壳聚糖与型胶原复合制作组织工程软骨支架及其性能研究  

FABRICATION AND PROPERTIES OF A COMPOSITE CHITOSAN/TYPE Ⅱ COLLAGEN SCAFFOLD FOR TISSUE ENGINEERING CARTILAGE

作　　者：史德海[1] 蔡道章[1] 周长忍[2] 李立华[2] 曾春[1] 李晓峰[1] 

机构地区：[1]中山大学附属第三医院骨科,广州510630 [2]暨南大学生命科学技术学院生物材料实验室

出　　处：《中国修复重建外科杂志》2005年第4期278-282,共5页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：广东省科技计划资助项目 (2 0 0 3A30 2 1 0 2)~~

摘　　要：目的　探讨采用壳聚糖与 型胶原复合制作新型组织工程软骨三维多孔支架的方法,并对其理化性能进行检测。　方法　将精制88%脱乙酰度壳聚糖溶于0 .2 mol/ L 醋酸溶液制成2 %溶液,高纯度猪 型胶原溶于0 .5 m ol/ L醋酸溶液制成1%溶液,两者按4∶1(重量比)充分搅拌混合,采用冷冻干燥法制成壳聚糖与 型胶原复合支架。采用碳化二亚胺/ N-羟基琥珀酰亚胺对支架进行交联,力学测定比较支架交联前后的强度变化,扫描电镜观察其超微结构,于2、4、6和8周经溶菌酶体外降解实验测定其体外降解性。　结果　制备的复合支架成形良好,交联后其力学强度明显增加。扫描电镜显示壳聚糖与 型胶原成分在支架内分布均匀,支架内孔洞相互连通似海绵状,孔径10 0～2 5 0 μm。各时间段复合支架体外降解较单纯壳聚糖支架快。　结论　壳聚糖与 型胶原复合成功地制作成了三维多孔复合支架。其理化性能及体外降解测定,可以作为支架载体,应用于组织工程软骨的构建。Objective To develop a novel porous three-dimensional scaffold and to investigate its physico-chemical properties for tissue engineering cartilage. Methods Refined 88% deacetylation degree chitosan was prepared and dissolved in 0.2 mol/L acetate acid and fully mixed with highly purified porcine type Ⅱcollagen in 0.5 mol/L acetate acid solution in a ratio of 4 to 1 (wt/wt). Freeze-drying process was employed to fabricate the composite scaffold. The construct was cross-linked by use of 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS). A mechanical tester was utilized to determine the tensile strength change before and after cross-linking. The microstructure was observed via scanning electron microscopy (SEM). The lysozyme degradation was performed to evaluate the degradability of the scaffold in vitro. Results A bulk scaffold with desired configuration was obtained. The mechanical test showed that the cross-linking treatment could enhance the mechanical strength of the scaffold. The SEM results revealed that the two constituents evenly distributed in the scaffold and that the matrix was porous, sponge-like with interconnected pore sizing 100-250 μm. In vitro lysozyme degradation indicated that cross-linked or uncross-linked composite scaffolds had faster degradation rate than the chitosan matrix. Conclusion Chitosan and typeⅡcollagen can be developed into a porous three-dimensional scaffold. The related physico-chemical tests suggest that the composite socaffold meets requirements for tissue engineered scaffold and may serve as an alternative cell-carrier for tissue engineering cartilage.

关 键 词：胶原复合 壳聚糖 性能研究 软骨支架 制作 N-羟基琥珀酰亚胺 组织工程软骨 mol/L 三维多孔支架 Ⅱ型胶原 复合支架 扫描电镜观察 体外降解 理化性能 碳化二亚胺 冷冻干燥法 脱乙酰度 强度变化 超微结构 支架成形 胶原成分 

分 类 号：R318.0[医药卫生—生物医学工程]