Titanium dioxide nanoparticles perturb the blood-testis barrier via disruption of actin-based cell adhesive function

As one of the most commonly used nanoparticles, titanium dioxide nanoparticles (TiO₂-NPs) are widely used as coating reagents in cosmetics, medicine and other industries. The increasing risk of exposure to TiO₂-NPs raises concerns about their safety. In this study, we investigated the mechanism by which TiO₂-NPs cross the blood-testis barrier (BTB). TM-4 cells were selected as an in vitro Sertoli cell model of BTB. Cell viability, cell morphological changes, apoptosis, oxidative damage, and the expression levels of actin regulatory and tight junction (TJ) proteins were assessed in TM-4 cells treated with 3-nm and 24-nm TiO₂-NPs. Cells treated with 3-nm TiO₂-NPs exhibited increased cytotoxicity and decreased Annexin II expression, whereas cells treated with 24-nm TiO₂-NPs exhibited increased Arp 3 and c-Src expression. Both TiO₂-NPs induced significant oxidative stress, decreased the expression of TJ proteins (occludin, ZO-1 and claudin 5), damaged the TJ structure, and exhibited enlarged gaps between TM-4 cells. Our results indicated that both TiO₂-NPs crossed the BTB by disrupting actin-based adhesive junctions of TM-4 cells; however, apoptosis was not observed. Our results provide new insights into how TiO₂-NPs cross the BTB.