Gene expression profiling of human macrophages after graphene oxide and graphene nanoplatelets treatment reveals particle-specific regulation of pathways.

Graphene and its derivatives are attractive materials envisaged to enable a wealth of novel applications in many fields including energy, electronics, composite materials or health. A comprehensive understanding of the potential adverse effects of graphene-related materials (GRM) in humans is a prerequisite to the safe use of these promising materials. Here, we exploited gene expression profiling to identify transcriptional responses and toxicity pathways induced by graphene oxide (GO) and graphene nanoplatelets (GNP) in human macrophages. Primary human monocyte-derived macrophages (MDM) and a human macrophage cell line, i.e. differentiated THP-1 cells, were exposed to 5 or 20 µg/mL GO and GNP for 6 and 24 h to capture early and more persistent acute responses at realistic or slightly overdose concentrations. GO and GNP induced time-, dose- and macrophage type-specific differential expression of a substantial number of genes with some overlap between the two GRM types (up to 384 genes (9.6%) or 447 genes (20.4%) in THP-1 or MDM, respectively) but also a high number of genes exclusively deregulated from each material type. Furthermore, GRM responses on gene expression were highly different from those induced by inflammogenic material crystalline quartz (maximum of 64 (2.3%) or 318 (11.3%) common genes for MDM treated with 20 µg/mL GO and GNP, respectively). Further bioinformatics analysis revealed that GNP predominantly activated genes controlling inflammatory and apoptotic pathways whereas GO showed only limited inflammatory responses. Interestingly, both GRM affected the expression of genes related to antigen processing and presentation and in addition, GO activated pathways of neutrophil activation, degranulation and immunity in MDM. Overall, this study provides an extensive resource of potential toxicity mechanisms for future safety assessment of GRM in more advanced model systems to verify if the observed changes in gene expression in human macrophages could lead to long-term consequences on human health.

Release of graphene-related materials from epoxy-based composites: characterization, quantification and hazard assessment in vitro.

Due to their mechanical strength, thermal stability and electrical conductivity, graphene-related materials (GRMs) have been extensively explored for various applications. Moreover, GRMs have been studied and applied as fillers in polymer composite manufacturing to enhance the polymer performance. With the foreseen growth in GRM production, occupational and consumer exposure is inevitable, thus raising concerns for potential health risks. Therefore, this study aims (1) to characterize aerosol particles released after mechanical abrasion on GRM-reinforced epoxy composites, (2) to quantify the amounts of protruding and free-standing GRMs in the abraded particles and (3) to assess the potential effects of the pristine GRMs as well as the abraded particles on human macrophages differentiated from the THP-1 cell line in vitro. GRMs used in this study included graphene nanoplatelets (GNPs), graphene oxide (GO), and reduced graphene oxide (rGO). All types of pristine GRMs tested induced a dose-dependent increase in reactive oxygen species formation, but a decrease in cell viability was only detected for large GNPs at high concentrations (20 and 40 µg mL-1). The particle modes measured using a scanning mobility particle sizer (SMPS) were 300-400 nm and using an aerodynamic particle sizer (APS) were between 2-3 µm, indicating the release of respirable particles. A significant fraction (51% to 92%) of the GRMs embedded in the epoxy composites was released in the form of free-standing or protruding GRMs in the abraded particles. The abraded particles did not induce any acute cytotoxic effects.