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Adapting the in vitro micronucleus assay (OECD Test Guideline No. 487) for testing of manufactured nanomaterials: recommendations for best practices
,
Caroline Gomes,
Svenja Berit Seiffert,
Robert Landsiedel,
Naveed Honarvar,
Shareen Doak
Mutagenesis, Volume: 39, Issue: 3, Pages: 205 - 217
Swansea University Authors:
Michael Burgum, Stephen Evans , Shareen Doak
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DOI (Published version): 10.1093/mutage/geae010
Abstract
The current Organisation for Economic Co-Operation and Development test guideline number 487 (OECD TG No. 487) provides instruction on how to conduct the in vitro micronucleus assay. This assay is one of the gold standard approaches for measuring the mutagenicity of test items; however, it is direct...
| Published in: | Mutagenesis |
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| ISSN: | 0267-8357 1464-3804 |
| Published: |
Oxford University Press (OUP)
2024
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| Online Access: |
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| URI: | https://cronfa.swan.ac.uk/Record/cronfa65963 |
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| Abstract: |
The current Organisation for Economic Co-Operation and Development test guideline number 487 (OECD TG No. 487) provides instruction on how to conduct the in vitro micronucleus assay. This assay is one of the gold standard approaches for measuring the mutagenicity of test items; however, it is directed at testing low molecular weight molecules and may not be appropriate for particulate materials (e.g. engineered nanoparticles [ENPs]). This study aimed to adapt the in vitro micronucleus assay for ENP testing and underpins the development of an OECD guidance document. A harmonized, nano-specific protocol was generated and evaluated by two independent laboratories. Cell lines utilized were human lymphoblastoid (TK6) cells, human liver hepatocytes (HepG2) cells, Chinese hamster lung fibroblast (V79) cells, whole blood, and buffy coat cells from healthy human volunteers. These cells were exposed to reference ENPs from the Joint Research Council (JRC): SiO2 (RLS-0102), Au5nm and Au30nm (RLS-03, RLS-010), CeO2 (NM212), and BaSO4 (NM220). Tungsten carbide-cobalt (WC/Co) was used as a trial particulate positive control. The chemical controls were positive in all cell cultures, but WC/Co was only positive in TK6 and buffy coat cells. In TK6 cells, mutagenicity was observed for SiO2- and both Au types. In HepG2 cells, Au5nm and SiO2 showed sub-two-fold increases in micronuclei. In V79 cells, whole blood, and buffy coat cells, no genotoxicity was detected with the test materials. The data confirmed that ENPs could be tested with the harmonized protocol, additionally, concordant data were observed across the two laboratories with V79 cells. WC/Co may be a suitable particulate positive control in the in vitro micronucleus assay when using TK6 and buffy coat cells. Detailed recommendations are therefore provided to adapt OECD TG No. 487 for testing ENP. |
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| Keywords: |
Nanoparticles, micronucleus, WC/Co, OECD |
| College: |
Faculty of Medicine, Health and Life Sciences |
| Funders: |
The authors would like to acknowledge this research has received funding from the European Union’s Horizon 2020 research and innovation program for the RiskGONE project, grant agreement 814425. S.J.E. was supported by the Celtic Advanced Life Science Innovation Network, an Ireland-Wales 2014-2020 programme part funded by the European Regional Development Fund through the Welsh Government (80885). The data generated in this manuscript are available from the corresponding author upon reasonable request. |
| Issue: |
3 |
| Start Page: |
205 |
| End Page: |
217 |