Assessing Genotoxicity of Ten Different Engineered Nanomaterials by the Novel Semi-Automated FADU Assay and the Alkaline Comet Assay
Increased engineered nanomaterial (ENM) production and incorporation in consumer and biomedical products has raised concerns about the potential adverse effects. The DNA damaging capacity is of particular importance since damaged genetic material can lead to carcinogenesis. Consequently, reliable an...
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| Format: | Article |
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MDPI AG
2022-01-01
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| Series: | Nanomaterials |
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| Online Access: | https://www.mdpi.com/2079-4991/12/2/220 |
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| author | Sarah May Cordula Hirsch Alexandra Rippl Alexander Bürkle Peter Wick |
| author_facet | Sarah May Cordula Hirsch Alexandra Rippl Alexander Bürkle Peter Wick |
| author_sort | Sarah May |
| collection | DOAJ |
| description | Increased engineered nanomaterial (ENM) production and incorporation in consumer and biomedical products has raised concerns about the potential adverse effects. The DNA damaging capacity is of particular importance since damaged genetic material can lead to carcinogenesis. Consequently, reliable and robust in vitro studies assessing ENM genotoxicity are of great value. We utilized two complementary assays based on different measurement principles: (1) comet assay and (2) FADU (fluorimetric detection of alkaline DNA unwinding) assay. Assessing cell viability ruled out false-positive results due to DNA fragmentation during cell death. Potential structure–activity relationships of 10 ENMs were investigated: three silica nanoparticles (SiO<sub>2</sub>-NP) with varying degrees of porosity, titanium dioxide (TiO<sub>2</sub>-NP), polystyrene (PS-NP), zinc oxide (ZnO-NP), gold (Au-NP), graphene oxide (GO) and two multi-walled carbon nanotubes (MWNT). SiO<sub>2</sub>-NPs, TiO<sub>2</sub>-NP and GO were neither cytotoxic nor genotoxic to Jurkat E6-I cells. Quantitative interference corrections derived from GO results can make the FADU assay a promising screening tool for a variety of ENMs. MWNT merely induced cytotoxicity, while dose- and time-dependent cytotoxicity of PS-NP was accompanied by DNA fragmentation. Hence, PS-NP served to benchmark threshold levels of cytotoxicity at which DNA fragmentation was expected. Considering all controls revealed the true genotoxicity for Au-NP and ZnO-NP at early time points. |
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| institution | Directory Open Access Journal |
| issn | 2079-4991 |
| language | English |
| last_indexed | 2024-03-10T00:49:32Z |
| publishDate | 2022-01-01 |
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| series | Nanomaterials |
| spelling | doaj.art-f8b0f8e7f825414bbf54547d222d81852023-11-23T14:54:51ZengMDPI AGNanomaterials2079-49912022-01-0112222010.3390/nano12020220Assessing Genotoxicity of Ten Different Engineered Nanomaterials by the Novel Semi-Automated FADU Assay and the Alkaline Comet AssaySarah May0Cordula Hirsch1Alexandra Rippl2Alexander Bürkle3Peter Wick4Particles-Biology Interactions Lab, Swiss Federal Laboratories for Materials Science and Technology (EMPA), Lerchenfeldstrasse 5, 9014 St. Gallen, SwitzerlandParticles-Biology Interactions Lab, Swiss Federal Laboratories for Materials Science and Technology (EMPA), Lerchenfeldstrasse 5, 9014 St. Gallen, SwitzerlandParticles-Biology Interactions Lab, Swiss Federal Laboratories for Materials Science and Technology (EMPA), Lerchenfeldstrasse 5, 9014 St. Gallen, SwitzerlandMolecular Toxicology Group, University of Konstanz, Universitätsstrasse 10, 78464 Konstanz, GermanyParticles-Biology Interactions Lab, Swiss Federal Laboratories for Materials Science and Technology (EMPA), Lerchenfeldstrasse 5, 9014 St. Gallen, SwitzerlandIncreased engineered nanomaterial (ENM) production and incorporation in consumer and biomedical products has raised concerns about the potential adverse effects. The DNA damaging capacity is of particular importance since damaged genetic material can lead to carcinogenesis. Consequently, reliable and robust in vitro studies assessing ENM genotoxicity are of great value. We utilized two complementary assays based on different measurement principles: (1) comet assay and (2) FADU (fluorimetric detection of alkaline DNA unwinding) assay. Assessing cell viability ruled out false-positive results due to DNA fragmentation during cell death. Potential structure–activity relationships of 10 ENMs were investigated: three silica nanoparticles (SiO<sub>2</sub>-NP) with varying degrees of porosity, titanium dioxide (TiO<sub>2</sub>-NP), polystyrene (PS-NP), zinc oxide (ZnO-NP), gold (Au-NP), graphene oxide (GO) and two multi-walled carbon nanotubes (MWNT). SiO<sub>2</sub>-NPs, TiO<sub>2</sub>-NP and GO were neither cytotoxic nor genotoxic to Jurkat E6-I cells. Quantitative interference corrections derived from GO results can make the FADU assay a promising screening tool for a variety of ENMs. MWNT merely induced cytotoxicity, while dose- and time-dependent cytotoxicity of PS-NP was accompanied by DNA fragmentation. Hence, PS-NP served to benchmark threshold levels of cytotoxicity at which DNA fragmentation was expected. Considering all controls revealed the true genotoxicity for Au-NP and ZnO-NP at early time points.https://www.mdpi.com/2079-4991/12/2/220comet assayFADU assayengineered nanomaterialsDNA strand breaksgenotoxicityENM interference |
| spellingShingle | Sarah May Cordula Hirsch Alexandra Rippl Alexander Bürkle Peter Wick Assessing Genotoxicity of Ten Different Engineered Nanomaterials by the Novel Semi-Automated FADU Assay and the Alkaline Comet Assay Nanomaterials comet assay FADU assay engineered nanomaterials DNA strand breaks genotoxicity ENM interference |
| title | Assessing Genotoxicity of Ten Different Engineered Nanomaterials by the Novel Semi-Automated FADU Assay and the Alkaline Comet Assay |
| title_full | Assessing Genotoxicity of Ten Different Engineered Nanomaterials by the Novel Semi-Automated FADU Assay and the Alkaline Comet Assay |
| title_fullStr | Assessing Genotoxicity of Ten Different Engineered Nanomaterials by the Novel Semi-Automated FADU Assay and the Alkaline Comet Assay |
| title_full_unstemmed | Assessing Genotoxicity of Ten Different Engineered Nanomaterials by the Novel Semi-Automated FADU Assay and the Alkaline Comet Assay |
| title_short | Assessing Genotoxicity of Ten Different Engineered Nanomaterials by the Novel Semi-Automated FADU Assay and the Alkaline Comet Assay |
| title_sort | assessing genotoxicity of ten different engineered nanomaterials by the novel semi automated fadu assay and the alkaline comet assay |
| topic | comet assay FADU assay engineered nanomaterials DNA strand breaks genotoxicity ENM interference |
| url | https://www.mdpi.com/2079-4991/12/2/220 |
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