Appropriate In Vitro Methods for Genotoxicity Testing of Silver Nanoparticles.
- Author:
Ha Ryong KIM
1
;
Yong Joo PARK
;
Da Young SHIN
;
Seung Min OH
;
Kyu Hyuck CHUNG
Author Information
1. School of Pharmacy, Sungkyunkwan University, Suwon, Korea. khchung@skku.edu
- Publication Type:In Vitro ; Original Article
- Keywords:
Bacterial reverse mutation test;
Comet assay;
Genotoxicity;
Micronucleus assay;
Silver nanoparticles
- MeSH:
Animals;
Comet Assay;
Cricetinae;
Cricetulus;
Cytochalasin B;
DNA;
Female;
Filtration;
Liver;
Micronucleus Tests;
Mutagenicity Tests*;
Nanoparticles*;
Ovary;
Rats;
Salmonella typhimurium;
Silver*;
Sonication
- From:Environmental Health and Toxicology
2013;28(1):e2013003-
- CountryRepublic of Korea
- Language:English
-
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 microm 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 microg/mL to 10 microg/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.