Size- and Oxidation-Dependent Toxicity of Graphene Oxide Nanomaterials in Embryonic Zebrafish

Graphene oxides (GOs) are a popular graphene alternative. The goal of this study was to compare the biocompatibility of a diversity of well-characterized GOs. Our previous work advanced developmental zebrafish as a model to interrogate the interactions and biological responses following exposures to engineered nanomaterials (ENMs). Here, we investigated GO 250 nm × 250 nm (sGO), 400 nm × 400 nm (mGO), and 1 μm × 1 μm (lGO), partially reduced GO (prGO) 400 nm × 400 nm, and reduced GO (rGO) 400 nm × 400 nm and 2 μm × 2 μm, which first underwent extensive characterization under the support of the Nanomaterials Health Implications Research (NHIR) Consortium. GOs were stabilized in water (GOs), while prGO and rGOs were dispersed in sodium cholate. Zebrafish were statically exposed to up to 50 μg/mL of each material from 6 h post-fertilization (hpf) until 120 hpf. Toxicity was dependent on GO properties. mGO was the most toxic material; its effects manifested in the yolk syncytial layer (YSL). Additionally, sodium cholate stabilization significantly increased GO toxicity. The observed effects were size- and oxidation-state-dependent, revealing the importance of identifying the structure-specific toxicity of GOs.