软骨细胞与复合水凝胶的生物相容性研究

Study of biocompatibility of chondrocytes and composite hydrogel

目的构建光固化壳聚糖与自固化海藻酸钠复合水凝胶体系,研究其与软骨细胞的体外生物相容性,探索其作为软骨组织工程支架的可行性。方法以三维多孔光固化壳聚糖水凝胶作为细胞的空间载体,利用自固化海藻酸钠水凝胶将软骨细胞胶封入光固化壳聚糖水凝胶支架,构成复合水凝胶-细胞复合体。采用扫描电子显微镜观察光固化壳聚糖的结构形貌以及细胞在支架内部的分布与形态;CCK8法绘制细胞增殖曲线;DAPI细胞核染色荧光显微镜观察细胞核形态变化,判断细胞活性。结果纯光固化壳聚糖水凝胶材料呈孔隙较为均匀一致的多孔结构,平均孔径为300μm左右。复合水凝胶能够模拟天然软骨细胞外基质环境,使细胞保持圆形生长状态;促进成软骨细胞的增殖,具有良好的生物相容性。结论复合水凝胶能够为软骨细胞提供一个类似天然的生活环境,并能够促进其增殖、生长,有望作为软骨组织工程支架应用于软骨缺损修复的实验研究。

Objective To evaluate biocompatibility of chondrocytes with composite hydrogel system of light-cured chitosan and sodium alginate, and to study its feasibility for cartilage tissue engineering. Methods Three-dimensional porous light- cured chitosan hydrogel was prepared as the carrier of chondrocytes in space. Chondrocytes were enclosed into light-cured chitosan hydrogel by means of sodium alginate hydrogel. The structure and morphology of light-cured chitosan scaffolds and cell distribution and morphology within scaffolds were observed by scanning electron microscopy （ SEM）. The method of CCK8 was used to draw the cell growth curve and cell activity was tested by DAPI staining. Results Pure light curing chitosan hydrogel material presented as a porous structure of uniform pores with the average pore size about 300 p^m. Composite hydrogel could mimic natural cartilage extracellular matrix environment to maintain the chondroeytes as natural round state and could promote the proliferation of chondrocytes. Conclusion Composite hydrogel of light-cured chitosan and sodium alginate can provide a similar natural biological environment for chondrocytes, and promote their proliferation and growth, which may be a very promising cartilage tissue engineering scaffolds for in vivo experimental studies of cartilage defects.