抗凝血纳米壳聚糖微球的合成、表征及生物安全性评价

发布时间：2018-07-11 14:00

本文选题：壳聚糖 + 纳米结构　；参考：《中国组织工程研究》2015年47期

【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。
[Abstract]:Background: studies have shown that chitosan and other natural polysaccharides modified by sulfonation have heparin-like anticoagulant function, because the sulfonic acid group formed by the sulfated chitosan is similar to the active group of heparin, so it has good anticoagulant effect. Aim: to prepare nano-chitosan microspheres with anticoagulant function, and to detect their morphology, physical and chemical properties and biological safety. Methods: Nano-chitosan microspheres were synthesized by emulsion method and sulfonated chitosan microspheres were synthesized by sulfonation reaction. The morphologic characteristics of chitosan sulfonated microspheres were described by transmission electron microscope. The peak changes of specific groups were observed by IR spectra. Nano-chitosan microspheres and 1030g / L sulfated chitosan microspheres were added to SD rats' blood. The blood coagulation index (.2) hemolysis test was performed: deionized water, normal saline and 1030g / L sulfonated chitosan microspheres were added to 2% red blood cell suspension of rabbits, respectively. The cytotoxicity of human umbilical vein endothelial cells (HUVECs) was determined by DMEM medium containing 10% fetal bovine serum (FBS) and extraction solution of 1030g / L sulfonic chitosan microspheres for 72 h. Results and conclusion: scanning electron microscope showed that the sulfonated chitosan microspheres had a good morphology, and the particle size was 50 nm. The infrared spectra indicated the existence of sulfonation substitution. In vitro clotting test showed that the sulfonic chitosan microspheres had a significant anticoagulant effect, and the anticoagulant effect was in a dose-dependent manner. The sulfonic chitosan microspheres meet the safety standard of hemolysis rate less than 5%, and no hemolysis. The cytotoxicity test showed that the extractant of sulfonic chitosan microspheres had no obvious cytotoxicity and its biological safety was in line with the national standard.
【作者单位】：解放军总医院南楼综合外科;解放军军事医学科学院;
【分类号】：R318.08

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