间充质干细胞治疗单板U型场地运动员肌腱损伤动物模型的建立研究

发布时间：2019-04-20 18:58

【摘要】：伤病问题一直是影响单板U型运动员运动成绩和职业生涯的主要问题,其中肌腱损伤是体育竞技中较为普遍的运动性疾病,导致运动员竞技水平和运动能力不能正常发挥,已成为运动医学领域的急需解决的问题之一。干细胞可以向多胚层分化,在损伤组织修复中也起到了显著的作用。本研究体外培养脐带间充质干细胞(Umbilical cord Mesenchymal Stem Cells,UCMSCs)和牙龈间充质干细胞(Mesenchymal Stem Cells,GMSCs),传代培养到一定体系,检测UCMSCs和GMSCs的生物学特性,比较UCMSCs和GMSCs生物学特性之间的差异,选择更为优质的细胞为肌腱治疗提供研究基础。建立动物损伤模型,深入探究干细胞在肌腱修复过程中所起到的治疗作用,获得以下实验结果:1.使用Ⅱ型胶原酶和胰蛋白酶混合消化羊脐带组织,分离纯化后得到接种在六孔板中,体外培养至31代。用RT-PCR和免疫荧光检测脐带组织来源的干细胞表面标记物表达情况。免疫荧光检测结果,细胞阳性表达的基因为:CD44、CD73、CD90;RT-PCR结果显示:CD29、CD44、CD90表达,不表达的基因为CD34。研究获得细胞的生物学特性,以及诱导细胞向成脂、成骨、成软骨、成肌腱细胞分化。RT-PCR检测其特异性基因均为阳性。2.选择Ⅰ型胶原酶和胰酶混合消化牛牙龈组织,分离出较为纯净的目的细胞,体外培养至20代。分别检测RT-PCR和免疫荧光特异性标记物的表达情况:CD29、CD44、CD105均呈阳性表达。对其生物学特性和多向分化潜能进行研究。3.建立小鼠肌腱损伤模型,用Ⅰ型胶原酶局部注射法损伤小鼠肌腱,采用石蜡病理切片观察肌腱组织的损伤情况,检测小鼠血液中肌酸激酶(CK)含量的变化。将UCMSCs进行荧光标记接种到小鼠损伤部位,冰冻切片检测细胞能向损伤部位集中。进行组织切片和血清学分析肌腱结构和功能得到良好的修复。本研究将围绕以上几点着重介绍间充质干细胞的生物学特性及在肌腱损伤修复过程中所起的重要作用。为干细胞治疗奠定理论基础,为后续临床应用提供实验依据。
[Abstract]:Injuries and injuries have always been the main problems affecting the performance and career of single-board U-type athletes, among which tendon injury is a more common sports disease in sports, which leads to athletes' competitive level and sports ability can not play a normal role. It has become one of the urgent problems in the field of sports medicine. Stem cells can differentiate into polyderm and play a significant role in repair of injured tissues. In this study, umbilical cord mesenchymal stem cells (Umbilical cord Mesenchymal Stem Cells,UCMSCs) and gingival mesenchymal stem cells (Mesenchymal Stem Cells,GMSCs) were cultured in vitro and subcultured to a certain system to detect the biological characteristics of UCMSCs and GMSCs. To compare the difference between the biological characteristics of UCMSCs and GMSCs, we choose better cells to provide the basis for tendon therapy. To establish animal injury model and explore the therapeutic effect of stem cells in tendon repair, the following experimental results are obtained: 1. The umbilical cord tissue of sheep was digested by collagenase 鈪，

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