Lebedová Jana, Hedberg Yolanda S, Odnevall Wallinder Inger, Karlsson Hanna L
Institute of Environmental Medicine, Karolinska Institutet, Nobels väg, Stockholm, Sweden.
RECETOX, Masaryk University, Kamenice, Brno, Czech Republic.
Mutagenesis. 2018 Feb 24;33(1):77-85. doi: 10.1093/mutage/gex027.
Metallic nanoparticles (NPs) are promising nanomaterials used in different technological solutions as well as in consumer products. Silver (Ag), gold (Au) and platinum (Pt) represent three metallic NPs with current or suggested use in different applications. Pt is also used as vehicle exhaust catalyst leading to a possible exposure via inhalation. Despite their use, there is limited data on their genotoxic potential and possible size-dependent effects, particularly for Pt NPs. The aim of this study was to explore size-dependent genotoxicity of these NPs (5 and 50 nm) following exposure of human bronchial epithelial cells. We characterised the NPs and assessed the viability (Alamar blue assay), formation of DNA strand breaks (mini-gel comet assay) and induction of micronucleus (MN) analysed using flow cytometry (in vitro microflow kit). The results confirmed the primary size (5 and 50 nm) but showed agglomeration of all NPs in the serum free medium used. Slight reduced cell viability (tested up to 50 µg/ml) was observed following exposure to the Ag NPs of both particle sizes as well as to the smallest (5 nm) Au NPs. Similarly, at non-cytotoxic concentrations, both 5 and 50 nm-sized Ag NPs, as well as 5 nm-sized Au NPs, increased DNA strand breaks whereas for Pt NPs only the 50 nm size caused a slight increase in DNA damage. No clear induction of MN was observed in any of the doses tested (up to 20 µg/ml). Taken together, by using the comet assay our study shows DNA strand breaks induced by Ag NPs, without any obvious differences in size, whereas effects from Au and Pt NPs were size-dependent in the sense that the 5 nm-sized Au NPs and 50 nm-sized Pt NPs particles were active. No clear induction of MN was observed for the NPs.
金属纳米颗粒(NPs)是很有前景的纳米材料,可用于不同的技术解决方案以及消费品中。银(Ag)、金(Au)和铂(Pt)是三种目前已在不同应用中使用或建议使用的金属纳米颗粒。Pt还用作车辆尾气催化剂,可能导致通过吸入接触。尽管它们有应用,但关于其遗传毒性潜力和可能的尺寸依赖性影响的数据有限,特别是对于Pt纳米颗粒。本研究的目的是探讨这些纳米颗粒(5纳米和50纳米)在暴露于人类支气管上皮细胞后与尺寸相关的遗传毒性。我们对纳米颗粒进行了表征,并评估了细胞活力(alamar蓝分析法)、DNA链断裂的形成(微型凝胶彗星分析法)以及使用流式细胞术(体外微流试剂盒)分析的微核(MN)诱导情况。结果证实了主要尺寸(5纳米和50纳米),但显示在所用的无血清培养基中所有纳米颗粒都发生了团聚。在暴露于两种粒径的Ag纳米颗粒以及最小的(5纳米)Au纳米颗粒后,观察到细胞活力略有降低(测试浓度高达50μg/ml)。同样,在非细胞毒性浓度下,5纳米和50纳米尺寸的Ag纳米颗粒以及5纳米尺寸的Au纳米颗粒都增加了DNA链断裂,而对于Pt纳米颗粒,只有50纳米尺寸导致DNA损伤略有增加。在任何测试剂量(高达20μg/ml)下均未观察到明显的微核诱导。综上所述,通过彗星分析法,我们的研究表明Ag纳米颗粒诱导了DNA链断裂,在尺寸上没有任何明显差异,而Au和Pt纳米颗粒的影响是尺寸依赖性的,即5纳米尺寸的Au纳米颗粒和50纳米尺寸的Pt纳米颗粒具有活性。未观察到纳米颗粒对微核有明显诱导作用。