Mixture toxicity effects and uptake of titanium dioxide (TiO₂) nanoparticles and 3,3″,4,4″-tetrachlorobiphenyl (PCB77) in juvenile brown trout following co-exposure via the diet

Titanium dioxide nanoparticles (n-TiO₂) are among the man-made nanomaterials that are predicted to be found at high concentrations in the aquatic environment. There, they likely co-exist with other chemical pollutants. Thus, n-TiO₂ and other chemical pollutants can be taken up together or accumulate independently from each other in prey organisms of fish. This can lead to dietary exposure of fish to n-TiO₂-chemical pollutant mixtures. In this study, we examine if simultaneous dietary exposure to n-TiO₂ and 3,3’,4,4’-Tetrachlorobiphenyl (PCB77) –used as a model compound for persistent organic pollutants with dioxin-like properties– can influence the uptake and toxicological response elicited by the respective other substance. Juvenile brown trout (Salmo trutta) were fed custom-made food pellets containing n-TiO₂, PCB77 or n-TiO₂+PCB77 mixtures for 15 days. Ti and PCB77 concentrations in the liver were measured by ICP-MS and GC–MS, respectively. Besides, n-TiO₂ uptake was assessed using TEM. Combination effects on endpoints specific for PCB77 (i.e., cytochrome P450 1A (CYP1A) induction) and endpoints shared by both PCB77 and n-TiO₂ (i.e., oxidative stress-related parameters) were measured in intestine and liver using RT-qPCR and enzyme activity assays. The results show that genes encoding for proteins/enzymes essential for tight junction function (zo-1) and ROS elimination (sod-1) were significantly upregulated in the intestine of fish exposed to n-TiO₂ and PCB77 mixtures, but not in the single-substance treatments. Besides, n-TiO₂ had a potentiating effect on PCB77-induced CYP1A and glutathione reductase (GR) expression/enzyme activity in the liver. This study shows that simultaneous dietary exposure to nanomaterials and traditional environmental pollutants might result in effects that are larger than observed for the substances alone, but that understanding the mechanistic basis of such effects remains challenging.