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Few-layer graphene induces both primary and secondary genotoxicity in epithelial barrier models in vitro
BACKGROUND: Toxicological evaluation of engineered nanomaterials (ENMs) is essential for occupational health and safety, particularly where bulk manufactured ENMs such as few-layer graphene (FLG) are concerned. Additionally, there is a necessity to develop advanced in vitro models when testing ENMs...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7816456/ https://www.ncbi.nlm.nih.gov/pubmed/33468168 http://dx.doi.org/10.1186/s12951-021-00769-9 |
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author | Burgum, Michael J. Clift, Martin J. D. Evans, Stephen J. Hondow, Nicole Tarat, Afshin Jenkins, Gareth J. Doak, Shareen H. |
author_facet | Burgum, Michael J. Clift, Martin J. D. Evans, Stephen J. Hondow, Nicole Tarat, Afshin Jenkins, Gareth J. Doak, Shareen H. |
author_sort | Burgum, Michael J. |
collection | PubMed |
description | BACKGROUND: Toxicological evaluation of engineered nanomaterials (ENMs) is essential for occupational health and safety, particularly where bulk manufactured ENMs such as few-layer graphene (FLG) are concerned. Additionally, there is a necessity to develop advanced in vitro models when testing ENMs to provide a physiologically relevant alternative to invasive animal experimentation. The aim of this study was to determine the genotoxicity of non-functionalised (neutral), amine- and carboxyl-functionalised FLG upon both human-transformed type-I (TT1) alveolar epithelial cell monocultures, as well as co-cultures of TT1 and differentiated THP-1 monocytes (d.THP-1 (macrophages)). RESULTS: In monocultures, TT1 and d.THP-1 macrophages showed a statistically significant (p < 0.05) cytotoxic response with each ENM following 24-h exposures. Monoculture genotoxicity measured by the in vitro cytokinesis blocked micronucleus (CBMN) assay revealed significant (p < 0.05) micronuclei induction at 8 µg/ml for amine- and carboxyl-FLG. Transmission electron microscopy (TEM) revealed ENMs were internalised by TT1 cells within membrane-bound vesicles. In the co-cultures, ENMs induced genotoxicity in the absence of cytotoxic effects. Co-cultures pre-exposed to 1.5 mM N-acetylcysteine (NAC), showed baseline levels of micronuclei induction, indicating that the genotoxicity observed was driven by oxidative stress. CONCLUSIONS: Therefore, FLG genotoxicity when examined in monocultures, results in primary-indirect DNA damage; whereas co-cultured cells reveal secondary mechanisms of DNA damage. [Image: see text] |
format | Online Article Text |
id | pubmed-7816456 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-78164562021-01-22 Few-layer graphene induces both primary and secondary genotoxicity in epithelial barrier models in vitro Burgum, Michael J. Clift, Martin J. D. Evans, Stephen J. Hondow, Nicole Tarat, Afshin Jenkins, Gareth J. Doak, Shareen H. J Nanobiotechnology Research BACKGROUND: Toxicological evaluation of engineered nanomaterials (ENMs) is essential for occupational health and safety, particularly where bulk manufactured ENMs such as few-layer graphene (FLG) are concerned. Additionally, there is a necessity to develop advanced in vitro models when testing ENMs to provide a physiologically relevant alternative to invasive animal experimentation. The aim of this study was to determine the genotoxicity of non-functionalised (neutral), amine- and carboxyl-functionalised FLG upon both human-transformed type-I (TT1) alveolar epithelial cell monocultures, as well as co-cultures of TT1 and differentiated THP-1 monocytes (d.THP-1 (macrophages)). RESULTS: In monocultures, TT1 and d.THP-1 macrophages showed a statistically significant (p < 0.05) cytotoxic response with each ENM following 24-h exposures. Monoculture genotoxicity measured by the in vitro cytokinesis blocked micronucleus (CBMN) assay revealed significant (p < 0.05) micronuclei induction at 8 µg/ml for amine- and carboxyl-FLG. Transmission electron microscopy (TEM) revealed ENMs were internalised by TT1 cells within membrane-bound vesicles. In the co-cultures, ENMs induced genotoxicity in the absence of cytotoxic effects. Co-cultures pre-exposed to 1.5 mM N-acetylcysteine (NAC), showed baseline levels of micronuclei induction, indicating that the genotoxicity observed was driven by oxidative stress. CONCLUSIONS: Therefore, FLG genotoxicity when examined in monocultures, results in primary-indirect DNA damage; whereas co-cultured cells reveal secondary mechanisms of DNA damage. [Image: see text] BioMed Central 2021-01-19 /pmc/articles/PMC7816456/ /pubmed/33468168 http://dx.doi.org/10.1186/s12951-021-00769-9 Text en © The Author(s) 2021 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Burgum, Michael J. Clift, Martin J. D. Evans, Stephen J. Hondow, Nicole Tarat, Afshin Jenkins, Gareth J. Doak, Shareen H. Few-layer graphene induces both primary and secondary genotoxicity in epithelial barrier models in vitro |
title | Few-layer graphene induces both primary and secondary genotoxicity in epithelial barrier models in vitro |
title_full | Few-layer graphene induces both primary and secondary genotoxicity in epithelial barrier models in vitro |
title_fullStr | Few-layer graphene induces both primary and secondary genotoxicity in epithelial barrier models in vitro |
title_full_unstemmed | Few-layer graphene induces both primary and secondary genotoxicity in epithelial barrier models in vitro |
title_short | Few-layer graphene induces both primary and secondary genotoxicity in epithelial barrier models in vitro |
title_sort | few-layer graphene induces both primary and secondary genotoxicity in epithelial barrier models in vitro |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7816456/ https://www.ncbi.nlm.nih.gov/pubmed/33468168 http://dx.doi.org/10.1186/s12951-021-00769-9 |
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