Abstract

To determine responses to nanoparticles in a more comprehensive way, current efforts in nanosafety aim at combining the analysis of multiple endpoints and comparing outcomes in different models. To this end, here we used tissue slices from mice as 3D ex vivo models and performed for the first time a comparative study of uptake and impact in liver, lung, and kidney slices exposed under the same conditions to silica, carboxylated and amino-modified polystyrene. In all organs, only exposure to amino-modified polystyrene induced toxicity, with stronger effects in kidneys and lungs. Uptake and distribution studies by confocal microscopy confirmed nanoparticle uptake in all slices, and, in line with what observed in vivo, preferential accumulation in the macrophages. However, uptake levels in kidneys were minimal, despite the strong impact observed when exposed to the amino-modified polystyrene. On the contrary, nanoparticle uptake and accumulation in macrophages were particularly evident in lung slices. Thus, tissue digestion was used to recover all cells from lung slices at different exposure times and to determine by flow cytometry detailed uptake kinetics in lung macrophages and all other cells, confirming higher uptake by the macrophages. Finally, the expression levels of a panel of targets involved in inflammation and macrophage polarization were measured to determine potential effects induced in lung and liver tissue. Overall, this comparative study allowed us to determine uptake and impact of nanoparticles in real tissue and identify important differences in outcomes in the organs in which nanoparticles distribute.

摘要

为了以更全面的方式确定对纳米颗粒的反应，当前在纳米安全方面的努力旨在结合多个端点的分析并比较不同模型中的结果。为此，我们在此使用小鼠的组织切片作为3D离体模型，并首次进行了在相同条件下暴露于二氧化硅，羧化和氨基修饰的肝脏，肺和肾脏切片中摄取和影响的比较研究聚苯乙烯。在所有器官中，只有暴露于氨基改性的聚苯乙烯才会引起毒性，对肾脏和肺部的影响更大。共聚焦显微镜的摄取和分布研究证实了所有切片中的纳米颗粒摄取，并且与体内观察到的一致，在巨噬细胞中优先积累。然而，尽管暴露于氨基改性的聚苯乙烯时观察到强烈的影响，肾脏中的摄取水平却很小。相反，在肺切片中纳米颗粒在巨噬细胞中的摄取和积累尤为明显。因此，组织消化用于在不同的暴露时间从肺切片中回收所有细胞，并通过流式细胞术确定肺巨噬细胞和所有其他细胞的详细吸收动力学，从而证实巨噬细胞的更高吸收。最后，测量涉及炎症和巨噬细胞极化的一组靶标的表达水平，以确定在肺和肝组织中诱导的潜在作用。总体，