壳聚糖-脱细胞真皮三维材料作为骨组织工程支架材料的研究  

Study on the three-dimensional chitosan-decellularised-derma scaffolds for bone tissue engineering

作　　者：张燕[1] 李瑞欣[1] 张西正[1] 郭勇[1] 

机构地区：[1]军事医学科学院卫生装备研究所,天津300161

出　　处：《医用生物力学》2010年第1期11-15,共5页Journal of Medical Biomechanics

基　　金：国家自然基金重点项目(10832012)

摘　　要：目的观察MC3T3-El成骨前体细胞在壳聚糖-脱细胞真皮三维支架材料上的黏附情况,并评价其细胞相容性。方法通过冷冻干燥制备壳聚糖-脱细胞真皮三维支架材料,并测试其孔隙率、密度和吸水率,通过扫描电镜分析支架的微观形貌。采用体外培养细胞的方法,将MC3T3-E1细胞直接接种到壳聚糖-脱细胞真皮三维支架材料上,培养2,3,4,5h,各时间点各取3个样品,测定细胞在支架上的黏附率,确定最佳的细胞贴壁时间。将细胞接种到支架上,共培养1,3,5,7,9,11,13d,采用MTS方法绘制细胞增殖曲线,组织化学染色观察细胞形态,并利用材料试验机测试不同时间材料细胞复合物的压缩弹性模量。结果壳聚糖-脱细胞真皮材料具有连通的多孔结构,孔隙率为92.8%,密度为97.96g/L,吸水率为(2169±100)%。细胞相容性实验显示,成骨细胞易于在支架材料上黏附、增殖。结论壳聚糖-脱细胞真皮材料具有连通的孔隙,孔径较均匀,MC3T3-El成骨前体细胞易在壳聚糖-脱细胞真皮三维支架材料上黏附、增殖,表明该支架材料具有良好的细胞相容性。Objective To observe the adhesion of MC3T3-El osteoblastic progenitor cells to the three-dimensional chitosan-decellularised-derma scaffolds,and evaluate the cytocompatibility of the scaffolds.Method The three-dimensional chitosan-decellularised-derma scaffolds were prepared by the freeze-drying method,the porosity,density and water absorption of which were measured.The microscopic morphology of the composite scaffolds was analyzed by the scanning electron microscopy（SEM）.The MC3T3-E1 cells cultivated in vitro were seeded onto the composite scaffolds,and then co-cultured for 2,3,4 and 5 hours.At each time point,three specimens from each matrix were taken to determine the cell-adhesion rate and the best time of the cell-adhesion.The cells were seeded onto the composite scaffolds,and then co-cultured for 1,3,5,7,9,11 and 13 days.The MC3T3-E1 cells inside were evaluated with MTS test.The cell morphology was observed by the histological staining.The compression tests were performed using a Universal Testing Machine,at room temperature,as compared with no-cell-scaffolds.Results The three-dimensional chitosan-decellularised-derma scaffolds have high interval porosity with the porosity（92.8%）,the density（0.09796 g/ml）and the water absorption（2169±100）%.The cytocompatibility test shows that the seeded MC3T3-E1 cells can adhere to the scaffolds and proliferate.Conclusions The three-dimensional chitosan-decellularised-derma scaffolds have high interval porosity with the well-distributed diameter.The MC3T3-E1 cells are easy to adhere the scaffolds and proliferate which shows that the scaffolds have a good cytocompatibility.

关 键 词：壳聚糖 脱细胞真皮 空隙率 扫描电镜 骨组织工程 支架 

分 类 号：R318.01[医药卫生—生物医学工程]