Intracellular localization and toxicity of graphene oxide and reduced graphene oxide nanoplatelets to mussel hemocytes in vitro
[Display omitted] •PVP increases the stability, bioavailability and toxicity of GO and rGO nanoplatelets.•Physico-chemical properties of GO such as lateral dimension and oxygen content influence their toxicity.•GO and rGO-PVP disrupt the cell membrane integrity and their toxicity is ROS-mediated.•Bo...
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Published in | Aquatic toxicology Vol. 188; pp. 138 - 147 |
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Main Authors | , , |
Format | Journal Article |
Language | English |
Published |
Netherlands
Elsevier B.V
01.07.2017
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Subjects | |
Online Access | Get full text |
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Abstract | [Display omitted]
•PVP increases the stability, bioavailability and toxicity of GO and rGO nanoplatelets.•Physico-chemical properties of GO such as lateral dimension and oxygen content influence their toxicity.•GO and rGO-PVP disrupt the cell membrane integrity and their toxicity is ROS-mediated.•Both GO and rGO are internalized in mussel hemocytes being accumulation of the latter more evident.
Recently, graphene materials have attracted tremendous research interest due to their unique physicochemical properties that hold great promise in electronics, energy, materials and biomedical areas. Graphene oxide (GO) is one of the most extensively studied graphene derivatives. In order to improve GO electrical properties, nanoplatelets are chemically reduced, thus increasing nanoplatelet conductivity. This reduced GO (rGO) shows different properties and behavior compared to GO. Graphene-based wastes are expected to end up in the marine environment. Here we aimed to assess the potential toxic effects of GO and rGO to marine organisms by using in vitro assays with mussel (Mytilus galloprovincialis) hemocytes. Cells were exposed to a wide range of concentrations (up to 100mg/L) of GO (with and without polyvinylpyrrolidone-PVP as stabilizing agent: GO and GO-PVP) and rGO with PVP (rGO-PVP) to assess cytotoxicity and cell membrane integrity. Then, cells were exposed to sublethal concentrations of GO and rGO-PVP to assess their subcellular distribution through transmission electron microscopy (TEM) and to evaluate their effects on ROS production. GO, GO-PVP and rGO-PVP showed low and concentration-dependent cytotoxicity. rGO-PVP (LC50=29.902 and 33.94mg/L depending on the origin) was more toxic than GO (LC50=49.84 and 54.51mg/L depending on the origin) and GO-PVP (LC50=43.72mg/L). PVP was not toxic to hemocytes but increased bioavailability and toxicity of nanoplatelets. At TEM, GO and rGO-PVP nanoplatelets caused invaginations and perforations of the plasma membrane, which agrees with the observed decrease in cell membrane integrity. Nanoplatelets were internalized, at a higher extent for rGO-PVP than for GO, and found in the cytosol and in endolysosomal vesicles of hemocytes. Both GO and rGO-PVP increased ROS production at the highest sublethal concentration tested. In conclusion, GO, GO-PVP and rGO-PVP are not highly toxic to mussel cells but they cause membrane damage and their toxicity is ROS-mediated. Finally, in vitro assays with mussel hemocytes are sensitive tools to detect toxic effects of graphene-based nanomaterials. |
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AbstractList | [Display omitted]
•PVP increases the stability, bioavailability and toxicity of GO and rGO nanoplatelets.•Physico-chemical properties of GO such as lateral dimension and oxygen content influence their toxicity.•GO and rGO-PVP disrupt the cell membrane integrity and their toxicity is ROS-mediated.•Both GO and rGO are internalized in mussel hemocytes being accumulation of the latter more evident.
Recently, graphene materials have attracted tremendous research interest due to their unique physicochemical properties that hold great promise in electronics, energy, materials and biomedical areas. Graphene oxide (GO) is one of the most extensively studied graphene derivatives. In order to improve GO electrical properties, nanoplatelets are chemically reduced, thus increasing nanoplatelet conductivity. This reduced GO (rGO) shows different properties and behavior compared to GO. Graphene-based wastes are expected to end up in the marine environment. Here we aimed to assess the potential toxic effects of GO and rGO to marine organisms by using in vitro assays with mussel (Mytilus galloprovincialis) hemocytes. Cells were exposed to a wide range of concentrations (up to 100mg/L) of GO (with and without polyvinylpyrrolidone-PVP as stabilizing agent: GO and GO-PVP) and rGO with PVP (rGO-PVP) to assess cytotoxicity and cell membrane integrity. Then, cells were exposed to sublethal concentrations of GO and rGO-PVP to assess their subcellular distribution through transmission electron microscopy (TEM) and to evaluate their effects on ROS production. GO, GO-PVP and rGO-PVP showed low and concentration-dependent cytotoxicity. rGO-PVP (LC50=29.902 and 33.94mg/L depending on the origin) was more toxic than GO (LC50=49.84 and 54.51mg/L depending on the origin) and GO-PVP (LC50=43.72mg/L). PVP was not toxic to hemocytes but increased bioavailability and toxicity of nanoplatelets. At TEM, GO and rGO-PVP nanoplatelets caused invaginations and perforations of the plasma membrane, which