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评估纳米银颗粒对分化 Caco-2 细胞单层的影响,作为肠道屏障模型。

Assessing the effects of silver nanoparticles on monolayers of differentiated Caco-2 cells, as a model of intestinal barrier.

机构信息

Grup de Mutagènesi, Departament de Genètica i de Microbiologia, Facultat de Biociències, Universitat Autònoma de Barcelona, Bellaterra, Spain.

Grup de Mutagènesi, Departament de Genètica i de Microbiologia, Facultat de Biociències, Universitat Autònoma de Barcelona, Bellaterra, Spain; CIBER Epidemiología y Salud Pública, ISCIII, Spain.

出版信息

Food Chem Toxicol. 2018 Jun;116(Pt B):1-10. doi: 10.1016/j.fct.2018.04.008. Epub 2018 Apr 4.

DOI:10.1016/j.fct.2018.04.008
PMID:29626574
Abstract

Since ingestion is one of the main routes of entry of nanoparticles (NPs) in our organism, simple and fast in vitro models of the intestinal barrier can be helpful to evaluate NPs risk. The human colon adenocarcinoma Caco-2 cell line has been extensively used due to its ability to differentiate, forming a well-structured cell monolayer. In this study, we have used these differentiated cells as a model of intestinal barrier to evaluate a wide set of effects caused by exposure to silver nanoparticles (AgNPs) with an average size of 7.74 nm. Different parameters such as toxicity, monolayer integrity and permeability (assessed by changes in cells' morphology and gene expression pattern), internalization (uptake), translocation, and induction of DNA damage (DNA breaks and oxidative DNA damage) were evaluated. No significant effects were observed on the monolayer's integrity/permeability after exposure to silver nanoparticles, although cellular uptake was demonstrated by using confocal microscopy. Despite the observed uptake, no translocation of AgNPs to the basolateral chamber was demonstrated with any of the different experimental approaches used. The genotoxic effects evaluated using the comet assay indicate that, although AgNPs were not able to induce direct DNA breaks, its exposure induced a significant increase in the oxidative DNA damage levels, at non-toxic concentrations.

摘要

由于摄入是纳米颗粒(NPs)进入我们机体的主要途径之一,因此简单快速的体外肠道屏障模型有助于评估 NPs 的风险。人结肠腺癌细胞系 Caco-2 因其能够分化形成结构良好的单层细胞而被广泛应用。在这项研究中,我们使用这些分化细胞作为肠道屏障模型,来评估暴露于平均尺寸为 7.74nm 的银纳米颗粒(AgNPs)所引起的广泛影响。不同的参数,如毒性、单层完整性和通透性(通过细胞形态和基因表达模式的变化来评估)、内化(摄取)、转位和诱导 DNA 损伤(DNA 断裂和氧化 DNA 损伤),都进行了评估。尽管用共聚焦显微镜证实了细胞摄取,但暴露于银纳米颗粒后,单层的完整性/通透性没有观察到显著变化。尽管观察到了摄取,但使用任何不同的实验方法都没有证明 AgNPs 向基底外侧腔的转位。彗星试验评估的遗传毒性效应表明,尽管 AgNPs 不能诱导直接的 DNA 断裂,但在非毒性浓度下,其暴露会导致氧化 DNA 损伤水平显著增加。

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