甘露糖修饰超顺磁纳米颗粒的制备及体外特性分析

Preparation and in vitro features of mannose-modified superparamagnetic iron oxide nanoparticles

目的探讨以甘露糖(mannose)修饰超顺磁纳米颗粒(superparamagnetic iron oxide nanoparticles,SPION)的制备方法并检测其理学性质,通过体外实验探讨该体系能否被网状内皮系统吞噬。方法采用共沉淀法制备Fe3O4纳米颗粒,然后加入甘露糖溶液,在通氮条件下90℃剧烈搅拌2 h,得到甘露糖包被的SPION。透射电镜测量其大小和分布,傅里叶红外光谱判断甘露糖修饰效果,采用原子吸收分光光度计检测样品中铁含量,体外白细胞吞噬实验检测其能否被白细胞吞噬,细胞毒性实验检测样品的安全性。结果所得样品为球形或类球形,分散性好,核心为Fe3O4晶体,核心粒径为(12.89±3.29)nm,包被甘露糖后的整体颗粒直径不超过20 nm;样品的铁含量为69.84 mg/L;红外吸收光谱测定结果表明甘露糖成功结合到Fe3O4的表面;体外白细胞吞噬实验表明白细胞对甘露糖修饰的SPION的吞噬率较高,细胞毒性实验表明其对细胞的IC50为100.44μg/ml。结论制备的mannose-SPION具有粒径小、分散性好、超顺磁性等优点,容易被吞噬细胞吞噬,且细胞毒性较低。

Objective To explore how to prepare mannose-modified superparamagnetic iron oxide nanopartieles （SPION） and investigate the physical properties in vitro. Methods Co-precipitation was applied to prepare iron oxide nanopartieles. Then mannose was added to the solution to prepare the mannose- SPION under high-speed mechanical stirring at 90 ℃ for 2 h. The size and distribution of the obtained nanoparticles were measured by transmission electron microscopy （TEM）. Fourier transform infrared spectroscopy （FTIS） was used to evaluate the modification of mannose. Atomic absorption spectrophotometry （AAS） was employed to measure the iron content in the samples. Whether the granules were fully engulfed were measured by phagocytic white blood cells in vitro test. The safety of the nanoparticles were measured by cell toxicity test. Results The mannose-SPION was in globular or spherical shape, and in good dispersion. The cores were Fe3O4, with an average core diameter of （ 12.89 ± 3.29 ） nm. The hole diameter of the mannose-SPION was lesser than 20 nm. The iron content of sample was 69.84 mg/L. The result of FTIS indicated that the surface of Fe3O4 had mannose. The phagocytosis test showed that mannose-SPION had higher rate of phagocytosis. The cytotoxicity test displayed that the IC50 of mannose-SPION for cell was 100.44 μg/ml. Conclusion The mannose-SPION possess the characteristics of small size, good dispersion super-paramagnetic response and lower cytotoxicity, and can be phagocytized by phagocyte.