壳聚糖/磷酸化壳聚糖纳米复合物的制备及细胞相容性研究

Fabrication and Cytocompatibility of the Nanocomposite Chitosan and Phosphorylatd Chitosan

目的:由壳聚糖(chitosan,CS)和磷酸化壳聚糖(phosphorylated chitosan,PCS)通过静电自组装仿生合成一种新型骨修复材料并体外评价其细胞相容性。方法:对CS进行磷酸根接枝,合成PCS,然后将PCS溶液逐滴滴加到CS的醋酸溶液中,得到壳聚糖/磷酸化壳聚糖复合物(CS/PCS)。将CS/PCS水胶体置于饱和的Ca(OH)_2溶液中矿化供细胞培养用。第3～5代成骨细胞与矿化后的CS/PCS复合物共同培养,以羟基磷灰石组为对照组。倒置显微镜下观察细胞的黏附、生长情况。MTT法检测细胞的增殖;碱性磷酸酶(alkaline phosphatase,ALP)检测细胞的分化。结果:CS/PCS复合物呈多孔网状结构,矿化后的CS/PCS可以促进成骨细胞的增殖和分化。结论:仿生合成的CS/PCS复合水胶体矿化后显示了良好的细胞相容性,可以促进成骨细胞的增殖和分化,可作为一种新型的骨修复材料。

Objective： To synthesize a new bone repair material composed of chitosan and phosphorylated chitosan （CS/PCS） and to evaluate its cytocompatibility in vitro. Methods： Phosphate group was incorporated into chitosan （CS） to form phosphorylated chitosan （PCS）. The PCS solution was dropped into the acetic acid solution of CS to get the complex of chitosan and phosphorylated ehitosan （CS/PCS）. CS/PCS hydroeolloid was co-cultured with rat osteoblasts in vitro to observe the adhesion and growth of osteoblasts. Hydroxyapatite group was used as control. The proliferation and differentiation of osteoblasts were determined by MTT and alkaline phosphatase methods. Results： The structure of CS/ PCS composite was porous and reticular. It was evident that CS/PCS could promote osteoblasts proliferation and differentiation. Conclusion： The biomimetic CS/PCS composite has well cytocompatibility with osteoblasts after mineralizing in saturated Ca（OH）2 solution. CS/PCS composite can promote proliferation and differentiation of osteoblasts.