应用细胞膜片复合硅胶管内支撑构建组织工程管状软骨

Construction of Scaffold-Free Cylindrical Cartilage Combined with Cell Sheet and Silicon Tube

目的研究利用羊耳软骨细胞形成细胞膜片，构建无细胞支架组织工程管状软骨的可行性。方法体外培养扩增羊耳软骨细胞至第2代，以2．0×10^6cells／mL的高密度，培养7d，形成直径为35mm的圆形细胞膜片，将细胞膜片均匀包裹于硅胶管，植入裸鼠背部皮下，4周后取材检测。以大体观察、组织学、免疫组织化学等方法，对形成的软骨进行评价。结果大体观察可见形成良好的管状软骨，HE染色见软骨陷窝形态规则，番红0染色及Ⅱ型胶原免疫组化均呈阳性表达。结论细胞膜片复合硅胶管内支撑的方法能形成管状软骨，为气管软骨的再造提供了新的方法。

Objective To explore the feasibility of fabricating a scaffold-free cylindrical cartilage using cell sheet with silicon tube as interior support in vivo. Methods Auricular chondrocytes were harvested from goat and the second passage of auricular chondrocytes were cultivated with high-density of 2.0~106 cells/mL to form a chondrocyte sheet with the diameter of 35 ram. Then the silicon tube covered with cell sheet was implanted into the back of nude mice. The tissue engineered cylindrical cartilages were evaluated by gross observation, histological and immunohistochemical examination. Results The cartilage was sufficiently elastic and stiff to maintain the structure without disruption. Cartilage tissue was observed by HE staining, Safranin O staining and immunohistochemical staining for type ]1 collagen. Conclusion A novel technique was developed for fabricating engineered cylindrical cartilage using cell sheet with silicon tube in vivo.