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Next-generation sequencing reveals size-dependent pulmonary impact of thin graphene oxide sheets in mice: towards safe-by-design

ABSTRACT: Safety assessment of graphene-based materials (GBMs) including graphene oxide (GO) is essential for their safe use across many sectors of society. In particular, the link between specific material properties and biological effects needs to be further elucidated. Here we compared the effects of lateral dimensions of GO sheets in acute and chronic pulmonary responses after single intranasal instillation in mice. Micrometre-sized GO (1–30 μm) induced stronger pulmonary inflammation than nanometre-sized GO, despite a reduced translocation to the lungs. Genome-wide RNA sequencing also revealed distinct size dependent effects of GO, in agreement with the histopathological results. Although large GO, but not the smallest GO (10–300 nm), triggered the formation of granulomas that persisted up to 90 days, no pulmonary fibrosis was observed. These results could be partly explained by the pattern of size-independent in situ biotransformation of GO, as evidenced by Raman spectroscopy imaging. Our findings demonstrate that lateral dimensions play a fundamental role in the pulmonary response to GO, and suggest that airborne exposure to micrometer sized GO should be avoided in production plant or applications, such as spray coating or painting, where aerosolised dispersions are likely to occur. These results are important for the development of evidence-based risk assessment, and provide information towards the implementation of a safer-by-design approach for GBM enhanced products and applications, for the benefit of workers and end-users.

ORGANISM(S): Mus musculus

PROVIDER: GSE146796 | GEO | 2020/09/30

REPOSITORIES: GEO

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Project description:Graphene-derived materials are a family of nanomaterials with multiple potential applications in different fields such as biomedicine. It is therefore essential to understand their interaction with cellular barriers such as skin. In this work we evaluated the metabolic changes in human skin cells (HaCaT) exposed to different GRMs for 7 and 30 days. Objectives Endogenous metabolic profiles of control and graphene-treated keratinocytes have been studied using ultra-high performance liquid chromatography – mass spectrometry (UHPLC-MS). Keratinocytes were treated with graphene oxide (GO) from two different suppliers and with few layer graphene (FLG). Samples were collected one week and one month after the start of the treatment. The general aim of the project was to evaluate potential metabolic differences between: 1) Graphene-treated keratinocytes and control keratinocytes at one week; 2) Graphene-treated keratinocytes and control keratinocytes at one month; 3) Control keratinocytes at 1 month and 1 week; 4) Graphene-treated keratinocytes at 1 month and 1 week. Experimental Procedures A successful metabolic profiling experiment relies on the ability to determine changes in an organism’s biofluid or tissue complement of metabolites. Mass spectrometry coupled to ultra-high performance liquid chromatography (UHPLC-MS) is well suited to such analyses due to its high sensitivity, large coverage over different classes of metabolites, high throughput capacity, and wide dynamic range. In this study, one UHPLC-MS based platform was used to analyse endogenous analytes for inclusion in subsequent statistical analysis procedures used to study metabolic differences between the groups of samples. Results The oxidation degree and size of the GRMs is determinant in the effect on cell metabolism, as well as the exposure time. Thus, one of the materials used generated a change in the energy metabolism of the cells, significantly increasing the level of different Krebs cycle metabolites.

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Next-generation sequencing reveals size-dependent pulmonary impact of thin graphene oxide sheets in mice: towards safe-by-design

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