Uptake and toxicity of copper oxide nanoparticles in cultured primary brain astrocytes
Uptake and toxicity of copper oxide nanoparticles in cultured primary brain astrocytes
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DOI：
10.3109/17435390.2013.829591
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发表时间：
2014-11-01
期刊：
影响因子：
5
通讯作者：
Dringen, Ralf
中科院分区：
文献类型：
--
作者：
Bulcke, Felix;Thiel, Karsten;Dringen, Ralf
为了测试脑细胞暴露于氧化铜纳米颗粒（CuO-NP）的后果，我们合成并表征了二巯基琥珀酸酯涂覆的CuO-NP。通过透射电子显微镜测定，这些颗粒具有约5 nm的直径，而它们在水性分散体中的平均流体动力学直径为136 +/-4nm。在含有10%胎牛血清的细胞培养基中的分散体将流体动力学直径增加至178 +/-12 nm，并将颗粒的ζ电位从-49 +/-7 mV（在水中）移动至-10 +/-3 mV。暴露于培养的原代脑星形胶质细胞的CuO纳米颗粒增加细胞铜水平，并损害细胞活力的时间，浓度和温度依赖性的方式。以高于100 μ M铜（6.4 μ g/ml）的浓度施用CuO-NP严重损害细胞的生存力，如通过降低的3-（4，5-二甲基噻唑-2-基）-2，5-二苯基溴化四唑还原能力、降低的细胞乳酸脱氢酶活性和增加的荧光染料碘化丙啶的膜渗透性所证明的。暴露于CuO-NP的星形胶质细胞的铜内化以及细胞毒性与已经用铜盐孵育的细胞所观察到的相似。CuO-NP诱导的毒性伴随着细胞中活性氧（ROS）产生的增加。在细胞可渗透的铜螯合剂四硫代钼酸盐的存在下，CuO-NP处理的星形胶质细胞中的ROS形成和细胞毒性均降低。这些数据表明CuO-NP被培养的星形胶质细胞摄取，并表明过量的内化CuO-NP通过加速ROS的形成而引起细胞毒性。
To test for consequences of an exposure of brain cells to copper oxide nanoparticles (CuO-NPs), we synthesised and characterised dimercaptosuccinate-coated CuO-NPs. These particles had a diameter of around 5 nm as determined by transmission electron microscopy, while their average hydrodynamic diameter in aqueous dispersion was 136 +/- 4 nm. Dispersion in cell-culture medium containing 10% fetal calf serum increased the hydrodynamic diameter to 178 +/- 12 nm and shifted the zeta potential of the particles from -49 +/- 7 mV (in water) to -10 +/- 3 mV. Exposure of cultured primary brain astrocytes to CuO-NPs increased the cellular copper levels and compromised the cell viability in a time-, concentration- and temperature-dependent manner. Application of CuO-NPs in concentrations above 100 mu M copper (6.4 mu g/ml) severely compromised the viability of the cells, as demonstrated by a lowered 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide reduction capacity, a lowered cellular lactate dehydrogenase activity and an increased membrane permeability for the fluorescent dye propidium iodide. Copper internalisation as well as cell toxicity of astrocytes exposed to CuO-NPs were similar to that observed for cells that had been incubated with copper salts. The CuO-NP-induced toxicity was accompanied by an increase in the generation of reactive oxygen species (ROS) in the cells. Both, ROS formation and cell toxicity in CuO-NP-treated astrocytes, were lowered in the presence of the cell-permeable copper chelator tetrathiomolybdate. These data demonstrate that CuO-NPs are taken up by cultured astrocytes and suggest that excess of internalised CuO-NPs cause cell toxicity by accelerating the formation of ROS.