体外重建组织工程关节软骨的实验研究

Constructing tissue engineered articular cartilage in vitro

目的 用胶原蛋白和人血纤维蛋白混合物为载体在体外进行软骨细胞三维立体培养 ,构建人工软骨组织。方法 取 2周龄的新生兔关节软骨 ,经消化 ,将获得的软骨细胞与牛 型胶原、人血冻干纤维蛋白原、凝血酶按一定比例混合 ,制成软骨培养物并在体外培养。培养第 3周时 ,取材进行 HE、甲苯胺蓝染色和透射电镜检查。结果 体外培养 3周 ,培养物内细胞均存活 ,形成软骨陷窝 ,同源性细胞簇出现 ,并分泌软骨基质。透射电镜下可见丰富的粗面内质网和线粒体 ,及少量的高尔基复合体。结论 用胶原蛋白和人血纤维蛋白为载体支架体外培养软骨细胞 。

Objective To construct articular cartilage by three dimensional culturing chondrocytes using collagen and human fibrin as supporter.Methods Chondrocytes were isolated from articular cartilage of 2 week old rabbit,digested by collagenase II, and mixed with bovine I type of collagen, human blood freeze dry fibrinogen and thrombosin according to a certain proportion to culture in vitro.Results After 3 weeks, all cells were viable,formed cartilage lacuna, appeared homologous cell cluster,and secreted cartilage matrix stained by toluidine blue in culture. Abundant rough endoplasmic reticulum and mitochondria, as well as a small amount of golgl complex were oberserved under transmission electron microscope.Conclusions The biggish artificial cartilage tissue can be constructed by three dimensional culturing hondrocytes using collagen and human fibrin as supporter.