【摘要】：傷病問題一直是影響單板U型運動員運動成績和職業(yè)生涯的主要問題,其中肌腱損傷是體育競技中較為普遍的運動性疾病,導(dǎo)致運動員競技水平和運動能力不能正常發(fā)揮,已成為運動醫(yī)學(xué)領(lǐng)域的急需解決的問題之一。干細(xì)胞可以向多胚層分化,在損傷組織修復(fù)中也起到了顯著的作用。本研究體外培養(yǎng)臍帶間充質(zhì)干細(xì)胞(Umbilical cord Mesenchymal Stem Cells,UCMSCs)和牙齦間充質(zhì)干細(xì)胞(Mesenchymal Stem Cells,GMSCs),傳代培養(yǎng)到一定體系,檢測UCMSCs和GMSCs的生物學(xué)特性,比較UCMSCs和GMSCs生物學(xué)特性之間的差異,選擇更為優(yōu)質(zhì)的細(xì)胞為肌腱治療提供研究基礎(chǔ)。建立動物損傷模型,深入探究干細(xì)胞在肌腱修復(fù)過程中所起到的治療作用,獲得以下實驗結(jié)果:1.使用Ⅱ型膠原酶和胰蛋白酶混合消化羊臍帶組織,分離純化后得到接種在六孔板中,體外培養(yǎng)至31代。用RT-PCR和免疫熒光檢測臍帶組織來源的干細(xì)胞表面標(biāo)記物表達(dá)情況。免疫熒光檢測結(jié)果,細(xì)胞陽性表達(dá)的基因為:CD44、CD73、CD90;RT-PCR結(jié)果顯示:CD29、CD44、CD90表達(dá),不表達(dá)的基因為CD34。研究獲得細(xì)胞的生物學(xué)特性,以及誘導(dǎo)細(xì)胞向成脂、成骨、成軟骨、成肌腱細(xì)胞分化。RT-PCR檢測其特異性基因均為陽性。2.選擇Ⅰ型膠原酶和胰酶混合消化牛牙齦組織,分離出較為純凈的目的細(xì)胞,體外培養(yǎng)至20代。分別檢測RT-PCR和免疫熒光特異性標(biāo)記物的表達(dá)情況:CD29、CD44、CD105均呈陽性表達(dá)。對其生物學(xué)特性和多向分化潛能進(jìn)行研究。3.建立小鼠肌腱損傷模型,用Ⅰ型膠原酶局部注射法損傷小鼠肌腱,采用石蠟病理切片觀察肌腱組織的損傷情況,檢測小鼠血液中肌酸激酶(CK)含量的變化。將UCMSCs進(jìn)行熒光標(biāo)記接種到小鼠損傷部位,冰凍切片檢測細(xì)胞能向損傷部位集中。進(jìn)行組織切片和血清學(xué)分析肌腱結(jié)構(gòu)和功能得到良好的修復(fù)。本研究將圍繞以上幾點著重介紹間充質(zhì)干細(xì)胞的生物學(xué)特性及在肌腱損傷修復(fù)過程中所起的重要作用。為干細(xì)胞治療奠定理論基礎(chǔ),為后續(xù)臨床應(yīng)用提供實驗依據(jù)。
[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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