Screening the micronucleus assay for reliable estimation of the genotoxicity of graphene and other 2D materials

In recent years, the genotoxicity of graphene-related materials (GRMs) and other 2D materials has been evaluated using different models to ensure their safety. The OECD TG 487 in vitro micronucleus (MNu) test is one of the most widely used methods to assess the genotoxicity of nanomaterials (NMs). H...

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Autores: Esteban Arranz, Adrián, García-Carpintero Fernández-Pacheco, Sonia, González Velázquez, Viviana Jehova, Frontiñan Rubio, Javier, Durán Prado, Mario, Vázquez Fernández-Pacheco, Ester
Formato: artículo
Fecha de publicación:2023
País:España
Recursos:Universidad de Castilla-La Mancha
Repositorio:RUIdeRA. Repositorio Institucional de la UCLM
OAI Identifier:oai:ruidera.uclm.es:10578/32463
Acesso em linha:https://www.sciencedirect.com/science/article/pii/S0008622323006711
https://hdl.handle.net/10578/32463
Access Level:acceso abierto
Palavra-chave:2D nanomaterials
Ensayo de micronúcleos
Genotoxicidad
Genotoxicity
Grafeno
Graphene
Micronucleus assay
Nanomateriales 2D
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spelling Screening the micronucleus assay for reliable estimation of the genotoxicity of graphene and other 2D materialsEsteban Arranz, AdriánGarcía-Carpintero Fernández-Pacheco, SoniaGonzález Velázquez, Viviana JehovaFrontiñan Rubio, JavierDurán Prado, MarioVázquez Fernández-Pacheco, Ester2D nanomaterialsEnsayo de micronúcleosGenotoxicidadGenotoxicityGrafenoGrapheneMicronucleus assayNanomateriales 2DIn recent years, the genotoxicity of graphene-related materials (GRMs) and other 2D materials has been evaluated using different models to ensure their safety. The OECD TG 487 in vitro micronucleus (MNu) test is one of the most widely used methods to assess the genotoxicity of nanomaterials (NMs). However, the nature of NMs interferes with the standard MNu process, thus requiring an adaptation to assess this kind of compound. Improvements have been incorporated into the standard method to solve the interferences and build a modified, reliable protocol that can be used to test NMs further. The main interference observed was NM agglomeration, which hinders the correct visualization of micronuclei. Addition of 10% dextran to the culture medium reduced agglomeration but interfered with toxicity by reducing NM-induced cell death. Mechanical shaking during cell incubation with NMs minimized the size and number of agglomerates and avoided their rapid sedimentation and deposition onto cells, thus allowing accurate visualization of the micronucleate cells. Once set up, the proposed method was tested with graphene oxide (GO), few-layer graphene (FLG), molybdenum disulfide (MoS2), and boron nitride (hBN). The novel in vitro MNu test for NMs showed that GO (0.05–5 µg/ml) and hBN (0.05 µg/ml) were genotoxic at sublethal doses, inducing irreparable chromosomal damage. With these adaptations, the modified in vitro MNu test is strongly recommended to test the genotoxicity of NMs. The results obtained with this protocol indicate that GO and hBN represent a risk that should be considered.En los últimos años, la genotoxicidad de los materiales relacionados con el grafeno (GRM) y otros materiales 2D se ha evaluado utilizando diferentes modelos para garantizar su seguridad. El ensayo de micronúcleos in vitro (MNu) OCDE TG 487 es uno de los métodos más utilizados para evaluar la genotoxicidad de los nanomateriales (NMs). Sin embargo, la naturaleza de los NM interfiere con el proceso estándar de MNu, por lo que es necesaria una adaptación para evaluar este tipo de compuestos. Se han incorporado mejoras en el método estándar para resolver las interferencias y construir un protocolo modificado y fiable que pueda utilizarse para seguir ensayando NMs. La principal interferencia observada fue la aglomeración de NM, que dificulta la correcta visualización de los micronúcleos. La adición de un 10% de dextrano al medio de cultivo redujo la aglomeración, pero interfirió con la toxicidad al reducir la muerte celular inducida por NM. La agitación mecánica durante la incubación de las células con NMs minimizó el tamaño y número de aglomerados y evitó su rápida sedimentación y deposición sobre las células, permitiendo así una visualización precisa de los micronúcleos. Una vez configurado, el método propuesto se probó con óxido de grafeno (GO), grafeno de pocas capas (FLG), disulfuro de molibdeno (MoS2) y nitruro de boro (hBN). El novedoso ensayo in vitro MNu para NMs demostró que el GO (0,05-5 µg/ml) y el hBN (0,05 µg/ml) eran genotóxicos a dosis subletales, induciendo daños cromosómicos irreparables. Con estas adaptaciones, se recomienda encarecidamente el ensayo in vitro modificado de MNu para probar la genotoxicidad de los NM. Los resultadosElsevier202320232023info:eu-repo/semantics/articleapplication/pdfapplication/pdfhttps://www.sciencedirect.com/science/article/pii/S0008622323006711https://hdl.handle.net/10578/32463reponame:RUIdeRA. Repositorio Institucional de la UCLMinstname:Universidad de Castilla-La ManchaInglésinfo:eu-repo/semantics/openAccessoai:ruidera.uclm.es:10578/324632026-05-27T07:36:41Z
