tendon stem cells(hTSCs)were isolated and cultured in vitro,after the identification of surface markers,platelet rich plasma(PRP)gel-hTSCs cell complex was prepared,and the cell compatibility of PRP gel was detected by CCK-8.HTSCs cultured in vitro were randomly divided into four groups:control group,TGF-β3 group,PRP gel group and PRP Gel+TGF-β3 group.The TGF-β3 and PRP gel were used to induce chondrogenesis.The cartilage differentiation was detected by toluidine blue staining.The real-time fluorescence quantitative(qRT-PCR)and Western blot were used to detect the expressions of Sox9 and CollagenⅡ,markers of chondrogenic differentiation.PRP gel had good cytocompatibility and had certain promoting effect on hTSCs.Compared with group control,the proportion of chondrocytes,Sox9 and CollagenⅡincreased in TGF-β3 group,PRP gel group,PRP Gel+TGF-β3 group(P<0.05).Compared with TGF-β3 group and PRP gel group,the proportion of chondrocytes,Sox9 and CollagenⅡincreased in PRP Gel+TGF-β3 group(P<0.05).The effect of PRP gel combined with TGF-β3 on promoting chondrogenesis of tendon stem cells is better than that of single use.

关 键 词：PRP凝胶 TGF-Β3 联合 肌腱干细胞 成软骨分化 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]