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
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国家自然科学基金资助项目(10832012)


Study on the three-dimensional chitosan-decellularised-derma scaffolds for bone tissue engineering
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    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、5 h,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、13 d,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。

    Abstract:

    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. Methods: 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, in order to ascertain 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 had high interval porosity with the porosity (92.8%), the density (0.09796g/ml) and the water absorption (2169±100)%. The cytocompatibility test showed that the seeded MC3T3-E1 cells could adhere to the scaffolds and proliferate.Conclusion:The three-dimensional chitosan-decellularised-derma scaffolds had high interval porosity with the well-distributed diameter. The MC3T3-E1 cells were easy to adhere the scaffolds and proliferate which showed that the scaffolds had a good cytocompatibility.

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张燕,李瑞欣,张西正,郭勇.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[J].医用生物力学,2010,25(1):11-15

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