壳聚糖电介质复合物作为软骨再生支架实验研究

Experimental Study on Chitosan-Polyelectrolyte Complex as a Scaffold for Cartilage Regeneration

目的探讨壳聚糖电介质复合物对髁突软骨细胞增殖、吸附等功能的影响.方法合成磷酸化壳聚糖电介质复合物(CS-PEC)制备三维凝胶支架,并与髁突软骨细胞体外培养,光镜和扫描电镜及MTT等方法检测其对软骨细胞增殖、吸附的影响.结果壳聚糖(chitosan,CS)、CS-PEC支架接种细胞悬液后,细胞形态、铺展良好.在CS-PEC支架中,48～72 h内示细胞形态、铺展良好,逐渐通过伪足相互联系,证明此支架的性能优于单纯CS支架.MTT检测结果显示培养至第5天时,CS-PEC组细胞增殖活性强于CS组,两组之间细胞增殖活性有显著性差异(P＜0.05).结论软骨细胞在CS-PEC支架中的吸附及增殖状态良好,此支架有可能作为软骨再生的载体.

Objective To investigate whether Chitosan-Polyelectrolyte Complex （CS-PEC） is appropriate scaffold for ehondroeyte culture of cartilage regeneration. Methods Condylar chondrocytes of the fetal mouse were seeded onto the three-dimension gels scaffolds of CS-PEC for culture. Adhesion of the ehondrocytes in scaffolds was observed by light microscope and scanning electron microscope. The function of chondrocytes in scaffolds was detected by MTr automated colormetric microaasay. Results The majority of cells attached to the CS, CS-PEC surface assumed a fibroblast-like morphology, and the cell population expanded rapidly. After 48-72 h, chondrocytes attached to the CS-PEC largely maintained a round or polygonal morphology. After 3 days culture, the cells became confluent on the scaffold and acquired a predominantly spherical or sometimes fusiform shape. It would take about one week for cells cultured in the CS scaffolds. MTF results showed cell proliferation between the CS and CS-PEC groups are significantly different （ P 〈 0.05） for culture 5 days. Conclusion Porous CS-PEC scaffold may be a more suitable scaffold than CS for chondrocytes culture of cartilage regeneration.