No acute effects of nano-sized TiO₂ were observed in Danio rerio (zebrafish) embryos. Exposure of rainbow trout to TiO₂ NPs triggered lipid peroxidation, influence on the respiratory tract, disturbance in the metabolism of Cu and Zn, induction of intestinal erosion  and accumulation in kidney tissue. Linhua et al. exposed juvenile carp to 100 and 200 mg/ml of particles and TiO₂ observed no mortality. However, the fish suffered from oxidative stress and pathological changes in gill and liver. In the infaunal species Arenicola marina, exposure to TiO₂ NPs in sediment caused sub-lethal effects including decrease in casting rate and increase in cellular and DNA damage. Aggregated particles were visible in the lumen of the gut, but no uptake through the gut or the skin was observed.

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In a small study published in the European Journal of Nutrition in 2020, researchers examined the effects of several food additives, including titanium dioxide, along with artificial sweeteners and cleaning products by testing the fecal samples of 13 people. Titanium dioxide was among the samples that “induced significant shifts in microbiome community structure.”  The growth of the bacterium species belonging to C. leptum, which has been shown to decrease in patients with inflammatory bowel disease, “significantly decreased in the presence of … titanium dioxide” among other additives and sweeteners tested.

A few non-dietary studies have reported adverse effects in the gastrointestinal tract of laboratory animals given food-grade TiO₂. However, these same effects were not seen when the same or higher doses of food-grade TiO₂ were administered in the animals' diet. Dietary studies best reflect how humans are exposed to TiO₂ from food. Thus, the Food Directorate placed the most emphasis on the results of these studies in the state of the science report.

There are numerous manufacturers of titanium dioxide; the largest include Delaware-based Chemours (a spin-off of DuPont Chemical), Texas-based Kronos, and China-based Lomon Billions Group, all of which manufacture pigments for use in products like paints, coatings, and plastics. UK-based Venator is a major supplier of titanium dioxide used in food and cosmetics, along with paints, paper, plastic, and more. As a pigment, it is called Pigment White 6 (PW6), titanium white, or CI 77891. As a food additive, it is known as E171.

Based on this opinion, the European Commission and the Member States agreed to remove all uses of titanium dioxide as an additive in food. In January 2022, a Regulation withdrawing the authorisation to use titanium dioxide as a food additive in food products was adopted i.e. Commission Regulation (EU) 2022/63.