Ⅱ型胶原-硫酸软骨素支架的制备及组织工程软骨的构建

Preparation of type Ⅱ collagen-CS scaffolds and application in tissue engineering cartilage

研究经乙基-(3-二甲基氨基丙基)碳化二亚胺盐酸盐(EDC)处理的Ⅱ型胶原-硫酸软骨素支架材料的性能特点,并在体外构建组织工程软骨。从鸡软骨中提取Ⅱ型胶原,以不同浓度的EDC为交联剂通过冷冻干燥的方法制备Ⅱ型胶原与硫酸软骨素复合支架并测定其理化性质。将体外培养的新生兔关节软骨细胞接种在Ⅱ型胶原与硫酸软骨素复合支架上,观察软骨细胞在支架上的生长形态并检测支架上软骨细胞分泌的糖胺聚糖含量及Ⅱ型胶原含量。结果表明:采用EDC与硫酸软骨素交联增加了支架的稳定性,最适的交联剂质量浓度为7 mg/mL。软骨细胞在复合支架上增殖分化良好,并保持软骨细胞特异分化的表型,分泌Ⅱ型胶原与蛋白多糖(GAG)。培养14 d后已有软骨样组织形成。

The preparation and characteration of type Ⅱ collagen-chondroitin sulfate（CS） scaffolds and the application in tissue engineering cartilage were studied.Porous type Ⅱcollagen-CS scaffolds were prepared using variable concentration of 1-ethyl-3（3-dimethyl aminopropyl） carbodiimide（EDC）.The physico-chemical properties of the collagen scaffolds were evaluated.Then,isolated chondrocytes from the rabbit articular cartilage were cultured in porous type Ⅱ collagen scaffolds in the presence of covalently attached CS up to 14 d.Cell proliferation,the total amount of proteoglycans and type Ⅱcollagen retained in the scaffold and chondrocytes morphology were evaluated.Results suggested that EDC-crosslinking was proved to be mechanically stable for scaffolds with increasing EDC concentration.On concentration of EDC higher than 7 mg/mL,the crosslinking stability was obtained.Cell proliferation and the total amount of proteoglycans and type Ⅱ collagen retained in the scaffolds were higher in type Ⅱ collagen-CS scaffolds.Scanning electron microscopy revealed a chondrocyte distributed throughout the porous surface of scaffolds with ellipse.