【摘要】 背景:采用组织工程技术再生和重建软骨是目前修复软骨组织缺损效果最好、最有应用前景的方法。目的:以体外培养的软骨细胞和交联透明质酸钠为支架材料,开发一套体外构建组织工程软骨的完整方案。方法:分离新西兰兔膝关节软骨细胞,制成细胞悬液滴加于交联透明质酸钠支架上,体外复合培养21 d,提取RNA进行RT-PCR检测,制备冰冻切片进行显微观察和免疫组织化学观察。结果与结论:软骨细胞接种于交联透明质酸钠支架材料后,可贴附于支架上生长,并且大量细胞聚集成团,在支架材料的纤维间隙中生长或呈单层细胞附着于支架材料纤维。细胞-支架复合物表达软骨组织特异性蛋白聚糖基因和Ⅱ型胶原α1基因,以及软骨组织特异性蛋白Ⅱ型胶原蛋白,可维持软骨细胞表型。表明培养的细胞-支架复合物在体外培养可形成软骨细胞外基质,有望获得组织工程软骨组织。更多还原

【Abstract】 BACKGROUND: Tissue engineering provides new ideas and approaches for repair of cartilage defects. OBJECTIVE: To develop a complete set of solutions for construction of tissue engineered cartilage in vitro,with chondrocytes as seed cells and cross-linked sodium hyaluronate as scaffold materials. METHODS: New Zealand rabbit articular chondrocytes were isolated,counted,and then cultured and passaged to prepare cell suspension. Toluidine blue staining,RT-PCR and immunocytochemistry were exerted to evaluate the cultured cells. Chondrocytes were seeded and co-cultured with cross-linked sodium hyaluronate scaffold for 21 days. Then,RNA was isolated for RT-PCR,and frozen sections were prepared for morphological observation and immunohistochemistry study. RESULTS AND CONCLUSION: The chondrocytes could adhere to the cross-linked sodium hyaluronate scaffold and aggregate,growing between fibers or adhering to the scaffold in a monolayer manner. The transcripts of cartilage specific aggrecan gene and collagen type II alpha 1 gene and cartilage specific protein collagen type II were expressed in cell-scaffold complexes to maintain the phenotype of chondrocytes. Cell-scaffold complexes co-cultured in vitro can form cartilage extracellular matrix,by which tissue engineered cartilage is expected to be obtained.更多还原