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目的体外评价自制羧甲基壳聚糖超顺磁氧化铁纳米粒(OCMCS-SPIO-NPs)的细胞毒性和逃避巨噬细胞的吞噬能力。方法以菲立磁和未包被的超顺磁氧化铁纳米粒为对照,采用四唑盐(MTT)比色法考察OCMCS-SPIO-NPs对LO2细胞(正常肝细胞株)和A549(人肺腺癌细胞株)的细胞毒性;用菲洛嗪法及普鲁士蓝法考察OCMCS-SPIO-NPs纳米粒并评价其体外的抗吞噬能力。结果 SPIO-NPs经羧甲基壳聚糖共价修饰后,对LO2和A549细胞的细胞毒性明显降低,OCMCS-SPIO-NPs的细胞毒性和dextran-SPIO的细胞毒性无显著性差异(P>0.05),其细胞毒性与培养基中SPIO的浓度呈正相关;与3种SPIO纳米粒孵化24h后,RAW264.7细胞内铁含量随培养基中SPIO的含量增加而增加,细胞内铁含量依次为:未包被SPIO-NPs>dextran-SPIO-NPs>CMCS-SPIO-NPs组(P<0.05)。结论超顺氧化铁纳米粒经羧甲基壳聚糖共价修饰后能显著降低细胞毒性和吞噬细胞摄取,提高了生物相容性,显著降低了巨噬细胞对其的摄取。
OBJECTIVE: To evaluate the cytotoxicity and escape phagocytosis of macrophages in vitro by self-made carboxymethyl chitosan superparamagnetic iron oxide nanoparticles (OCMCS-SPIO-NPs). Methods MTT assay was used to examine the effect of OCMCS-SPIO-NPs on LO2 cells (normal liver cell line) and A549 (human lung Adenocarcinoma cell line). The morphological changes of OCMCS-SPIO-NPs nanoparticles were evaluated by the phenothiazine method and Prussian blue method and their anti-phagocytic activity in vitro was evaluated. Results After covalently modified with carboxymethyl chitosan, the cytotoxicity of SPIO-NPs on LO2 and A549 cells was significantly reduced. The cytotoxicity of OCMCS-SPIO-NPs and the cytotoxicity of dextran-SPIO were not significantly different (P> 0.05 ). The cytotoxicity was positively correlated with the concentration of SPIO in the culture medium. After incubation with the three kinds of SPIO nanoparticles for 24 h, the content of iron in RAW264.7 cells increased with the increase of the SPIO content in the culture medium. The intracellular iron contents were as follows: SPIO-NPs> dextran-SPIO-NPs> CMCS-SPIO-NPs group were not coated (P <0.05). CONCLUSION: Covalent modification of the super-iron oxide nanoparticles by carboxymethyl chitosan can significantly reduce cytotoxicity and phagocyte uptake, improve biocompatibility and significantly uptake macrophages.