载阿霉素新型壳聚糖纳米粒的性质及体外抗肿瘤研究

Characterization and the anti-tumor effect of a novel doxorubicin chitosan nanoparticle in vitro

目的采用离子凝胶法与化学交联法联合制备新型壳聚糖纳米粒,考察其相关性质以评估此种壳聚糖纳米粒的潜在的应用价值。方法以盐酸阿霉素为模型药物,采用离子凝胶法和化学交联法联合制备新型壳聚糖纳米粒。以离子凝胶法制备的壳聚糖纳米粒为对照,离心法测定纳米粒的包封率;膜透析法检测纳米粒在不同pH下的盐酸阿霉素累积释放度;考察纳米粒在不同介质及pH中的粒径、电位和多分散系数。以离子凝胶法制备的壳聚糖纳米粒及游离盐酸阿霉素为对照组,MTT法检测新型壳聚糖纳米粒对人乳腺癌MCF-7肿瘤细胞的体外抑制作用。结果新型壳聚糖纳米粒较离子凝胶法制备的纳米粒粒径小(P<0.05),且包封率明显提高(P<0.05);中性条件下的累积释放度较pH 5.0小(P<0.05);在pH 5.0醋酸钠缓冲液介质中的粒径和PDI较小,电位较高(P<0.05);对MCF-7细胞的抑制作用随时间增加而逐渐强于游离盐酸阿霉素,且其对肿瘤细胞的抑制效果强于离子凝胶法制备的壳聚糖纳米粒(P<0.05)。结论离子凝胶法和化学交联法联合制备的壳聚糖纳米粒的粒径较小,缓释性较好,对肿瘤细胞的抑制作用较强,为研究盐酸阿霉素新剂型提供了实验依据。

【Objective】To evaluate the potential application of a novel chitosan nanoparticle,which was prepared by the combination of chemical crosslinking and ionic gelation method,by investigating the properties of the nanoparticles.【Methods】Chitosan nanoparticles were prepared by combining the chemical crosslinking and ionic gelation method,with Doxorubicin as a model drug.The entrapment efficiency（EE） and the release rate of novel chitosan nanoparticles were detected by centrifugation and membrane dialysis assay,with chitosan nanoparticles which prepared by ionic gelation method as a control.Also,the size,potential and the polydispersity index（PDI） of novel chitosan nanoparticles in different media and pH were investigated.Then,the viability of the human breast cancer cells MCF-7,which were incubated with novel chitosan nanoparticles were assayed by MTT assay,with nanoparticles and free Doxorubicin as controls.【Results】Novel chitosan nanoparticles were smaller than chitosan nanoparticles（P 0.05）,while the EE was higher（P 0.05）.Novel chitosan nanoparticle at pH 5.0 was smaller than which at pH 7.4,while the release rate was higher（P 0.05）.Among the several mediums,the size and PDI of the novel chitosan nanoparticles in pH 5.0 sodium acetate buffer were the smallest,while the potential was the highest.The inhibition of the novel chitosan nanoparticles to the MCF-7 cells was increasing with time,and further more,it was the highest among the novel chitosan nanoparticles,chitosan nanoparticles and the free doxorubicin（P 0.05）.【Conclusion】 The novel chitosan nanoparticle prepared by the combination of chemical crosslinking and ionic gelation method is small,have a good quality of slow releasing,and strong inhibition to tumor cells.So,it is promising.