兔软骨细胞在可注射温敏型壳聚糖／聚乙烯醇凝胶中生长的研究

The study of rabbit chondrocytes growth in injective chitosan/polyvinyl alcohol gel composite

目的探讨以壳聚糖（CS）和聚乙烯醇（PVA）制备的可注射温敏型凝胶作为兔软骨细胞生长支架的可行性。方法将壳聚糖和聚乙烯醇溶液混合制成温敏型凝胶，取第三代软骨细胞接种于凝胶支架。于接种后24、48和72h采用MTT测定细胞活性及毒力；于1、2及3周后采用扫描电镜及共聚焦激光扫描荧光显微镜观察软骨细胞在凝胶中的形态及生长状态。结果MTT结果显示接种细胞数量随着接种细胞生长时间的推移而明显增加，各组之间差异有统计学意义（P〈0．05）。扫描电镜及激光共聚焦荧光显微镜观察表明软骨细胞在CS／PVA凝胶支架中生长良好。软骨细胞在凝胶支架中保持了很高的增殖能力，材料对细胞无明显不良反应。结论壳聚糖／聚乙烯醇混合凝胶可作为兔软骨细胞培养的生长支架，应用于软骨组织工程。

Objective To investigate the composite of chitosan（CS） and polyvinyl alcohol （PVA） as scaffold carrier for rabbit chondrocytes nurture and growth. Methods The third passage of chondrocytes were seeded in CS/PVA gel scaffold and 24, 48 and 72 h after which cytoactive and toxicity were determined by MTT respectively. After one, two and three weeks, the growing status and morphology of chondrocytes in CS/PVA gel were observed with scaning electron microscope （SEM） and laser confocal scanning fluorescence microscope （LCSM）. Results The third passage of chondrocytes in CS/PVA gel scaffold remained high proliferation ability. MTT measuring cell activity and virulence, the result showed that the number of ceils obviously increased with the time, with statistical significance of difference between each groups （P 〈 0.05）, without side effect to cells by the material. Observation of scaning electron microscope and confocal laser scanning fluorescence microscope showed that chondrocytes grew well with the scaffold of CS/ PVA gel. Conclusion CS/PVA mixed gel material can be used as scaffold for rabbit chondrocytes growing for repairing cartilages defect in tissue engineering.