Nanoplastics (NPs) are one of the most abundant environment-threatening nanomaterials on the market. The objective of this study was to determine in vitro if functionalized NPs are cytotoxic by themselves or increase the toxicity of metals. For that, we used 50 nm polystyrene nanoparticles with distinct surface functionalization (pristine, PS-Plain; carboxylic, PS-COOH; and amino PS-NH₂) alone or combined with the metals arsenic (As) and methylmercury (MeHg), which possess an environmental risk to marine life. As test model, we chose a brain-derived cell line (SaB-1) from gilthead seabream (Sparus aurata), one of the most commercial fish species in the Mediterranean. First, only the PS-NH₂ NPs were toxic to SaB-1 cells. NPs seem to be internalized into the cells but they showed little alteration in the transcription of genes related to oxidative stress (nrf2, cat, gr, gsta), cellular protection against metals (mta) or apoptosis (bcl2, bax). However, NPs, mainly PS-COOH and PS-NH₂, significantly increased the toxicity of both metals. Since the coexistence of NPs and other pollutants in the aquatic environment is inevitable, our results reveal that the combined effect of NPs with the rest of pollutants deserves more attention.