β-连环蛋白对猪关节软骨细胞合成糖胺多糖的影响

Effects of β-catenin on glycosaminoglycan in pig articular chondrocytesl

目的探讨β-连环蛋白对软骨细胞合成糖胺多糖含量的影响。方法通过Ⅱ型胶原酶消化法获得高活性的软骨细胞，定量接种6×10^6个细胞于培养皿，在细胞贴壁后更换培养基时，加入β-连环蛋白10μg/L，每周传代一次，连续5周，留取所有培养液，用阿利新蓝法测定软骨细胞糖胺多糖的变化；利用光镜观察原代及传代软骨细胞的生长情况。以50×10^6／ml浓度均匀接种于经聚乳酸包埋聚羟基乙酸（Polyglycolic acid，PGA）高分子聚合物支架，形成细胞支架复合体，扫描电镜观察细胞生长情况。结果体外培养的软骨细胞生长情况良好；软骨细胞从传代后合成糖胺多糖类基质，第2代传代细胞分泌GAG的能力最强；加入β-连环蛋白能明显促进传代软骨细胞合成大量的糖胺多糖类基质；扫描电镜显示软骨细胞与材料具有良好的相容性，培养7d有基质沉积。结论β-连环蛋白能明显促进体外培养的传代软骨细胞合成大量的糖胺多糖类基质；PLA包埋PGA是软骨细胞良好的支架。

Objective To explore the effect of β- catenin on synthesis of glycosaminoglycan in pig articular chondrocytesl. Methods The highly active chondrocytes were isolated by Ⅱ collagenase digestion and 6 × 10^6 cells were cultured in Ham, β-catenin with 10 μg/L were put in the medium for up to 5 weeks. Alcian blue chromatomery was used to detect the change of glycosaminoglycan （GAG） in articular ehondrocytes. The growth of origin and passages cartilage cells were observed by light microscope. About 50 ×10^6/ml cells were seeded onto PGA high molecular polymer scaffoht coated with polylactic acid （PLA） to form cell-scaffold complex which was observed by scanning electron microscope （ SEM ） observation. Results The growth of second chondrocytes cultured in virto was good; the chondrocytes treated by β-catenin synthesized more GAG from 2 to 5 weeks. The good compatibility with chondrocytes to PGA was observed through SEM. Histologiccally, constructs consisted of chondrocytes embedded in homogeneous cartilaginous matris. Conclusion β-catenin could stimulate the synthesis of the GAG by cultured chondrocytes. PGA scaffold coated with PLA is a good polymer.