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目的 探讨以藻酸盐为支架材料,经体外软骨向诱导的大鼠骨髓间充质干细胞为种子细胞,在体内异位 形成组织工程化软骨的可能性。方法 32只雄性SD大鼠随机平均分为实验组和对照组两组。实验组分离培养自 体骨髓间充质干细胞,传至第3代经转化生长因子β等诱导因子诱导10d后与藻酸钠复合,并滴加氯化钙使其成 凝胶状后注入大鼠背部皮下。对照组只注入藻酸盐。术后4周、8周取材进行苏木精-伊红、奥新蓝染色及透射电 镜观察。结果 实验组术后8周肉眼可见软骨形成,苏木精-伊红、奥新蓝染色发现有大量软骨形成,软骨内可见 细胞团,软骨细胞周围基质丰富,藻酸盐降解明显,未降解的藻酸盐松散的分布于软骨间。对照组未见软骨样组织 形成。结论 经软骨向诱导的自体骨髓间充质干细胞结合藻酸盐凝胶可在体内异位形成较理想的组织工程化软 骨。
OBJECTIVE: To investigate the possibility of ectopic formation of tissue-engineered cartilage in vivo using alginate-induced rat bone marrow-derived mesenchymal stem cells as seed cells. Methods Thirty-two male Sprague-Dawley rats were randomly divided into experimental group and control group. The experimental group were cultured with autologous bone marrow mesenchymal stem cells, which were induced to pass into the third generation induced by induction factor such as transforming growth factor β and combined with sodium alginate 10 days later. After calcium chloride was added into the experimental group, they were injected into the back of rats Subcutaneous. The control group was infused with alginate only. At 4 weeks and 8 weeks after operation, hematoxylin and eosin were stained and observed with transmission electron microscopy. RESULTS: Cartilage formation was observed 8 weeks after operation in the experimental group. Massive cartilage was found in hematoxylin-eosin and Azoin-blue staining, and cell mass was found in the cartilage. Enriched matrix around the chondrocytes, obvious degradation of alginate, undegraded Alginate loosely distributed between the cartilage. No cartilage-like tissue was found in the control group. Conclusion The combination of alginate gel induced by autologous bone marrow mesenchymal stem cells induced by cartilage can form more ideal tissue-engineered cartilage in vivo.