羟丁基壳聚糖的制备及其与鼠胚胎成纤维细胞的生物相容性研究

Preparation of Hydroxybutyl Chitosan and Its Biocompatibility Evaluation with Mouse Embryonic Fibroblasts

目的制备温敏型壳聚糖衍生物羟丁基壳聚糖(Hydroxybutyl chitosan,HBC),研究鼠胚胎成纤维细胞(Mouseembryonic fibroblasts,MEF)与HBC的相容性,探讨羟丁基壳聚糖作为组织工程材料的可行性。方法对壳聚糖化学改性制备HBC,研究其最佳制备条件。使用不同溶剂对HBC进行溶解实验;利用红外光谱、扫描电镜对产物结构进行观察。从小鼠胚胎中分离培养MEF;MTT法测定HBC对MEF活性的影响。结果制备的4种HBC均能溶于水,55℃反应14 h的HBC 4水溶性较好。HBC溶液在37℃具有明显的溶胶-凝胶的转变,通过物理交联形成凝胶并可逆。不同反应条件下制得的4种HBC浸提液对MEF细胞的毒性均在0级或1级,表现出良好的生物相容性。结论成功制备羟HBC,HBC具有良好的温度敏感性及体温(37℃)下凝胶的特性,与MEF生物相容性良好,是一种具有应用前景的新型组织工程材料。

Objective To prepare and study the biocompatibility of Hydroxybutyl chitosan（HBC） with mouse embryonic fibroblasts（MEF） in vitro.To investigate the feasibility of using HBC as tissue engineering materials.Methods The HBC were prepared by chitosan chemically modified and the best preparation conditions were summarized.Different solvents were used on HBC for the solubility test.The structure of the HBC was characterized by IR and scanning electron microscopy（SEM）.MEFs were isolated and cultivated from mouse embryo.The biocompatibility of HBC is investigated by using the MTT assay.Results All the four HBC samples could dissolve in water and HBC 4 which was acted 14 hours at 55 ℃ have better water-solubility.HBC solution can rapidly transform to semisolid at body temperature（37 ℃） and revert to its solvated state by physical cross-liking at lower temperature.MEF cultured with four kinds of leaching of HBC were investigated by the MTT assay and the cytotoxicity of HBC was in graded 0 and 1.Conclusion HBC was prepared successfully with good thermo-sensitivity and could be transformed into gelation at body temperature（37 ℃）.HBC presented good biocompatibility with MEFs and could be used as tissue engineering materials.