agrees with the observed decrease in cell membrane integrity. Nanoplatelets were internalized, at a higher extent for rGO-PVP than for GO, and found in the cytosol and in endolysosomal vesicles of hemocytes. Both GO and rGO-PVP increased ROS production at the highest sublethal concentration tested. In conclusion, GO, GO-PVP and rGO-PVP are not highly toxic to mussel cells but they cause membrane damage and their toxicity is ROS-mediated. Finally, in vitro assays with mussel hemocytes are sensitive tools to detect toxic effects of graphene-based nanomaterials. Recently, graphene materials have attracted tremendous research interest due to their unique physicochemical properties that hold great promise in electronics, energy, materials and biomedical areas. Graphene oxide (GO) is one of the most extensively studied graphene derivatives. In order to improve GO electrical properties, nanoplatelets are chemically reduced, thus increasing nanoplatelet conductivity. This reduced GO (rGO) shows different properties and behavior compared to GO. Graphene-based wastes are expected to end up in the marine environment. Here we aimed to assess the potential toxic effects of GO and rGO to marine organisms by using in vitro assays with mussel (Mytilus galloprovincialis) hemocytes. Cells were exposed to a wide range of concentrations (up to 100mg/L) of GO (with and without polyvinylpyrrolidone-PVP as stabilizing agent: GO and GO-PVP) and rGO with PVP (rGO-PVP) to assess cytotoxicity and cell membrane integrity. Then, cells were exposed to sublethal concentrations of GO and rGO-PVP to assess their subcellular distribution through transmission electron microscopy (TEM) and to evaluate their effects on ROS production. GO, GO-PVP and rGO-PVP showed low and concentration-dependent cytotoxicity. rGO-PVP (LC50=29.902 and 33.94mg/L depending on the origin) was more toxic than GO (LC50=49.84 and 54.51mg/L depending on the origin) and GO-PVP (LC50=43.72mg/L). PVP was not toxic to hemocytes but increased bioavailability and toxicity of nanoplatelets. At TEM, GO and rGO-PVP nanoplatelets caused invaginations and perforations of the plasma membrane, which agrees with the observed decrease in cell membrane integrity. Nanoplatelets were internalized, at a higher extent for rGO-PVP than for GO, and found in the cytosol and in endolysosomal vesicles of hemocytes. Both GO and rGO-PVP increased ROS production at the highest sublethal concentration tested. In conclusion, GO, GO-PVP and rGO-PVP are not highly toxic to mussel cells but they cause membrane damage and their toxicity is ROS-mediated. Finally, in vitro assays with mussel hemocytes are sensitive tools to detect toxic effects of graphene-based nanomaterials. |
Author | Cajaraville, Miren P. Tomovska, Radmila Katsumiti, Alberto |
Author_xml | – sequence: 1 givenname: Alberto surname: Katsumiti fullname: Katsumiti, Alberto organization: CBET Research Group, Dept. Zoology and Animal Cell Biology, Faculty of Science and Technology and Research Centre for Experimental Marine Biology and Biotechnology PIE, University of the Basque Country UPV/EHU, Basque Country, Spain – sequence: 2 givenname: Radmila surname: Tomovska fullname: Tomovska, Radmila organization: POLYMAT and Dept. Applied Chemistry, Faculty of Chemistry, University of the Basque Country UPV/EHU, Basque Country, Spain – sequence: 3 givenname: Miren P. surname: Cajaraville fullname: Cajaraville, Miren P. email: mirenp.cajaraville@ehu.eus organization: CBET Research Group, Dept. Zoology and Animal Cell Biology, Faculty of Science and Technology and Research Centre for Experimental Marine Biology and Biotechnology PIE, University of the Basque Country UPV/EHU, Basque Country, Spain |
BackLink | https://www.ncbi.nlm.nih.gov/pubmed/28521151$$D View this record in MEDLINE/PubMed |
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Keywords | Cytotoxicity Mussel hemocytes Intracellular localization Graphene oxide (GO) and reduced GO Membrane damage ROS production |
Language | English |
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7 Katsumiti (10.1016/j.aquatox.2017.04.016_bib0125) 2015; 10 Li (10.1016/j.aquatox.2017.04.016_bib0145) 2012; 33 Orbea (10.1016/j.aquatox.2017.04.016_bib0200) 2006; 25 Borja (10.1016/j.aquatox.2017.04.016_bib0030) 2004 Lammel (10.1016/j.aquatox.2017.04.016_bib0140) 2013; 10 Seabra (10.1016/j.aquatox.2017.04.016_bib0235) 2014; 27 Moore (10.1016/j.aquatox.2017.04.016_bib0185) 2006; 32 |
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•PVP increases the stability, bioavailability and toxicity of GO and rGO nanoplatelets.•Physico-chemical properties of GO such as lateral... Recently, graphene materials have attracted tremendous research interest due to their unique physicochemical properties that hold great promise in electronics,... |
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SubjectTerms | Animals Cell Survival - drug effects Cytosol - metabolism Cytotoxicity Graphene oxide (GO) and reduced GO Graphite - toxicity Hemocytes - drug effects Hemocytes - metabolism Intracellular localization Membrane damage Microscopy, Electron, Transmission Mussel hemocytes Mytilus Nanostructures - toxicity Nanostructures - ultrastructure Oxides - toxicity ROS production Water Pollutants, Chemical - toxicity |
Title | Intracellular localization and toxicity of graphene oxide and reduced graphene oxide nanoplatelets to mussel hemocytes in vitro |
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