Gold Nanoparticles Uptake and Cytotoxicity Assessed on Rat Liver Precision-Cut Slices
Gold Nanoparticles Uptake and Cytotoxicity Assessed on Rat Liver Precision-Cut Slices
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DOI：
10.1093/toxsci/kfs150
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发表时间：
2012-07-01
影响因子：
3.8
通讯作者：
Valoti, Massimo
中科院分区：
文献类型：
--
作者：
Dragoni, Stefania;Franco, Giulia;Valoti, Massimo
纳米毒理学领域的一个主要障碍是开发一种能够准确预测体内反应的体外模型。为了解决这一问题，采用大鼠肝脏精密切片来评估包裹了聚乙烯吡咯烷酮(PVP)的5 nm金纳米颗粒(GNPs)在不同浓度下持续24小时对哺乳动物肝脏的影响。在加入GNP后30分钟内，肝细胞内吞饮小泡中可观察到GNP的存在。2 h后，GNPs在切片内的内吞体样小泡内清晰可见，不仅在肝细胞中，而且在位于前两层细胞内的内皮细胞和Kupffer细胞中也可见GNPs。这种摄取没有转化为7-乙氧基香豆素代谢的第一阶段或第二阶段的修饰，也没有改变细胞色素P450对标记底物的活性。此外，尽管GNPs被迅速内化，但没有观察到明显的细胞毒性迹象，通过乳酸脱氢酶释放、甲基噻唑基二苯基四氮唑溴化物的减少和谷胱甘肽水平来评估。总之，大鼠肝切片的使用成功地加强了纳米材料的筛选，并确定了PVP包被的5 nm GNPs与大鼠肝细胞的生物相容性。
A major obstacle in the field of nanotoxicology is the development of an in vitro model that accurately predicts an in vivo response. To address this concern, rat liver precision-cut slices were used to assess the impact of 5-nm gold nanoparticles (GNPs) coated with polyvinylpyrrolidone (PVP) on the mammalian liver, following exposure to different concentrations and for a duration of up to 24 h. The presence of GNPs inside endocytotic vesicles of hepatocytes was appreciable within 30 min of their addition. After 2 h, GNPs were clearly visualized inside endosome-like vesicles within the slice, not only in hepatocytes but also in endothelial and Kupffer cells located within the first two cellular layers. This uptake did not translate into modifications of either phase I or phase II of 7-ethoxycoumarin metabolism or alter activities of cytochrome P450 toward marker substrates. Furthermore, although the GNPs were rapidly internalized, no overt signs of cytotoxicity, assessed through lactate dehydrogenase release, reduction of methylthiazolyldiphenyl tetrazolium bromide, and glutathione levels, were observed. In conclusion, the use of rat liver slices successfully enhanced nanomaterial screening and determined that PVP-coated 5-nm GNPs were biocompatible with rat liver cells.