Ávalos A, Haza A I, Morales P
Departamento de Nutrición, Bromatología y Tecnología de los Alimentos. Facultad de Veterinaria, Universidad Complutense de Madrid, Spain.
Folia Biol (Praha). 2015;61(1):33-42. doi: 10.14712/fb2015061010033.
Many classes of silver nanoparticles (AgNPs) have been synthesized and widely applied, but no conclusive information on their potential cytotoxicity and genotoxicity mechanisms is available. Therefore, the purpose of this study was to compare the potential genotoxic effects (DNA strand breaks and oxidative DNA damage) of 4.7 nm coated and 42 nm uncoated AgNPs, using the comet assay, in four relevant human cell lines (hepatoma, leukaemia, and dermal and pulmonary fibroblasts) in order to understand the impact of such nanomaterials on cellular DNA. The results indicated that in all cell lines tested, 4.7 nm coated (0.1-1.6 μg ml⁻¹) and 42 nm uncoated (0.1-6.7 μg ml⁻¹) AgNPs increased DNA strand breaks in a dose- and size-dependent manner following 24 h treatment, the smaller AgNPs being more genotoxic. Human pulmonary fibroblasts showed the highest sensitivity to the AgNPs. A modified comet assay using endonuclease III and formamidopyrimidine- DNA glycosylase restriction enzymes showed that in tumoral and normal human dermal fibroblasts, pyrimidines and purines were oxidatively damaged by both AgNPs, but the damage was not size-dependent. However, in human pulmonary fibroblasts, no oxidative damage was observed after treatment with 42 nm AgNPs. In conclusion, both AgNP sizes induced DNA damage in human cells, and this damage could be related to oxidative stress.
人们已经合成并广泛应用了多种类型的银纳米颗粒(AgNP),但关于其潜在细胞毒性和基因毒性机制尚无确凿信息。因此,本研究旨在使用彗星试验,比较4.7纳米包被的和42纳米未包被的AgNP在四种相关人类细胞系(肝癌细胞系、白血病细胞系以及皮肤和肺成纤维细胞系)中的潜在基因毒性效应(DNA链断裂和氧化性DNA损伤),以了解此类纳米材料对细胞DNA的影响。结果表明,在所有测试细胞系中,经24小时处理后,4.7纳米包被的(0.1 - 1.6微克/毫升)和42纳米未包被的(0.1 - 6.7微克/毫升)AgNP均以剂量和尺寸依赖性方式增加了DNA链断裂,较小的AgNP具有更强的基因毒性。人肺成纤维细胞对AgNP表现出最高的敏感性。使用核酸内切酶III和甲酰胺嘧啶 - DNA糖基化酶限制酶的改良彗星试验表明,在肿瘤性和正常人类皮肤成纤维细胞中,两种AgNP均使嘧啶和嘌呤发生氧化性损伤,但损伤并不依赖于尺寸。然而,在用42纳米AgNP处理后人肺成纤维细胞中未观察到氧化性损伤。总之,两种尺寸的AgNP均在人类细胞中诱导了DNA损伤,且这种损伤可能与氧化应激有关。