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Dynamic interactions and intracellular fate of label-free GO within mammalian cells: role of lateral sheet size

Yingxian Chen, Livia Elena Crică, Vinicio Rosano, Adrián Esteban Arranz, David Spiller, Kostas Kostarelos, Sandra Vranic
doi: https://doi.org/10.1101/805200
Yingxian Chen
1Nanomedicine Lab, Faculty of Biology, Medicine and Health, The University of Manchester, AV Hill Building, Manchester M13 9PT, U.K.
2National Graphene Institute, The University of Manchester, Booth Street East, Manchester M13 9PL, U.K.
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Livia Elena Crică
1Nanomedicine Lab, Faculty of Biology, Medicine and Health, The University of Manchester, AV Hill Building, Manchester M13 9PT, U.K.
2National Graphene Institute, The University of Manchester, Booth Street East, Manchester M13 9PL, U.K.
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Vinicio Rosano
1Nanomedicine Lab, Faculty of Biology, Medicine and Health, The University of Manchester, AV Hill Building, Manchester M13 9PT, U.K.
2National Graphene Institute, The University of Manchester, Booth Street East, Manchester M13 9PL, U.K.
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Adrián Esteban Arranz
1Nanomedicine Lab, Faculty of Biology, Medicine and Health, The University of Manchester, AV Hill Building, Manchester M13 9PT, U.K.
2National Graphene Institute, The University of Manchester, Booth Street East, Manchester M13 9PL, U.K.
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David Spiller
3FBMH Platform Sciences, Enabling Technologies & Infrastructure, FBMH Research & Innovation, Faculty of Biology, Medicine and Health, The University of Manchester, Michael Smith Building, Manchester M13 9PT, U.K.
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Kostas Kostarelos
1Nanomedicine Lab, Faculty of Biology, Medicine and Health, The University of Manchester, AV Hill Building, Manchester M13 9PT, U.K.
2National Graphene Institute, The University of Manchester, Booth Street East, Manchester M13 9PL, U.K.
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  • For correspondence: kostas.kostarelos@manchester.ac.uk sandra.vranic@manchester.ac.uk
Sandra Vranic
1Nanomedicine Lab, Faculty of Biology, Medicine and Health, The University of Manchester, AV Hill Building, Manchester M13 9PT, U.K.
2National Graphene Institute, The University of Manchester, Booth Street East, Manchester M13 9PL, U.K.
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  • For correspondence: kostas.kostarelos@manchester.ac.uk sandra.vranic@manchester.ac.uk
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Abstract

Graphene oxide (GO) holds great potential for biomedical applications, however fundamental understanding of the way it interacts with biological systems is still lacking even though it is a prerequisite for successful clinical translation. In this study, we exploited intrinsic fluorescent properties of GO to establish the relationship between lateral dimensions of the material, its cellular uptake mechanism and intracellular fate. Label-free GO with distinct lateral dimensions, small (s-GO) and ultra-small (us-GO), was synthesized and thoroughly characterised both in water and in biologically relevant cell culture medium. Interactions of the material with a range of non-phagocytic mammalian cell lines (BEAS-2B, NIH/3T3, HaCaT, 293T) were studied using a combination of complementary analytical techniques (confocal microscopy, flow cytometry and TEM). The uptake mechanism was interrogated using a range of pharmaceutical inhibitors for main endocytic pathways (ethyl-isopropyl amiloride, monodansylcadaverine, chlorpromazine, genistein, cytochalasin D, latrunculin A, dynasore and sodium azide), and validated using negatively charged polystyrene beads with different diameters (0.1 and 1 μm). Regardless of lateral dimension, both types of GO were found to interact with the plasma membrane and to be efficiently taken up by a panel of cell lines in a time- and dose-dependent manner. s-GO was internalised mainly via macropinocytosis while us-GO used mainly clathrin- and caveolae-mediated endocytosis. Lastly, we show that both s-GO and us-GO terminate in lysosomal compartments for up to 48 h. Our results aim to offer significant insight into the mechanism of interaction of GO with non-phagocytic cell lines that can be exploited for the design of biomedically applicable 2D transport systems.

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Posted October 16, 2019.
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Dynamic interactions and intracellular fate of label-free GO within mammalian cells: role of lateral sheet size
Yingxian Chen, Livia Elena Crică, Vinicio Rosano, Adrián Esteban Arranz, David Spiller, Kostas Kostarelos, Sandra Vranic
bioRxiv 805200; doi: https://doi.org/10.1101/805200
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Dynamic interactions and intracellular fate of label-free GO within mammalian cells: role of lateral sheet size
Yingxian Chen, Livia Elena Crică, Vinicio Rosano, Adrián Esteban Arranz, David Spiller, Kostas Kostarelos, Sandra Vranic
bioRxiv 805200; doi: https://doi.org/10.1101/805200

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