Evaluating the uptake and intracellular fate of polystyrene nanoparticles by primary and hepatocyte cell lines in vitro
Evaluating the uptake and intracellular fate of polystyrene nanoparticles by primary and hepatocyte cell lines in vitro
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DOI：
10.1016/j.taap.2009.09.015
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发表时间：
2010-01-01
影响因子：
3.8
通讯作者：
Stone, Vicki
中科院分区：
文献类型：
--
作者：
Johnston, Helinor J.;Semmler-Behnke, Manuela;Stone, Vicki
纳米粒子（NPs）被广泛应用于医药、服装、化妆品和食品等领域。为了促进这种纳米技术的安全发展，必须评估与人类接触有关的潜在不利健康后果。肝脏被认为是NP毒性的靶点，因为在注射、吸入或滴注后，NP会在肝脏内积累。用共聚焦显微镜测定肝细胞对荧光聚苯乙烯羧化颗粒（直径20 nm或200 nm）的摄取；在颗粒暴露期间或暴露后固定细胞内的细胞“活”成像。比较了原代大鼠肝细胞和人肝细胞系（C3A和HepG2）对聚苯乙烯颗粒的摄取情况。肝细胞对颗粒的摄取与大小、时间和血清有关。具体来说，200nm颗粒的摄取是有限的，但20nm NPs从10分钟开始被所有细胞类型内化。10 min时，20 nm NP荧光与微管蛋白骨架染色共定位；30分钟后，NP荧光在细胞内和/或细胞间的结构中划分。考虑了内化NPs的命运，它们不包含在早期的核内体或溶酶体中，而是包含在细胞系的线粒体中。NPs在胆管内的积累是有限的，这表明NPs可以在胆汁中被清除。这与在大鼠体内静脉注射金NPs后在胆汁中被消除的发现是一致的。这些发现大体上在原代大鼠肝细胞和不同的人类肝细胞细胞系之间具有可比性。(C) 2009爱思唯尔公司版权所有。
Nanoparticles (NPs) are being used within diverse applications such as medicines, clothing, cosmetics and food. In order to promote the safe development of such nano technologies it is essential to assess the potential adverse health consequences associated with human exposure. The liver is recognised as a target site for NP toxicity, due to NP accumulation within this organ subsequent to injection, inhalation or instillation. The uptake of fluorescent polystyrene carboxylated particles (20 nm or 200 nm diameter) by hepatocytes was determined using confocal microscopy; with cells imaged "live" during particle exposure or after exposure within fixed cells. Comparisons between the uptake of polystyrene particles by primary rat hepatocytes, and human hepatocyte cell lines (C3A and HepG2) were made. Uptake of particles by hepatocytes was size, time, and serum dependent. Specifically, the uptake of 200 nm particles was limited, but 20 nm NPs were internalised by all cell types from 10 min onwards. At 10 min, 20 nm NP fluorescence co-localised with the tubulin cytoskeleton staining; after 30 min NP fluorescence compartmentalised into structures located within and/or between cells. The fate of internalised NPs was considered and they were not contained within early endosomes or lysosomes, but within mitochondria of cell lines. NPs accumulated within bile canaliculi to a limited extent, which suggests that NPs can be eliminated within bile. This is in keeping with the finding that gold NPs were eliminated in bile following intravenous injection into rats. The findings were, in the main, comparable between primary rat hepatocytes and the different human hepatocyte cell lines. (C) 2009 Elsevier Inc. All rights reserved.