Platel Anne, Carpentier Rodolphe, Becart Elodie, Mordacq Gwendoline, Betbeder Didier, Nesslany Fabrice
Université de Lille 2, 59000, Lille, France.
Institut Pasteur de Lille, Laboratoire de Toxicologie Génétique, 1 rue du Professeur Calmette, BP 245, 59019, Lille, France.
J Appl Toxicol. 2016 Mar;36(3):434-44. doi: 10.1002/jat.3247. Epub 2015 Oct 21.
With the ongoing commercialization of nanotechnology products, human exposure to nanoparticles (NPs) is set to increase dramatically and an evaluation of their potential adverse effects is essential. Surface charge, among other physico-chemicals parameters, is a key criterion that should be considered when using a definition for nanomaterials in a regulatory context. It has recently been recognized as an important factor in determining the toxicity of NPs; however, a complete understanding of the mechanisms involved is still lacking. In this context, the aim of the present study was to investigate the influence of the surface charge modification of NPs on in vitro toxicity assays. Poly(lactic-co-glycolic acid) (PLGA) nanoparticles bearing different surface charges, positive(+), neutral(n) or negative(-), were synthesized. In vitro genotoxicity assays (micronucleus and comet assays) coupled with an assessment of cytotoxicity, were performed in different cell lines (L5178Y mouse lymphoma cells, TK6 human B-lymphoblastoid cells and 16HBE14o- human bronchial epithelial cells). Reactive oxygen species (ROS) production and endocytosis studies were also performed. Our results showed that PLGA(+) NPs were cytotoxic. They are endocytosed by the clathrin pathway and induced ROS in the three cell lines. They led to chromosomal aberrations without primary DNA damage in 16HBE14o- cells, suggesting that aneuploidy may be considered as an important biomarker when assessing the genotoxic potential of NPs. Moreover, 16HBE14o- cells seem to be more suitable for the in vitro screening of inhaled NPs than the regulatory L5178Y and TK6 cells.
随着纳米技术产品的不断商业化，人类对纳米颗粒（NPs）的接触预计将急剧增加，因此评估其潜在的不利影响至关重要。在监管背景下使用纳米材料定义时，表面电荷以及其他物理化学参数是应考虑的关键标准。最近，表面电荷已被认为是决定纳米颗粒毒性的重要因素；然而，对其中涉及的机制仍缺乏全面了解。在此背景下，本研究的目的是探讨纳米颗粒表面电荷修饰对体外毒性测定的影响。合成了带有不同表面电荷，正（+）、中性（n）或负（-）的聚乳酸-乙醇酸共聚物（PLGA）纳米颗粒。在不同细胞系（L5178Y小鼠淋巴瘤细胞、TK6人B淋巴母细胞和16HBE14o-人支气管上皮细胞）中进行了体外遗传毒性测定（微核试验和彗星试验）以及细胞毒性评估。还进行了活性氧（ROS）产生和内吞作用研究。我们的结果表明，PLGA（+）纳米颗粒具有细胞毒性。它们通过网格蛋白途径被内吞，并在三种细胞系中诱导ROS产生。它们在16HBE14o-细胞中导致染色体畸变而无原发性DNA损伤，这表明在评估纳米颗粒的遗传毒性潜力时，非整倍体可能被视为一个重要的生物标志物。此外，与监管用的L5178Y和TK6细胞相比，16HBE14o-细胞似乎更适合用于吸入性纳米颗粒的体外筛选。
