CAJ | 학술논문

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。
배경：연구표명전화생장인자β대골수간충질간세포적연골방향분화구유현저적유도작용。주기성랍신응변가이모의연골세포재체내적역학배경，대세포적증식화분화기착중요적조절작용。목적：탐토전화생장인자β급주기성랍신응변재유도골수간충질간세포향연골양세포분화과정중시부구유협동작용。방법：취2월령신서란대백토10지，골천침천입고골수강내，추취골수3.0-4.0 mL병분리배양골수간충질간세포，전지3대후수궤분위4조，분별위공백조、전화생장인자β조、주기성랍신응변조이급주기성랍신응변+전화생장인자β조，작용1，3，6 d후취출상응세포，번홍O염색관찰대체형태，아이신람염색검측당알취당수평，ELISA검측상청액기질금속단백매13급기질금속단백매조직억제제1수평，RT-PCR검측Ⅱ형효원、기질금속단백매13급기질금속단백매조직억제제1 mRNA상대표체량。결과여결론：번홍O염색가견세포정장사형혹불규칙삼각형양개변，각실험조교공백조세포수량급기질분비증다。작용제3천시전화생장인자β조、주기성랍신응변+전화생장인자β조상청액당알취당수평교공백조균승고(P <0.05)，주기성랍신응변+전화생장인자β조Ⅱ형효원mRNA상대표체량교공백조증고(P <0.05)。결과제시전화생장인자β급주기성랍신응변균가유도골수간충질간세포향연골세포분화，이자구유명현적협동작용。
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 differentiation of bone marrow mesenchymal stem cells into chondrocytes, showing an apparent cooperative action.