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Aquat Toxicol. 2014 May;150:55-65. doi: 10.1016/j.aquatox.2014.02.016. Epub 2014 Mar 2.

Graphene nanoplatelets spontaneously translocate into the cytosol and physically interact with cellular organelles in the fish cell line PLHC-1.

Author information

1
Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Departamento de Medio Ambiente, Carretera de la Coruña Km 7.5, Madrid 28040, Spain.
2
Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Departamento de Medio Ambiente, Carretera de la Coruña Km 7.5, Madrid 28040, Spain. Electronic address: jmnavas@inia.es.

Abstract

Graphene and graphene derivatives constitute a novel class of carbon-based nanomaterials being increasingly produced and used in technical and consumer applications. Release of graphene nanoplatelets during the life cycle of these applications may result in human and environmental exposure calling for assessment of their potential to cause harm to humans and wildlife. This study aimed to assess the toxicity of graphene oxide (GO) and carboxyl graphene (CXYG) nanoplatelets to non-mammalian species using the fish cell line PLHC-1 as in vitro model. The cytotoxicity of GO and CXYG was assessed using different assays measuring alterations in plasma membrane integrity, metabolic activity, and lysosomal and mitochondrial function. The induction of oxidative stress was assessed by measuring intracellular reactive oxygen species (ROS) levels. Interaction with the plasma membrane and internalization of nanoplatelets were investigated by electron microscopy. Graphene nanoplatelets spontaneously penetrated through the plasma membrane and accumulated in the cytosol, where they further interacted with mitochondrial and nuclear membranes. PLHC-1 cells demonstrated significantly reduced mitochondrial membrane potential (MMP) and increased ROS levels at 16 μg/ml GO and CXYG (72 h), but barely any decrease in cell viability. The observation of intracellular graphene accumulations not enclosed by membranes suggests that GO and CXYG internalization in fish hepatoma cells occurs through an endocytosis-independent mechanism.

KEYWORDS:

Carbon nanomaterial; Cytotoxicity; Graphene oxide; In vitro; Mitochondria; Reactive oxygen species

PMID:
24642293
DOI:
10.1016/j.aquatox.2014.02.016
[Indexed for MEDLINE]

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