转化生长因子β及周期性拉伸应变条件下骨髓间充质干细胞向软骨样细胞的分化  

Differentiation of bone marrow mesenchymal stem cells into chondrocyte-like cells under the conditions of transforming growth factor beta and cyclical tensile strain

作　　者：郝耀[1] 乔梁[1] 郝永壮[1] 向川[1] 

机构地区：[1]山西医科大学第二医院骨科,山西省太原市030001

出　　处：《中国组织工程研究》2014年第28期4429-4436,共8页Chinese Journal of Tissue Engineering Research

摘　　要：背景:研究表明转化生长因子β对骨髓间充质干细胞的软骨方向分化具有显著的诱导作用。周期性拉伸应变可以模拟软骨细胞在体内的力学环境,对细胞的增殖和分化起着重要的调节作用。目的:探讨转化生长因子β及周期性拉伸应变在诱导骨髓间充质干细胞向软骨样细胞分化过程中是否具有协同作用。方法:取2月龄新西兰大白兔10只,骨穿针穿入股骨髓腔内,抽取骨髓3.0-4.0 mL并分离培养骨髓间充质干细胞,传至3代后随机分为4组,分别为空白组、转化生长因子β组、周期性拉伸应变组以及周期性拉伸应变+转化生长因子β组,作用1,3,6 d后取出相应细胞,番红O染色观察大体形态,阿尔新蓝染色检测糖胺聚糖水平,ELISA检测上清液基质金属蛋白酶13及基质金属蛋白酶组织抑制剂1水平,RT-PCR检测Ⅱ型胶原、基质金属蛋白酶13及基质金属蛋白酶组织抑制剂1 mRNA相对表达量。结果与结论:番红O染色可见细胞呈长梭形或不规则三角形样改变,各实验组较空白组细胞数量及基质分泌增多。作用第3天时转化生长因子β组、周期性拉伸应变+转化生长因子β组上清液糖胺聚糖水平较空白组均升高(P<0.05),周期性拉伸应变+转化生长因子β组Ⅱ型胶原mRNA相对表达量较空白组增高(P<0.05)。结果提示转化生长因子β及周期性拉伸应变均可诱导骨髓间充质干细胞向软骨细胞分化,二者具有明显的协同作用。BACKGROUND：Transforming growth factor-βhas been shown to exert an obvious induction effect on the differentiation of bone marrow mesenchymal stem cells into chondrocytes. Cyclical tensile strain simulates mechanical environment of chondrocytes in the body, and plays an important regulatory role in cellproliferation and differentiation. OBJECTIVE：To discuss the synergy of transforming growth factor-βand cyclical tensile strain in inducing the differentiation of bone marrow mesenchymal stem cells into chondrocyte-like cells. METHODS：A total of 10 2-month-old New Zealand rabbits were selected. Bone needle was used to penetrate the medul ary cavity of bone. 3.0-4.0 mL of bone marrow was extracted for isolation and culture of bone marrow mesenchymal stem cells. Passage 3 cells were randomly assigned to four groups：blank, transforming growth factor-β, cyclical tensile strain and cyclical tensile strain＋transforming growth factor-βgroups. After 1, 3 and 6 days, cells were obtained. General morphology was observed using safranin O staining. Glycosaminoglycan levels were detected by alcian blue staining. Matrix metal oproteinase-13 and tissue inhibitor of metal oproteinase-1 levels in supernatant were measured using ELISA. Type II col agen, matrix metal oproteinase-13 and tissue inhibitor of metal oproteinase-1 mRNA relative expression was detected using RT-PCR. RESULTS AND CONCLUSION：Safranin O staining showed fusiform or irregular triangular cells. cellnumber 〈br〉 and matrix secretion increased in each experimental group than in blank group. Glycosaminoglycan levels in the supernatant were greater in the transforming growth factor-βand cyclical tensile strain＋transforming growth factor-βgroups than in the blank group （P〈0.05）. Type II col agen mRNA relative expression was higher in the cyclical tensile strain＋transforming growth factor-βgroup than in the blank group （P〈0.05）. Results indicated that transforming growth factor-βand cyclical tensile strain could induce the differenti

关 键 词：干细胞 骨髓干细胞 骨髓间充质干细胞 转化生长因子Β 周期性拉伸应变 软骨细胞 分化 

分 类 号：R394.2[医药卫生—医学遗传学]