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      Appropriate In Vitro Methods for Genotoxicity Testing of Silver Nanoparticles

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          Abstract

          Objectives

          We investigated the genotoxic effects of 40-59 nm silver nanoparticles (Ag-NPs) by bacterial reverse mutation assay (Ames test), in vitro comet assay and micronucleus (MN) assay. In particular, we directly compared the effect of cytochalasin B (cytoB) and rat liver homogenate (S9 mix) in the formation of MN by Ag-NPs.

          Methods

          Before testing, we confirmed that Ag-NPs were completely dispersed in the experimental medium by sonication (three times in 1 minute) and filtration (0.2 µm pore size filter), and then we measured their size in a zeta potential analyzer. After that the genotoxicity were measured and especially, S9 mix and with and without cytoB were compared one another in MN assay.

          Results

          Ames test using Salmonella typhimurium TA98, TA100, TA1535 and TA1537 strains revealed that Ag-NPs with or without S9 mix did not display a mutagenic effect. The genotoxicity of Ag-NPs was also evaluated in a mammalian cell system using Chinese hamster ovary cells. The results revealed that Ag-NPs stimulated DNA breakage and MN formation with or without S9 mix in a dose-dependent manner (from 0.01 µg/mL to 10 µg/mL). In particular, MN induction was affected by cytoB.

          Conclusions

          All of our findings, with the exception of the Ames test results, indicate that Ag-NPs show genotoxic effects in mammalian cell system. In addition, present study suggests the potential error due to use of cytoB in genotoxic test of nanoparticles.

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          Author and article information

          Journal
          Environ Health Toxicol
          Environ Health Toxicol
          EHT
          Environmental Health and Toxicology
          The Korean Society of Environmental Health and Toxicology
          2233-6567
          2013
          07 February 2013
          : 28
          : e2013003
          Affiliations
          [1 ]School of Pharmacy, Sungkyunkwan University, Suwon, Korea.
          [2 ]Fusion Technology Laboratory, Hoseo University, Asan, Korea.
          Author notes
          Correspondence: Kyu Hyuck Chung, PhD. 2066 Seobu-ro, Jangan-gu, Suwon 440-746, Korea. Tel: +82-31-290-7714, Fax: +82-31-290-7771, khchung@ 123456skku.edu
          Article
          10.5620/eht.2013.28.e2013003
          3577117
          23440978
          bbb3ae59-f894-4ecd-8520-db5f208fa914
          Copyright © 2013 The Korean Society of Environmental Health and Toxicology

          This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

          History
          : 02 October 2012
          : 06 December 2012
          Categories
          Original Article

          Occupational & Environmental medicine
          bacterial reverse mutation test,comet assay,genotoxicity,micronucleus assay,silver nanoparticles

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