TiO2 nanosheets promote the transformation of vascular smooth muscle cells into foam cells in vitro and in vivo through the up-regulation of nuclear factor kappa B subunit 2

Titanium dioxide (TiO2) nanomaterials have been shown to promote atherosclerosis through endothelial dysfunction. This study investigated the toxicity of TiO2 nanosheets (NSs) to vascular smooth muscle cells (VSMCs), one of the pivotal cells involved in all stages of atherosclerosis. Only a high concentration of TiO2 NSs (128 μg/mL) modestly induced cytotoxicity by decreasing thiols. RNA-sequencing data revealed that 64 μg/mL TiO2 NSs significantly down-regulated 94 genes and up-regulated 174 genes, respectively. Gene ontology (GO) terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways related to SMC function and lipid metabolism were altered. TiO2 NSs increased nuclear factor kappa B subunit 2 (NFKB2), which led to a decrease in VSMC marker actin alpha 2, smooth muscle (ACTA2). On the other hand, macrophage marker CD36 and fatty acid synthase (FASN) proteins were increased. Additionally, TiO2 NSs induced inflammatory cytokines and lipid accumulation, and these effects were curtailed by NFKB inhibitor - triptolide. Furthermore, repeated TiO2 NS injection (5 mg/kg BW, once a day for 5 continuous days) into ICR mice led to increased NFKB2, CD36 and FASN, with a decreased ACTA2. Our results suggested that TiO2 NSs promoted the transformation of VSMCs into foam cells through the up-regulation of NFKB2.