Perfluorodecanoic acid (PFDA), a long-chain perfluoroalkyl substance (PFAS) known for its environmental persistence and toxicity, and dimethyl sulfoxide (DMSO), a commonly used solvent in biological research, both have notable impacts on cellular function. However, the molecular pathways underlying their effects, particularly in non-mammalian cells such as RTgill-W1 derived from rainbow trout gill tissues, remain insufficiently understood. This study investigated the effects of DMSO and PFDA on RTgill-W1 cells using high-throughput metabolomics and lipidomics, alongside cell viability and oxidative stress assays.
DMSO exposure (0.1–10%) resulted in dose-dependent declines in cell viability starting at 0.5% and significant increases in reactive oxygen species (ROS) production at concentrations ≥4%. Metabolomic analysis revealed disruptions in 41 metabolic pathways across five functional groups—amino acid, carbohydrate, lipid, nucleotide, and vitamin metabolism—with pronounced effects at higher concentrations. Even low DMSO concentrations (≤0.5%) elicited widespread metabolic changes, highlighting the need for including proper solvent controls in in vitro studies.
PFDA exposure (1, 10, 20, 40, 60, 80, 160 and 320 mg/L) exhibited dose-dependent cytotoxicity, with an EC₅₀ of 51.9 ± 1.7 mg/L. Significant increase in ROS production was observed at 40 mg/L and higher, indicating elevated oxidative stress. Metabolomic profiling revealed disruptions of PFDA in 190 metabolites across key metabolic pathways, while lipidomic analysis identified 102 significantly altered lipids. These changes implicated disruptions in glycerolipid and glycerophospholipid metabolism, sphingolipid pathways, and fatty acid biosynthesis, suggesting compromised membrane integrity, energy production, and signalling.
This study highlights the broad biochemical impacts of DMSO and PFDA on fish cell lines and underscores their capacity to interfere with critical cellular processes. The findings reinforce the utility of integrated omics approaches in ecotoxicological research and emphasize caution when using DMSO as a solvent. Future studies should validate these findings through short-term in vivo assays to improve ecological relevance and reliability.