抗凝血纳米壳聚糖微球的合成、表征及生物安全性评价  

Anticoagulant chitosan nanoparticles: synthesis,characterization and biological safety

作　　者：王子婧[1] 刘朝阳[1] 钱智勇[2] 高德伟[1] 郭希民[2] 

机构地区：[1]解放军总医院南楼综合外科,北京市100853 [2]解放军军事医学科学院,北京市100040

出　　处：《中国组织工程研究》2015年第47期7655-7659,共5页Chinese Journal of Tissue Engineering Research

摘　　要：背景:有研究显示,壳聚糖等天然多糖经磺化改性后具有类似肝素的抗凝功能,因磺酸化后的壳聚糖其形成的磺酸根基团与肝素的活性基团相似,具有良好的抗凝血性。目的:制备具有抗凝血功能的纳米壳聚糖微球,检测其形态结构、理化性能及生物安全性。方法:利用乳相法合成纳米壳聚糖微球,通过磺化反应合成磺酸化壳聚糖微球,通过透射电镜描述其形态特征,红外光谱观察其特异基团峰值变化。1凝血实验:分别将肝素、纳米壳聚糖微球及10,30,50 mg磺酸化壳聚糖微球加入SD大鼠血中,检测凝血指标。2溶血实验:分别将去离子水、生理盐水及10,30,50 g/L磺酸化壳聚糖微球浸提液加入兔2%红细胞悬液中,检测溶血率。3细胞毒性实验:分别采用含体积分数10%胎牛血清的DMEM培养基及10,30,50 g/L磺酸化壳聚糖微球的浸提液培养人脐静脉血管内皮细胞,72 h后检测细胞相对增殖率及毒性分级。结果与结论:扫描电镜显示磺酸化壳聚糖微球具有良好的形态结构,粒径大小50 nm,红外图谱提示存在磺化取代。体外凝血实验表明磺酸化壳聚糖微球具有显著抗凝血作用,抗凝血效果呈剂量效应关系。磺酸化壳聚糖微球符合国标关于溶血率小于5%的安全标准,无致溶血性。细胞毒性实验表明磺酸化壳聚糖微球浸提液无明显细胞毒性,其生物安全性符合国家标准。BACKGROUND：Studies have shown that chitosan and other natural polysaccharides have heparin-like anticoagulant function after sulfonated modification. Sulfonated chitosan has good anticoagulant property because the sulfonate group formed by sulfonated chitosan is similar with the active group of heparin. OBJECTIVE： To prepare the anticoagulant chitosan nanoparticles and to detect its morphology, physical and chemical properties and biological security. METHODS： Chitosan nanoparticles were synthesized by emulsion-chemical cross link. Sulfonated chitosan nanoparticles were synthesized by sulfonation reaction. Its morphology was described by transmission electron microscope. The peak-value change of its specific groups was observed by infrared spectroscopy. （1） Coagulation experiment： Heparin, chitosan nanoparticles and 10, 30 and 50 mg of sulfonated chitosan nanoparticles were added into the blood of Spraque-Dawley rats. The coagulation indicators were detected. （2） Hemolysis experiment： deionized water, physiological saline and 10, 30, 50 g/L sulfonated chitosan nanoparticles extracts were added into 2% red blood cel suspension of rabbits. The hemolysis rate was detected. （3） Cytotoxicity experiments： DMEM medium containing fetal bovine serum and 10, 30, 50 g/L sulfonated chitosan nanoparticle extracts were used to culture human umbilical vein endothelial cels. Cel relative growth rate and toxicity grading were detected after 72 hours. RESULTS AND CONCLUSION： Scanning electron microscopy showed that sulfonated chitosan nanoparticles had good morphology, with a diameter of 50 nm. Infrared spectroscopy showed that the sulfonated replacement occurred.In vitro coagulation experiments showed that sulfonated chitosan nanoparticles had significantanticoagulant effects in a dose-dependent manner. Sulfonated chitosan nanoparticles meet the national safety standard for hemolysis rate of less than 5%, non-induced hemolysis property. Cytotoxicity assays showed that sulfonated chitosan nanoparticles ex

关 键 词：生物材料 纳米材料 壳聚糖 微载体 磺酸化 抗凝血 生物安全性 

分 类 号：R318[医药卫生—生物医学工程]