In order to evaluate the penetration of the nanoparticles, eight adult male Wistar rats (3 months old) were used for the in vivo experiments. The protocol was approved by the local University Committee for animal testing and is in accordance with the Canadian Council on Animal Care (CICUAL-RD-2021–892-E-UNC-DEC#FCQ).

A study published in the Journal of Agricultural and Food Chemistry in 2019 sought to examine the effects of titanium dioxide on intestinal inflammation. Researchers did this by feeding rats titanium dioxide nanoparticles and found that, after the course of two to three months, the animals had lower body weights and induced intestinal inflammation. The researchers also found the nanoparticles altered gut microbiota composition and aggravated chronic colitis. The rats also experienced reduced populations of CD4+T cells (which are cells that help organize immune responses by prompting other immune cells to fight infection), regulatory T cells, and white blood cells in mesenteric lymph nodes. The researchers wrote: “Dietary TiO2 nanoparticles could interfere with the balance of the immune system and dynamic of gut microbiome, which may result in low-grade intestinal inflammation and aggravated immunological response to external stimulus, thus introducing potential health risk.”