Despite broad application of magnetic nanoparticles in biomedicine and electronics, only a few in vivo studies on biocompatibility are available. In this study, toxicity of magnetic metal oxide nanoparticles on the respiratory system was examined in vivo by single intratracheal instillation in mice. Bronchoalveolar lavage fluid (BALF) samples were collected for proteome analyses by LC-MS/MS, testing Fe3O4 nanoparticles doped with increasing amounts of cobalt (Fe3O4, CoFe2O4 with an iron to cobalt ratio 5:1, 3:1, 1:3, Co3O4) at two doses (54 μg, 162 μg per animal) and two time points (day 1 and 3 days postinstillation). In discovery phase, in-depth proteome profiling of a few representative samples allowed for comprehensive pathway analyses. Clustering of the 681 differentially expressed proteins (FDR < 0.05) revealed general as well as metal oxide specific responses with an overall strong induction of innate immunity and activation of the complement system. The highest expression increase could be found for a cluster of 39 proteins, which displayed strong dose-dependency to iron oxide and can be attributed to neutrophil extracellular trap (NET) formation. In-depth proteome analysis expanded the knowledge of in vivo NET formation. During screening, all BALF samples of the study (n = 166) were measured label-free as single-injections after a short gradient (21 min) LC separation using the Evosep One system, validating the findings from the discovery and defining protein signatures which enable discrimination of lung inflammation. We demonstrate a proteomics-based toxicity screening with high sample throughput easily transferrable to other nanoparticle types. Data are available via ProteomeXchange with identifier PXD016148.

中文翻译：

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快速和强大的蛋白质组筛查平台可识别肺部对钴铁氧体纳米粒子的中性粒细胞胞外陷阱形成。

尽管磁性纳米颗粒在生物医学和电子学中有广泛的应用，但是仅有少数关于生物相容性的体内研究可用。在这项研究中，通过单次气管内滴注小鼠体内检查了磁性金属氧化物纳米颗粒对呼吸系统的毒性。通过LC-MS / MS收集支气管肺泡灌洗液（BALF）样品进行蛋白质组分析，测试掺有增加钴含量的Fe3O4纳米颗粒（Fe3O4，CoFe2O4铁钴比为5：1、3：1、1：3， Co3O4）两种剂量（54μg，每只动物162μg）和两个时间点（滴注后第1天和第3天）。在发现阶段，对几个代表性样品进行深入的蛋白质组分析可进行全面的途径分析。681种差异表达蛋白的聚类（FDR <0。05）揭示了一般以及金属氧化物的特异性反应，并具有对先天免疫和补体系统活化的总体强诱导作用。对于一组39种蛋白质，可以发现最高的表达增加，这些蛋白质对氧化铁表现出强烈的剂量依赖性，并且可以归因于嗜中性白细胞胞外陷阱（NET）的形成。深入的蛋白质组分析扩展了体内NET形成的知识。在筛选过程中，使用Evosep One系统在短暂的梯度（21分钟）LC分离后，以单次进样的方式对所有研究的BALF样品（n = 166）进行了无标签测量，从而验证了发现的发现并定义了蛋白质标记，可以区分肺部炎症。我们展示了基于蛋白质组学的毒性筛选方法，具有高样品通量，可轻松转移至其他纳米颗粒类型。数据可通过ProteomeXchange获得，其标识符为PXD016148。