不同浓度超顺磁性氧化铁纳米粒子对小鼠RAW264.7巨噬细胞的影响

Influence of superparamagnetic iron oxide nano-particles in different concentration on macrophage line RAW264.7 in mice

目的评价不同浓度超顺磁性氧化铁纳米粒子(superparamagnetic iron oxides,SPIO)对小鼠RAW264.7巨噬细胞的细胞活性及吞噬功能的影响。方法常规方法培养小鼠RAW264.7巨噬细胞,应用不同浓度SPIO(0μg/ml、14μg/ml、28μg/ml、56μg/ml、84μg/ml、140μg/ml、280μg/ml、560μg/ml、840μg/ml)标记小鼠RAW264.7巨噬细胞,采用普鲁士蓝染色检测细胞标记率,台盼蓝染色检测细胞活性,四唑盐(MTT)比色实验检测细胞增殖能力,中性红吞噬实验检测细胞吞噬功能。结果 SPIO含铁浓度84μg/ml标记24小时,细胞标记率可以达到100%;以后随SPIO浓度的增加,细胞内吞噬的铁颗粒增加,当SPIO含铁浓度为280μg/ml时,细胞内吞噬铁颗粒达到饱和;当SPIO含铁浓度大于280μg/ml时,细胞活性下降,吞噬能力降低;当SPIO含铁浓度大于140μg/ml时,细胞增殖能力下降。结论 SPIO含铁浓度为(84～140)μg/ml标记24h,细胞的标记率为100%并且不影响细胞活性、细胞增殖能力及吞噬能力。

Objective To review the influence of superparamagnetic iron oxide nano-particles （ SPIO ） in different concentration on the cell viability and phagocytic capacity of macrophage line RAW264.7 in mice. Methods RAW264.7 were cultured by using routine method, and then labeled with SPIO in different concentration （ 0μ g/mL, 14 μ g/mL, 28μ g/mL, 56 μ g/mL, 84 μ g/mL, 140 μ g/mL, 280 μ g/mL, 560 μ g/mL and 840 μ g/mL ） . The labeling yield was detected by using Prussian blue staining, cell viability was detected by using trypan blue, cell reproductive capacity was detected by using MTT assay, and cell phagocytic capacity was detected by using neutral red phagocytosis test. Results The labeling rate reached to 100% after labeling for 24 hours in SPIO with iron concentration of 84μ g/mL, and then as iron concentration in SPIO increased, the iron particles phagocytosed by RAW264.7 increased. When the iron concentration in SPIO was 280 μ g/mL, the iron particles phagocytosed by RAW264.7 reached to saturation. When the iron concentration in SPIO was over 280 μ g/mL, cell viability and phagocytic capacity decreased, and the iron concentration in SPIO was over 140 μ g/mL, cell reproductive capacity decreased. Conclusion When the iron concentration in SPIO is （ 84-140 ） μ g/mL and after labeling RAW264.7 for 24 hours, the labeling rate is 100% and cell viability, cell reproductive capacity and phagocytic capacity are not influenced.