壳聚糖-胶原支架的制备与生物相容性研究

Research on preparation and biocompatibility of chitosan-collagen scaffold

目的将壳聚糖(Chitosan,CS)与胶原(Collagen,Col)混合制备骨软骨组织工程材料,并检测其物理性能和生物安全性,以期为骨软骨损伤修复提供一种生物支架材料。方法采用真空冷冻干燥法将CS和Col按不同比例制成三组混合支架,检测支架的孔隙率、吸水膨胀率、热水溶失率及力学性能,通过扫描电镜观察支架结构,采用CCK-8法、Factin染色法、Live/Dead细胞染色法检测其细胞毒性及生物相容性。结果支架为白色而规则的圆柱体,三组支架均具有良好的孔隙率、吸水膨胀率、热水溶失率及力学性能,扫描电镜显示三组支架均具有良好的多孔网格结构,其中当CS∶Col为1∶3时,物理性能最佳。CCK-8法检测结果显示,在一定的支架浸提时间内,与对照组相比,三组均未出现明显的生长抑制表现(P>0.05)。F-actin染色观察结果显示,各组细胞形态大小均一,细胞骨架形态规则,细胞核无异染及破碎现象,并且DAPI染色后发现细胞在支架内生长分布良好。Live/Dead细胞检测发现,三组支架均无细胞毒性,并且三组支架的细胞活率无明显差异(P>0.05)。结论CS-Col复合支架具有良好的生物相容性和物理性能,是一种有潜力的骨软骨组织工程材料。

Objective To prepare osteochondral tissue engineering materials by mixing chitosan(CS)and collagen(Col),and their physical properties and biological safety were tested,so as to provide a biological scaffold material for the repair of osteochondral injury.Methods The three groups of mixed scaffolds of CS and Col were made by vacuum freezedrying method in different proportions.The porosity,water absorption expansion rate,hot water dissolution rate and mechanical properties of the scaffolds were detected.The scaffold structure was observed by scanning electron microscope,and its cytotoxicity and biocompatibility were detected by CCK-8 method,F-actin staining method,and Live/Read cell staining method.Results The scaffolds were white and regular cylinders,and all three groups of scaffolds had good porosity,water absorption expansion rate,hot water dissolution rate and mechanical properties.Scanning electron microscopy showed that all three groups of scaffolds had good porous grid structure,and the physical properties were the best when CS︰Col was 1︰3.The results of CCK-8 method showed that,compared with the control group,there was no obvious growth inhibition in the three groups within a certain time(P>0.05).The results of F-actin staining showed that the cells in each group were uniform in size,the cytoskeleton was regular in shape,and the nucleus was not stained and broken.After DAPI staining,it was found that the cells grew and distributed well in the scaffold.Live/Read cell testing revealed that all three groups of scaffolds had no cytotoxicity to cells,and there was no significant difference in cell viability among the three groups of scaffolds(P>0.05).Conclusion CS-Col composite scaffold has good biocompatibility and physical properties,and it is a potential tissue engineering material for osteochondrosis.