dc.title.none.fl_str_mv Screening the micronucleus assay for reliable estimation of the genotoxicity of graphene and other 2D materials
title Screening the micronucleus assay for reliable estimation of the genotoxicity of graphene and other 2D materials
spellingShingle Screening the micronucleus assay for reliable estimation of the genotoxicity of graphene and other 2D materials
Esteban Arranz, Adrián
2D nanomaterials
Ensayo de micronúcleos
Genotoxicidad
Genotoxicity
Grafeno
Graphene
Micronucleus assay
Nanomateriales 2D
title_short Screening the micronucleus assay for reliable estimation of the genotoxicity of graphene and other 2D materials
title_full Screening the micronucleus assay for reliable estimation of the genotoxicity of graphene and other 2D materials
title_fullStr Screening the micronucleus assay for reliable estimation of the genotoxicity of graphene and other 2D materials
title_full_unstemmed Screening the micronucleus assay for reliable estimation of the genotoxicity of graphene and other 2D materials
title_sort Screening the micronucleus assay for reliable estimation of the genotoxicity of graphene and other 2D materials
dc.creator.none.fl_str_mv Esteban Arranz, Adrián
García-Carpintero Fernández-Pacheco, Sonia
González Velázquez, Viviana Jehova
Frontiñan Rubio, Javier
Durán Prado, Mario
Vázquez Fernández-Pacheco, Ester
author Esteban Arranz, Adrián
author_facet Esteban Arranz, Adrián
García-Carpintero Fernández-Pacheco, Sonia
González Velázquez, Viviana Jehova
Frontiñan Rubio, Javier
Durán Prado, Mario
Vázquez Fernández-Pacheco, Ester
author_role author
author2 García-Carpintero Fernández-Pacheco, Sonia
González Velázquez, Viviana Jehova
Frontiñan Rubio, Javier
Durán Prado, Mario
Vázquez Fernández-Pacheco, Ester
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv 2D nanomaterials
Ensayo de micronúcleos
Genotoxicidad
Genotoxicity
Grafeno
Graphene
Micronucleus assay
Nanomateriales 2D
topic 2D nanomaterials
Ensayo de micronúcleos
Genotoxicidad
Genotoxicity
Grafeno
Graphene
Micronucleus assay
Nanomateriales 2D
description In recent years, the genotoxicity of graphene-related materials (GRMs) and other 2D materials has been evaluated using different models to ensure their safety. The OECD TG 487 in vitro micronucleus (MNu) test is one of the most widely used methods to assess the genotoxicity of nanomaterials (NMs). However, the nature of NMs interferes with the standard MNu process, thus requiring an adaptation to assess this kind of compound. Improvements have been incorporated into the standard method to solve the interferences and build a modified, reliable protocol that can be used to test NMs further. The main interference observed was NM agglomeration, which hinders the correct visualization of micronuclei. Addition of 10% dextran to the culture medium reduced agglomeration but interfered with toxicity by reducing NM-induced cell death. Mechanical shaking during cell incubation with NMs minimized the size and number of agglomerates and avoided their rapid sedimentation and deposition onto cells, thus allowing accurate visualization of the micronucleate cells. Once set up, the proposed method was tested with graphene oxide (GO), few-layer graphene (FLG), molybdenum disulfide (MoS2), and boron nitride (hBN). The novel in vitro MNu test for NMs showed that GO (0.05–5 µg/ml) and hBN (0.05 µg/ml) were genotoxic at sublethal doses, inducing irreparable chromosomal damage. With these adaptations, the modified in vitro MNu test is strongly recommended to test the genotoxicity of NMs. The results obtained with this protocol indicate that GO and hBN represent a risk that should be considered.
publishDate 2023
dc.date.none.fl_str_mv 2023
2023
2023
dc.type.none.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv https://www.sciencedirect.com/science/article/pii/S0008622323006711
https://hdl.handle.net/10578/32463
url https://www.sciencedirect.com/science/article/pii/S0008622323006711
https://hdl.handle.net/10578/32463
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv reponame:RUIdeRA. Repositorio Institucional de la UCLM
instname:Universidad de Castilla-La Mancha
instname_str Universidad de Castilla-La Mancha
reponame_str RUIdeRA. Repositorio Institucional de la UCLM
collection RUIdeRA. Repositorio Institucional de la UCLM
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15.300719

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