骨髓间充质干细胞复合温敏水凝胶支架的制备

摘要

背景：壳聚糖及其衍生物制备的支架对细胞迁移和神经轴突再生有重要作用。壳聚糖及其衍生物的组织相容性好，易使干细胞在其表面附着生长，在神经组织工程具有较为广阔的应用前景。　　目的：制备适宜骨髓间充质干细胞生长的壳聚糖/壳聚糖季铵盐/甘油磷酸钠温敏性水凝胶细胞支架，观察骨髓间充质干细胞在细胞支架中的生长情况。　　方法：将壳聚糖进行季铵盐化改性处理，通过傅里叶变换红外光谱分析谱检测确定其生成。实验以壳聚糖与壳聚糖季铵盐配比为8∶1成功制备出较为稳定的壳聚糖/壳聚糖季铵盐/甘油磷酸钠温敏性温敏水凝胶细胞支架，观察成胶情况，并进行生物安全性检测。　　结果与结论：实验在傅里叶变换红外光图谱上发现了季铵基基团的特征峰。细胞毒性实验显示，水凝胶浸提液干预的大鼠骨髓间充质干细胞无毒性。急性全身毒性实验显示，浸提液对大鼠体质量增加无明显影响，支架生物安全性较好。扫描电镜观察显示，骨髓间充质干细胞在细胞支架中能正常的生长和增殖。结果证实，实验成功制备了壳聚糖/壳聚糖季铵盐/甘油磷酸钠温敏性水凝胶细胞支架，适合骨髓间充质干细胞生长和增殖。%BACKGROUND:Scaffolds made of chitosan and its derivatives play an important role in cellmigration and axonal regeneration. Chitosan and its derivatives have good histocompatibility, which is easy to make stem cells grow on the surface, thereby having a more broad application prospect in the nerve tissue engineering. OBJECTIVE:To fabricate a thermosensitive hydrogel scaffold using chitosan/hydroxypropyltrimethyl ammonium chloride chitosan/sodium glycerophosphate (CS/HACC/GP), which is suitable for cellgrowth, and then, to observe the growth and proliferation of bone marrow mesenchymal stem cells on the scaffold. METHODS:Chitosan was modified using quaternary ammonium salt and confirmed by Fourier transform infrared spectroscopy. The chitosan and quaternary ammonium salt of chitosan was mixed at a ratio of 8:1 to successful y prepare stable CS/HACC/GP thermosensitive hydrogel scaffold. Then, the gel ing was observed, and biosafety test was conducted. RESULTS AND CONCLUSION:Fourier transform infrared spectroscopy showed the characteristic peak of quaternary ammonium groups. Cytotoxicity test showed that rat bone marrow mesenchymal stem cells cultured in hydrogel extracts had no toxicity. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide test showed that hydrogel extracts exerted no significant effect on the increase in body weight, and the biological safety of the scaffold was good. Under the scanning electron microscopy, bone marrow-derived mesenchymal stem cells grew and proliferated normal y in the scaffold. The results confirmed that the CS/HACC/GP thermosensitive hydrogel scaffold was successful y prepared in the experiment, which is suitable for the growth and proliferation of bone marrow mesenchymal stem cells.