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Impact of nanosilver on various DNA lesions and HPRT gene mutations - effects of charge and surface coating

A. Huk, E. Izak-Nau, N. El Yamani, H. Uggerud, M. Vadset, B. Zasonska, A. Duschl, M. Dusinska,

. 2015 ; 12 (-) : 25. [pub] 20150724

Language English Country England, Great Britain

Document type Comparative Study, Journal Article, Research Support, Non-U.S. Gov't

Persistent link   https://www.medvik.cz/link/bmc16020495

BACKGROUND: The main goal of this research was to study the interactions of a fully characterized set of silver nanomaterials (Ag ENMs) with cells in vitro, according to the standards of Good Laboratory Practices (GLP), to assure the quality of nanotoxicology research. We were interested in whether Ag ENMs synthesized by the same method, with the same size distribution, shape and specific surface area, but with different charges and surface compositions could give different biological responses. METHODS: A range of methods and toxicity endpoints were applied to study the impacts of interaction of the Ag ENMs with TK6 cells. As tests of viability, relative growth activity and trypan blue exclusion were applied. Genotoxicity was evaluated by the alkaline comet assay for detection of strand breaks and oxidized purines. The mutagenic potential of Ag ENMs was investigated with the in vitro HPRT gene mutation test on V79-4 cells according to the OECD protocol. Ag ENM agglomeration, dissolution as well as uptake and distribution within the cells were investigated as crucial aspects of Ag ENM toxicity. Ag ENM stabilizers were included in addition to positive and negative controls. RESULTS: Different cytotoxic effects were observed including membrane damage, cell cycle arrest and cell death. Ag ENMs also induced various kinds of DNA damage including strand breaks and DNA oxidation, and caused gene mutation. We found that positive Ag ENMs had greater impact on cyto- and genotoxicity than did Ag ENMs with neutral or negative charge, assumed to be related to their greater uptake into cells and to their presence in the nucleus and mitochondria, implying that Ag ENMs might induce toxicity by both direct and indirect mechanisms. CONCLUSION: We showed that Ag ENMs could be cytotoxic, genotoxic and mutagenic. Our experiments with the HPRT gene mutation assay demonstrated that surface chemical composition plays a significant role in Ag ENM toxicity.

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$a Huk, Anna $u Health Effects Laboratory, MILK, NILU, Kjeller, Norway. annahuk8@gmail.com. Department of Molecular Biology, University of Salzburg, Salzburg, Austria. annahuk8@gmail.com.
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$a BACKGROUND: The main goal of this research was to study the interactions of a fully characterized set of silver nanomaterials (Ag ENMs) with cells in vitro, according to the standards of Good Laboratory Practices (GLP), to assure the quality of nanotoxicology research. We were interested in whether Ag ENMs synthesized by the same method, with the same size distribution, shape and specific surface area, but with different charges and surface compositions could give different biological responses. METHODS: A range of methods and toxicity endpoints were applied to study the impacts of interaction of the Ag ENMs with TK6 cells. As tests of viability, relative growth activity and trypan blue exclusion were applied. Genotoxicity was evaluated by the alkaline comet assay for detection of strand breaks and oxidized purines. The mutagenic potential of Ag ENMs was investigated with the in vitro HPRT gene mutation test on V79-4 cells according to the OECD protocol. Ag ENM agglomeration, dissolution as well as uptake and distribution within the cells were investigated as crucial aspects of Ag ENM toxicity. Ag ENM stabilizers were included in addition to positive and negative controls. RESULTS: Different cytotoxic effects were observed including membrane damage, cell cycle arrest and cell death. Ag ENMs also induced various kinds of DNA damage including strand breaks and DNA oxidation, and caused gene mutation. We found that positive Ag ENMs had greater impact on cyto- and genotoxicity than did Ag ENMs with neutral or negative charge, assumed to be related to their greater uptake into cells and to their presence in the nucleus and mitochondria, implying that Ag ENMs might induce toxicity by both direct and indirect mechanisms. CONCLUSION: We showed that Ag ENMs could be cytotoxic, genotoxic and mutagenic. Our experiments with the HPRT gene mutation assay demonstrated that surface chemical composition plays a significant role in Ag ENM toxicity.
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$a Izak-Nau, Emilia $u Department of Molecular Biology, University of Salzburg, Salzburg, Austria. izak.e@hotmail.com. Bayer Technology Services GmbH, Leverkusen, Germany. izak.e@hotmail.com.
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$a El Yamani, Naouale $u Health Effects Laboratory, MILK, NILU, Kjeller, Norway. elyamani.naouale@nilu.no.
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$a Uggerud, Hilde $u Inorganic Group, MILK, NILU, Kjeller, Norway. hilde.thelle.uggerud@nilu.no.
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$a Vadset, Marit $u Inorganic Group, MILK, NILU, Kjeller, Norway. marit.vadset@nilu.no.
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$a Zasonska, Beata $u Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, Prague, Czech Republic. beata.zasonska@gmail.com.
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$a Duschl, Albert $u Department of Molecular Biology, University of Salzburg, Salzburg, Austria. albert.duschl@sbg.ac.at.
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$a Dusinska, Maria $u Health Effects Laboratory, MILK, NILU, Kjeller, Norway. mdu@nilu.no.
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