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Toxicity of Carbon Nanomaterials—Towards Reliable Viability Assessment via New Approach in Flow Cytometry

The scope of application of carbon nanomaterials in biomedical, environmental and industrial fields is recently substantially increasing. Since in vitro toxicity testing is the first essential step for any commercial usage, it is crucial to have a reliable method to analyze the potentially harmful e...

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Detalles Bibliográficos
Autores principales: Malina, Tomáš, Poláková, Kateřina, Hirsch, Cordula, Svoboda, Ladislav, Zbořil, Radek
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8305450/
https://www.ncbi.nlm.nih.gov/pubmed/34299367
http://dx.doi.org/10.3390/ijms22147750
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author Malina, Tomáš
Poláková, Kateřina
Hirsch, Cordula
Svoboda, Ladislav
Zbořil, Radek
author_facet Malina, Tomáš
Poláková, Kateřina
Hirsch, Cordula
Svoboda, Ladislav
Zbořil, Radek
author_sort Malina, Tomáš
collection PubMed
description The scope of application of carbon nanomaterials in biomedical, environmental and industrial fields is recently substantially increasing. Since in vitro toxicity testing is the first essential step for any commercial usage, it is crucial to have a reliable method to analyze the potentially harmful effects of carbon nanomaterials. Even though researchers already reported the interference of carbon nanomaterials with common toxicity assays, there is still, unfortunately, a large number of studies that neglect this fact. In this study, we investigated interference of four bio-promising carbon nanomaterials (graphene acid (GA), cyanographene (GCN), graphitic carbon nitride (g-C(3)N(4)) and carbon dots (QCDs)) in commonly used LIVE/DEAD assay. When a standard procedure was applied, materials caused various types of interference. While positively charged g-C(3)N(4) and QCDs induced false results through the creation of free agglomerates and intrinsic fluorescence properties, negatively charged GA and GCN led to false signals due to the complex quenching effect of the fluorescent dye of a LIVE/DEAD kit. Thus, we developed a new approach using a specific gating strategy based on additional controls that successfully overcame all types of interference and lead to reliable results in LIVE/DEAD assay. We suggest that the newly developed procedure should be a mandatory tool for all in vitro flow cytometry assays of any class of carbon nanomaterials.
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spelling pubmed-83054502021-07-25 Toxicity of Carbon Nanomaterials—Towards Reliable Viability Assessment via New Approach in Flow Cytometry Malina, Tomáš Poláková, Kateřina Hirsch, Cordula Svoboda, Ladislav Zbořil, Radek Int J Mol Sci Article The scope of application of carbon nanomaterials in biomedical, environmental and industrial fields is recently substantially increasing. Since in vitro toxicity testing is the first essential step for any commercial usage, it is crucial to have a reliable method to analyze the potentially harmful effects of carbon nanomaterials. Even though researchers already reported the interference of carbon nanomaterials with common toxicity assays, there is still, unfortunately, a large number of studies that neglect this fact. In this study, we investigated interference of four bio-promising carbon nanomaterials (graphene acid (GA), cyanographene (GCN), graphitic carbon nitride (g-C(3)N(4)) and carbon dots (QCDs)) in commonly used LIVE/DEAD assay. When a standard procedure was applied, materials caused various types of interference. While positively charged g-C(3)N(4) and QCDs induced false results through the creation of free agglomerates and intrinsic fluorescence properties, negatively charged GA and GCN led to false signals due to the complex quenching effect of the fluorescent dye of a LIVE/DEAD kit. Thus, we developed a new approach using a specific gating strategy based on additional controls that successfully overcame all types of interference and lead to reliable results in LIVE/DEAD assay. We suggest that the newly developed procedure should be a mandatory tool for all in vitro flow cytometry assays of any class of carbon nanomaterials. MDPI 2021-07-20 /pmc/articles/PMC8305450/ /pubmed/34299367 http://dx.doi.org/10.3390/ijms22147750 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Malina, Tomáš
Poláková, Kateřina
Hirsch, Cordula
Svoboda, Ladislav
Zbořil, Radek
Toxicity of Carbon Nanomaterials—Towards Reliable Viability Assessment via New Approach in Flow Cytometry
title Toxicity of Carbon Nanomaterials—Towards Reliable Viability Assessment via New Approach in Flow Cytometry
title_full Toxicity of Carbon Nanomaterials—Towards Reliable Viability Assessment via New Approach in Flow Cytometry
title_fullStr Toxicity of Carbon Nanomaterials—Towards Reliable Viability Assessment via New Approach in Flow Cytometry
title_full_unstemmed Toxicity of Carbon Nanomaterials—Towards Reliable Viability Assessment via New Approach in Flow Cytometry
title_short Toxicity of Carbon Nanomaterials—Towards Reliable Viability Assessment via New Approach in Flow Cytometry
title_sort toxicity of carbon nanomaterials—towards reliable viability assessment via new approach in flow cytometry
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8305450/
https://www.ncbi.nlm.nih.gov/pubmed/34299367
http://dx.doi.org/10.3390/ijms22147750
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AT hirschcordula toxicityofcarbonnanomaterialstowardsreliableviabilityassessmentvianewapproachinflowcytometry
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