Department of Physics and LASSP, Cornell University, Ithaca, NY, 14853, USA.
Center for Brain Science, RIKEN, Saitama, 351-0198, Japan.
Sci Rep. 2020 Jul 8;10(1):11239. doi: 10.1038/s41598-020-67724-w.
Recent technological advances have introduced diverse engineered nanoparticles (ENPs) into our air, water, medicine, cosmetics, clothing, and food. However, the health and environmental effects of these increasingly common ENPs are still not well understood. In particular, potential neurological effects are one of the most poorly understood areas of nanoparticle toxicology (nanotoxicology), in that low-to-moderate neurotoxicity can be subtle and difficult to measure. Culturing primary neuron explants on planar microelectrode arrays (MEAs) has emerged as one of the most promising in vitro techniques with which to study neuro-nanotoxicology, as MEAs enable the fluorescent tracking of nanoparticles together with neuronal electrical activity recording at the submillisecond time scale, enabling the resolution of individual action potentials. Here we examine the dose-dependent neurotoxicity of dextran-coated iron oxide nanoparticles (dIONPs), a common type of functionalized ENP used in biomedical applications, on cultured primary neurons harvested from postnatal day 0-1 mouse brains. A range of dIONP concentrations (5-40 µg/ml) were added to neuron cultures, and cells were plated either onto well plates for live cell, fluorescent reactive oxidative species (ROS) and viability observations, or onto planar microelectrode arrays (MEAs) for electrophysiological measurements. Below 10 µg/ml, there were no dose-dependent cellular ROS increases or effects in MEA bursting behavior at sub-lethal dosages. However, above 20 µg/ml, cell death was obvious and widespread. Our findings demonstrate a significant dIONP toxicity in cultured neurons at concentrations previously reported to be safe for stem cells and other non-neuronal cell types.
近年来,各种工程纳米粒子(ENPs)被引入我们的空气、水、药物、化妆品、衣物和食品中。然而,这些越来越常见的 ENPs 的健康和环境影响仍未得到很好的理解。特别是,潜在的神经效应是纳米颗粒毒理学(纳米毒理学)中最不为人知的领域之一,因为低至中等程度的神经毒性可能很微妙,难以测量。在平面微电极阵列(MEA)上培养原代神经元外植体已成为研究神经纳米毒理学最有前途的体外技术之一,因为 MEA 可以实现纳米颗粒的荧光跟踪以及亚毫秒时间尺度的神经元电活性记录,从而可以分辨单个动作电位。在这里,我们研究了葡聚糖包覆的氧化铁纳米粒子(dIONP)的剂量依赖性神经毒性,dIONP 是一种用于生物医学应用的常见功能化 ENP。我们从出生后 0-1 天的小鼠脑中收获培养的原代神经元,并将一系列 dIONP 浓度(5-40μg/ml)添加到神经元培养物中。细胞要么接种到微孔板上进行活细胞、荧光活性氧(ROS)和活力观察,要么接种到平面微电极阵列(MEA)上进行电生理测量。低于 10μg/ml 时,在亚致死剂量下,没有剂量依赖性的细胞 ROS 增加或 MEA 爆发行为的影响。然而,高于 20μg/ml 时,细胞死亡明显且广泛。我们的研究结果表明,在先前报道对干细胞和其他非神经元细胞类型安全的浓度下,培养神经元中的 dIONP 毒性显著。