聚乙烯吡咯烷酮包覆的银纳米粒子对 BEAS-2B 细胞的遗传毒性。
Genotoxicity of polyvinylpyrrolidone-coated silver nanoparticles in BEAS 2B cells.
机构信息
Nanosafety Research Center and Safe New Technologies, Work Environment Development, Finnish Institute of Occupational Health, Helsinki, Finland.
出版信息
Toxicology. 2013 Nov 8;313(1):38-48. doi: 10.1016/j.tox.2012.09.014. Epub 2012 Nov 8.
Silver nanoparticles (AgNPs) are widely utilized in various consumer products and medical devices, especially due to their antimicrobial properties. However, several studies have associated these particles with toxic effects, such as inflammation and oxidative stress in vivo and cytotoxic and genotoxic effects in vitro. Here, we assessed the genotoxic effects of AgNPs coated with polyvinylpyrrolidone (PVP) (average diameter 42.5±14.5 nm) on human bronchial epithelial BEAS 2B cells in vitro. AgNPs were dispersed in bronchial epithelial growth medium (BEGM) with 0.6 mg/ml bovine serum albumin (BSA). The AgNP were partially well-dispersed in the medium and only limited amounts (ca. 0.02 μg Ag(+) ion/l) could be dissolved after 24h. The zeta-potential of the AgNPs was found to be highly negative in pure water but was at least partially neutralized in BEGM with 0.6 mg BSA/ml. Cytotoxicity was measured by cell number count utilizing Trypan Blue exclusion and by an ATP-based luminescence cell viability assay. Genotoxicity was assessed by the alkaline single cell gel electrophoresis (comet) assay, the cytokinesis-block micronucleus (MN) assay, and the chromosomal aberration (CA) assay. The cells were exposed to various doses (0.5-48 μg/cm(2) corresponding to 2.5-240 μg/ml) of AgNPs for 4 and 24 h in the comet assay, for 48 h in the MN assay, and for 24 and 48 h in the CA assay. DNA damage measured by the percent of DNA in comet tail was induced in a dose-dependent manner after both the 4-h and the 24-h exposures to AgNPs, with a statistically significant increase starting at 16 μg/cm(2) (corresponding to 60.8 μg/ml) and doubling of the percentage of DNA in tail at 48 μg/cm(2). However, no induction of MN or CAs was observed at any of the doses or time points. The lack of induction of chromosome damage by the PVP-coated AgNPs is possibly due to the coating which may protect the cells from direct interaction with the AgNPs, either by reducing ion leaching from the particles or by causing extensive agglomeration of the nanoparticles, with a possible reduction of the cellular uptake.
纳米银颗粒(AgNPs)广泛应用于各种消费品和医疗器械中,这主要是由于其具有抗菌特性。然而,有几项研究表明,这些颗粒会在体内引起炎症和氧化应激等毒性作用,以及在体外产生细胞毒性和遗传毒性。在这里,我们评估了涂覆有聚乙烯吡咯烷酮(PVP)的纳米银颗粒(平均直径 42.5±14.5nm)对体外人支气管上皮 BEAS-2B 细胞的遗传毒性。AgNPs 分散在支气管上皮生长培养基(BEGM)中,其中含有 0.6mg/ml 牛血清白蛋白(BSA)。AgNP 在培养基中部分分散良好,仅有限量(约 0.02μgAg(+)离子/l)在 24 小时后可以溶解。AgNP 的动电位在纯水中呈高度负电性,但在含有 0.6mg/mlBSA 的 BEGM 中至少部分中和。通过台盼蓝排斥法和基于 ATP 的荧光细胞活力测定法测量细胞毒性。通过碱性单细胞凝胶电泳(彗星)试验、胞质分裂阻断微核(MN)试验和染色体畸变(CA)试验评估遗传毒性。细胞分别暴露于各种剂量(0.5-48μg/cm(2)相当于 2.5-240μg/ml)的 AgNPs 中 4 小时和 24 小时进行彗星试验,48 小时进行 MN 试验,24 小时和 48 小时进行 CA 试验。通过彗星试验中 DNA 在彗星尾部的百分比测量,在 4 小时和 24 小时暴露于 AgNPs 后,均呈现出剂量依赖性的 DNA 损伤,从 16μg/cm(2)(相当于 60.8μg/ml)开始呈统计学显著增加,并且在 48μg/cm(2)时,尾部的 DNA 百分比增加了一倍。然而,在任何剂量或时间点均未观察到 MN 或 CA 的诱导。涂覆有 PVP 的 AgNPs 不会引起染色体损伤,这可能是由于涂层的作用,它可能通过减少颗粒中的离子浸出或通过引起纳米颗粒的广泛团聚,从而减少细胞摄取,从而使细胞免受与 AgNPs 的直接相互作用。