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目的:制备柚皮素壳聚糖纳米粒,初步探讨其对人肺腺癌细胞A549的细胞毒性和细胞摄取。方法:以壳聚糖和鱼精蛋白作为载体材料,采用离子胶凝法制备柚皮素壳聚糖纳米粒,透射电镜(TEM)观察其形态,马尔文激光粒度仪测定其粒径、分散度(PDI)和Zeta电位,离心法测定其包封率和载药量,采用恒温振荡水浴法对柚皮素壳聚糖纳米粒进行体外释放度研究,最后采用人肺癌细胞系A549细胞进行了细胞毒性、细胞摄取研究。结果:柚皮素壳聚糖纳米粒为球形或类球形粒子,结构完整,大小均一、球形度好,分散均匀,PDI、粒径、Zeta电位和包封率分别为0.268,139 nm、+15.7 mV和83.34%,柚皮素壳聚糖纳米粒体外释放呈缓释,24 h累积释放量达到了80%以上,体外释药过程用Higuchi方程拟合较好。MTT试验显示不同浓度的壳聚糖纳米粒和细胞作用72 h后,细胞活力均大于95%,本文所制备的壳聚糖纳米粒无细胞毒性。细胞摄取试验表明载FITC的壳聚糖纳米粒和A549细胞作用3 h后,可明显看到大量带绿色荧光的纳米粒穿过细胞膜进入细胞。结论:离子凝胶法成功制得粒径较小的柚皮素壳聚糖纳米粒,具有缓释性好,毒性小,壳聚糖纳米粒摄取率较高,可大大提高药物的利用率,具有广泛的应用前景。
OBJECTIVE: To prepare naringenin chitosan nanoparticles for preliminary study of cytotoxicity and cell uptake into human lung adenocarcinoma A549 cells. METHODS: Chitosan and protamine were used as carrier materials, and naringenin chitosan nanoparticles were prepared by ion-gel method. The morphology was observed by transmission electron microscopy (TEM). The particle size and dispersity (PDI) and Zeta potential. The entrapment efficiency and drug loading were measured by centrifugation. The in vitro release of naringenin chitosan nanoparticles was studied by constant temperature shaking water bath. Finally, human lung cancer cell line A549 Toxicity, cellular uptake studies. Results: Naringenin-loaded chitosan nanoparticles were spherical or spherical particles with a complete structure, uniform size, good sphericity and uniform dispersion. The PDI, particle size, Zeta potential and entrapment efficiency were 0.268 and 139 nm, respectively mV and 83.34% respectively. The release of naringenin chitosan nanoparticles in vitro was sustained release, and the accumulated release amount reached more than 80% in 24 h. Higuchi equation was fit in vitro. MTT assay showed that the cell viability of all the chitosan nanoparticles with different concentrations was higher than 95% after 72 h. The chitosan nanoparticles prepared in this study showed no cytotoxicity. Cell uptake assays showed that after a 3-hour incubation of FITC-loaded chitosan nanoparticles with A549 cells, it was evident that a large number of green fluorescent nanoparticles passed through the cell membrane into the cells. CONCLUSION: The naringenin chitosan nanoparticles with smaller particle size can be successfully prepared by ion-gel method with good sustained-release properties, low toxicity and high uptake rates of chitosan nanoparticles, which can greatly improve the utilization rate of drugs, Has a wide range of applications.