Titanium dioxide nanoparticles (TiO₂ NPs) as food additives were widely found in various foodrelated products, especially in high-sugar foods. The daily intake of TiO₂ NPs in the diet may therefore expose the small intestine to TiO₂ NPs and affect its physiological functions, including the absorption of nutrients. It is speculated that TiO₂ may cause serious health hazards by increasing sugar uptake. To explore this possibility, transport of glucose from small intestine was studied using an everted gut sac model prepared from small intestine of young healthy male SD rats. The translocation of TiO₂ NPs and the morphological changes of small intestine were also observed after exposure of intestinal lumen to TiO₂ NPs for 2 h. The results showed that TiO₂ NPs can enter into enterocyte but hardly cross the intestinal epithelium. No change on microstructure of gut epithelia and expression of glucose transporter was found, and there is no obvious impact on intestinal absorption and metabolism of glucose. These results suggest that short-term exposure to TiO₂ NPs has little influence on intestinal absorption of glucose. More attention should be paid to the chronic effect of dietary consumption of TiO₂ NPs on nutrient absorption.