Vila Laura, García-Rodríguez Alba, Cortés Constanza, Velázquez Antonia, Xamena Noel, Sampayo-Reyes Adriana, Marcos Ricard, Hernández Alba
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.
Chem Biol Interact. 2018 Mar 1;283:38-46. doi: 10.1016/j.cbi.2018.01.018. Epub 2018 Jan 31.
Since ingestion constitute one of the main routes of nanoparticles (NPs) exposure, intestinal cells seems to be a suitable choice to evaluate their potential harmful effects. Caco-2 cells, derived from a human colon adenocarcinoma, have the ability to differentiate forming consistent cell monolayer structures. For these reasons Caco-2 cells, both in their undifferentiated or differentiated state, are extendedly used. We have used well-structured monolayers of differentiated Caco-2 cells, as a model of intestinal barrier, to evaluate potential harmful effects associated to CeONPs exposure via ingestion. Different parameters such as cell toxicity, monolayer integrity and permeability, cell internalization, translocation through the monolayer, and induction of DNA damage were evaluated. No toxic effects of CeONPs were observed, independently of the differentiated state of the Caco-2 cells. In the same way, no effects on the monolayer integrity/permeability were observed. Although important cell uptake was demonstrated in undifferentiated cells (by using confocal microscopy), CeONPs remained mostly attached to the apical membrane in the differentiated cells. In spite of this apparent lack of uptake in differentiated cells, translocation of CeONPs to the basolateral chamber was observed by using confocal microscopy. Finally no genotoxic effects were observed when the comet assay was used, although decreases in the levels of oxidized bases were observed, supporting the antioxidant role of CeONPs.
由于摄入是纳米颗粒（NPs）暴露的主要途径之一，肠道细胞似乎是评估其潜在有害影响的合适选择。源自人结肠腺癌的Caco-2细胞具有分化形成一致细胞单层结构的能力。由于这些原因，Caco-2细胞，无论处于未分化状态还是分化状态，都被广泛使用。我们使用结构良好的分化Caco-2细胞单层作为肠道屏障模型，以评估通过摄入与CeONPs暴露相关的潜在有害影响。评估了不同参数，如细胞毒性、单层完整性和通透性、细胞内化、通过单层的转运以及DNA损伤的诱导。未观察到CeONPs的毒性作用，与Caco-2细胞的分化状态无关。同样，未观察到对单层完整性/通透性的影响。尽管在未分化细胞中证明了重要的细胞摄取（通过共聚焦显微镜观察），但CeONPs在分化细胞中大多仍附着在顶膜上。尽管在分化细胞中明显缺乏摄取，但通过共聚焦显微镜观察到CeONPs转运至基底外侧腔。最后，当使用彗星试验时未观察到遗传毒性作用，尽管观察到氧化碱基水平降低，这支持了CeONPs的抗氧化作用。
