碱性成纤维细胞生长因子与胶原对组织工程软骨体外构建的影响

目的：以三维成团培养为培养系统，探讨bFGF与胶原对组织工程软骨体外构建的影响。方法：成团培养兔生长板软骨细胞，设bFGF、胶原及联合作用组。HE染色观察新生组织形态；免疫组化检测Ⅰ、Ⅱ型胶原表达以观察细胞表型；Hoechst 33258法检测细胞DNA含量；羟脯氨酸法与阿新蓝法测定基质中胶原与蛋白多糖的合成。结果：新生软骨的组织学形态近似自然软骨；各实验组软骨细胞DNA含量明显上升；胶原可以显著促进基质的合成；各实验组Ⅰ型胶原的表达少于对照组，Ⅱ型胶原的表达则高于对照组；联合作用组效果更加明显。结论：三维的成团培养可以促进基质合成，有效维持软骨细胞表型；bFGF与胶原有利于工程化软骨构建，其效果具有协同效应，两者联合应用可进一步促进软骨再生。

Effects of bFGF and collagen on the construction of tissue-engineered cartilage in vitro

Objective: To investigate effects of bFGF and collagen on tissue-engineered cartilage in pellet culture. Methods: Growth-plate chondrocytes were incubated in pellet culture. bFGF and Collagen was applied alone or together based on the results of previous research.The histological structure of neocartilage was observed by stained with H＆E and Immunohistochemical analysis. The DNA quantity of neocartilage was measured by hoechst33258 dye.The production of matrix was estimated based on hydroxyproline quantity and Alcian Blue stain method. Results: The histological structure of in vitro-formed tissue was similar to growth-plate cartilage.The DNA quantity of neocartilage increased by the supplement of bFGF and collagen. The production of matrix was promoted by collagen. bFGF and collagen suppressed the expression of collagen Ⅰ,and decelerated the decrease of collagenⅡ. The combined usage of bFGF and collagen had synergistic effects. Conclusion: Pellet culture maintained the phenotype of chondrocyte and promoted matrix production. bFGF and Collagen improved the construction of engineered cartilage. The combined usage of bFGF and collagen had synergistic effects on the regeneration of cartilage.