Impact of nanosilver on various DNA lesions and HPRT gene mutations - effects of charge and surface coating.
Impact of nanosilver on various DNA lesions and HPRT gene mutations - effects of charge and surface coating.
复制标题
纳米层对各种DNA病变和HPRT基因突变的影响 - 电荷和表面涂层的影响。
DOI：
10.1186/s12989-015-0100-x
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发表时间：
2015-07-24
影响因子：
10
通讯作者：
Dusinska M
中科院分区：
文献类型：
--
作者：
Huk A;Izak-Nau E;El Yamani N;Uggerud H;Vadset M;Zasonska B;Duschl A;Dusinska M
本研究的主要目的是根据良好实验室规范（GLP）的标准，在体外研究一套完整表征的银纳米材料（Ag ENMs）与细胞的相互作用，以保证纳米毒理学研究的质量。我们感兴趣的是，采用相同的方法，在相同的尺寸分布、形状和比表面积下，不同的电荷和表面组成是否会产生不同的生物反应。采用一系列方法和毒性终点来研究Ag ENMs与TK6细胞相互作用的影响。以相对生长活性和台盼蓝排除率作为活性测定指标。通过碱性彗星试验检测链断裂和氧化嘌呤来评估遗传毒性。根据OECD方案，采用体外HPRT基因突变试验对V79-4细胞进行了Ag ENMs的致突变性研究。Ag ENM的聚集、溶解以及在细胞内的吸收和分布被研究为Ag ENM毒性的关键方面。除阳性对照和阴性对照外，还包括Ag ENM稳定剂。观察到不同的细胞毒性作用，包括膜损伤、细胞周期阻滞和细胞死亡。Ag ENMs还会引起DNA链断裂、DNA氧化等多种DNA损伤，并引起基因突变。我们发现，与中性或负电荷的Ag ENMs相比，阳性Ag ENMs对细胞毒性和遗传毒性的影响更大，这被认为与它们更容易被细胞吸收以及它们在细胞核和线粒体中的存在有关，这意味着Ag ENMs可能通过直接和间接的机制诱导毒性。我们发现Ag ENMs可能具有细胞毒性、基因毒性和诱变性。我们的HPRT基因突变实验表明，表面化学成分在Ag ENM毒性中起重要作用。
The main goal of this research was to study the interactions of a fully characterized set of silver nanomaterials (Ag ENMs) with cells in vitro, according to the standards of Good Laboratory Practices (GLP), to assure the quality of nanotoxicology research. We were interested in whether Ag ENMs synthesized by the same method, with the same size distribution, shape and specific surface area, but with different charges and surface compositions could give different biological responses. A range of methods and toxicity endpoints were applied to study the impacts of interaction of the Ag ENMs with TK6 cells. As tests of viability, relative growth activity and trypan blue exclusion were applied. Genotoxicity was evaluated by the alkaline comet assay for detection of strand breaks and oxidized purines. The mutagenic potential of Ag ENMs was investigated with the in vitro HPRT gene mutation test on V79-4 cells according to the OECD protocol. Ag ENM agglomeration, dissolution as well as uptake and distribution within the cells were investigated as crucial aspects of Ag ENM toxicity. Ag ENM stabilizers were included in addition to positive and negative controls. Different cytotoxic effects were observed including membrane damage, cell cycle arrest and cell death. Ag ENMs also induced various kinds of DNA damage including strand breaks and DNA oxidation, and caused gene mutation. We found that positive Ag ENMs had greater impact on cyto- and genotoxicity than did Ag ENMs with neutral or negative charge, assumed to be related to their greater uptake into cells and to their presence in the nucleus and mitochondria, implying that Ag ENMs might induce toxicity by both direct and indirect mechanisms. We showed that Ag ENMs could be cytotoxic, genotoxic and mutagenic. Our experiments with the HPRT gene mutation assay demonstrated that surface chemical composition plays a significant role in Ag ENM toxicity.