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Adapting the in vitro micronucleus assay (OECD Test Guideline No. 487) for testing of manufactured nanomaterials: recommendations for best practices

Michael Burgum, Clarissa Ulrich, Natascha Partosa, Stephen Evans Orcid Logo, Caroline Gomes, Svenja Berit Seiffert, Robert Landsiedel, Naveed Honarvar, Shareen Doak Orcid Logo

Mutagenesis, Volume: 39, Issue: 3, Pages: 205 - 217

Swansea University Authors: Michael Burgum, Stephen Evans Orcid Logo, Shareen Doak Orcid Logo

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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...

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Published in: Mutagenesis
ISSN: 0267-8357 1464-3804
Published: Oxford University Press (OUP) 2024
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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. 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spelling v2 65963 2024-04-04 Adapting the in vitro micronucleus assay (OECD Test Guideline No. 487) for testing of manufactured nanomaterials: recommendations for best practices d3fe156a5ee169e586b8bad6ae4cb1d8 Michael Burgum Michael Burgum true false cfca981bdfb8492873a48cc1629def9a 0000-0002-5352-9800 Stephen Evans Stephen Evans true false 8f70286908f67238a527a98cbf66d387 0000-0002-6753-1987 Shareen Doak Shareen Doak true false 2024-04-04 MEDS 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. Journal Article Mutagenesis 39 3 205 217 Oxford University Press (OUP) 0267-8357 1464-3804 Nanoparticles, micronucleus, WC/Co, OECD 24 4 2024 2024-04-24 10.1093/mutage/geae010 http://dx.doi.org/10.1093/mutage/geae010 COLLEGE NANME Medical School COLLEGE CODE MEDS Swansea University SU Library paid the OA fee (TA Institutional Deal) 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. 2024-05-14T16:19:48.2110081 2024-04-04T10:49:03.4153276 Faculty of Medicine, Health and Life Sciences Swansea University Medical School - Biomedical Science Michael Burgum 1 Clarissa Ulrich 2 Natascha Partosa 3 Stephen Evans 0000-0002-5352-9800 4 Caroline Gomes 5 Svenja Berit Seiffert 6 Robert Landsiedel 7 Naveed Honarvar 8 Shareen Doak 0000-0002-6753-1987 9 65963__30368__0c7dc8ee162e43849e5b57c5860f3c0d.pdf 65963.VOR.pdf 2024-05-14T16:06:26.9838042 Output 7150499 application/pdf Version of Record true Distributed under the terms of a Creative Commons Attribution CC-BY Licence. true eng https://creativecommons.org/licenses/by/4.0/
title Adapting the in vitro micronucleus assay (OECD Test Guideline No. 487) for testing of manufactured nanomaterials: recommendations for best practices
spellingShingle Adapting the in vitro micronucleus assay (OECD Test Guideline No. 487) for testing of manufactured nanomaterials: recommendations for best practices
Michael Burgum
Stephen Evans
Shareen Doak
title_short Adapting the in vitro micronucleus assay (OECD Test Guideline No. 487) for testing of manufactured nanomaterials: recommendations for best practices
title_full Adapting the in vitro micronucleus assay (OECD Test Guideline No. 487) for testing of manufactured nanomaterials: recommendations for best practices
title_fullStr Adapting the in vitro micronucleus assay (OECD Test Guideline No. 487) for testing of manufactured nanomaterials: recommendations for best practices
title_full_unstemmed Adapting the in vitro micronucleus assay (OECD Test Guideline No. 487) for testing of manufactured nanomaterials: recommendations for best practices
title_sort Adapting the in vitro micronucleus assay (OECD Test Guideline No. 487) for testing of manufactured nanomaterials: recommendations for best practices
author_id_str_mv d3fe156a5ee169e586b8bad6ae4cb1d8
cfca981bdfb8492873a48cc1629def9a
8f70286908f67238a527a98cbf66d387
author_id_fullname_str_mv d3fe156a5ee169e586b8bad6ae4cb1d8_***_Michael Burgum
cfca981bdfb8492873a48cc1629def9a_***_Stephen Evans
8f70286908f67238a527a98cbf66d387_***_Shareen Doak
author Michael Burgum
Stephen Evans
Shareen Doak
author2 Michael Burgum
Clarissa Ulrich
Natascha Partosa
Stephen Evans
Caroline Gomes
Svenja Berit Seiffert
Robert Landsiedel
Naveed Honarvar
Shareen Doak
format Journal article
container_title Mutagenesis
container_volume 39
container_issue 3
container_start_page 205
publishDate 2024
institution Swansea University
issn 0267-8357
1464-3804
doi_str_mv 10.1093/mutage/geae010
publisher Oxford University Press (OUP)
college_str Faculty of Medicine, Health and Life Sciences
hierarchytype
hierarchy_top_id facultyofmedicinehealthandlifesciences
hierarchy_top_title Faculty of Medicine, Health and Life Sciences
hierarchy_parent_id facultyofmedicinehealthandlifesciences
hierarchy_parent_title Faculty of Medicine, Health and Life Sciences
department_str Swansea University Medical School - Biomedical Science{{{_:::_}}}Faculty of Medicine, Health and Life Sciences{{{_:::_}}}Swansea University Medical School - Biomedical Science
url http://dx.doi.org/10.1093/mutage/geae010
document_store_str 1
active_str 0
description 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.
published_date 2024-04-24T16:19:46Z
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