Hexavalent chromium amplifies the developmental toxicity of graphene oxide during zebrafish embryogenesis

Combined toxicity is a critical issue in risk assessment of contaminants. However, very little is known about the joint effects of graphene oxide (GO, a crucial 2-dimensional carbon material) and hexavalent chromium (Cr⁶⁺, a widespread heavy metal), particularly with respect to the critical period of embryogenesis. In this study, the combined toxicity of GO and Cr⁶⁺ was evaluated through embryo-larval toxicity test in Danio rerio (zebrafish). Results indicated that the co-exposure of Cr⁶⁺ (1 mg/L) and GO (0.01 mg/L) inhibited hatching and spontaneous movement of embryos, but no significant changes were found in the single Cr⁶⁺ or GO group. Compared with the single GO or Cr⁶⁺ exposure, their co-exposure (GO+Cr⁶⁺) significantly enhanced the teratogenicity in a concentration-dependent pattern, and the spinal curvature was observed as the main deformity. GO+Cr⁶⁺ changed the protein secondary structures of embryos result of the generation of ROS and oxidative stress. The degradations of vertical myosepta and cartilages were observed in co-exposure group, suggesting that GO+Cr⁶⁺ disrupted the development of musculoskeletal system. The genes col11a1a, col2a1a and postnb were down-regulated but the genes acta1b and mmp9 were up-regulated by GO+Cr⁶⁺. The interactions between Cr⁶⁺ and GO demonstrated that the morphology, structure, and surface properties of GO were modified by Cr⁶⁺. The enhanced defects and O-containing groups of GO could trap more β-sheets, induced oxidative stress, disturbed the development of skeletal muscles and cartilages in zebrafish. These data suggested that GO+Cr⁶⁺ enhanced their joint toxicity due to the variation of nanoparticle properties. This finding is important for assessing the ecological risk of graphene family nanomaterials in the natural environment.