A harmonized and standardized in vitro approach produces reliable results on silver nanoparticles toxicity in different cell lines
A harmonized and standardized in vitro approach produces reliable results on silver nanoparticles toxicity in different cell lines
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DOI：
10.1002/jat.4178
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发表时间：
2021-05-12
影响因子：
3.3
通讯作者：
Barone, Flavia
中科院分区：
文献类型：
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作者：
Andreoli, Cristina;Prota, Valentina;Barone, Flavia
尽管纳米银（AgNP）在不同领域得到了广泛应用，并且进行了大量研究，但迄今为止，对其潜在毒性仍存在许多相互矛盾的结果。在本研究中，使用优化方案和标准化方法对广泛表征的 20 nm AgNP 进行了研究，以测试不同细胞系中的多个毒理学终点。通过动态光散射 (DLS) 测量培养基中 AgNP 的团聚/聚集状态。通过彗星和微核测定评估 BEAS-2B 和 RAW 264.7 细胞的 DNA 和染色体损伤，并通过改良彗星测定和 8-oxodG/8-oxodA 检测评估氧化 DNA 损伤。我们还研究了在有或没有脂多糖 (LPS) 刺激的情况下 RAW 264.7 和 MH-S 细胞中细胞因子释放和 NO 产生的免疫毒性和免疫调节作用。透射电子显微镜 (TEM) 分析用于分析 AgNP 的细胞摄取。我们的结果表明，AgNPs 流体动力学直径的值取决于介质的不同，仅对 BEAS-2B 有一些遗传毒性作用，并且在不存在或存在 LPS 刺激的情况下对 RAW 264.7 和 MH-S 的功能没有或轻微影响。这项研究强调了使用优化方案和多个终点来分析 AgNP 的潜在毒性并获得可靠且可比较的结果的相关性。
Despite the widespread use of silver nanoparticles (AgNPs) in different fields and the amount of investigations available, to date, there are many contradictory results on their potential toxicity. In the present study, extensively characterized 20-nm AgNPs were investigated using optimized protocols and standardized methods to test several toxicological endpoints in different cell lines. The agglomeration/aggregation state of AgNPs in culture media was measured by dynamic light scattering (DLS). DNA and chromosomal damage on BEAS-2B and RAW 264.7 cells were evaluated by comet and micronucleus assays, while oxidative DNA damage by modified comet assay and 8-oxodG/8-oxodA detection. We also investigated immunotoxicity and immunomodulation by cytokine release and NO production in RAW 264.7 and MH-S cells, with or without lipopolysaccharide (LPS) stimulus. Transmission electron microscope (TEM) analysis was used to analyze cellular uptake of AgNPs. Our results indicate different values of AgNPs hydrodynamic diameter depending on the medium, some genotoxic effect just on BEAS-2B and no or slight effects on function of RAW 264.7 and MH-S in absence or presence of LPS stimulus. This study highlights the relevance of using optimized protocols and multiple endpoints to analyze the potential toxicity of AgNPs and to obtain reliable and comparable results.