Tri-o-tolyl phosphate impedes implantation: Malfunction of mitochondria and disruption of calcium homeostasis through MAPK and AKT signaling cascades.

Tri-o-tolyl phosphate (TOTP), a flame retardant containing aryl compounds, is widely used in human living environments owing to its several applications. However, Due to the overuse of TOTP, its residue has been identified in various environments and non-targeted organisms, including humans. Although extensive research is being conducted to address the toxicity of this substance, its potential reproductive toxicity in females has not been sufficiently studied. In this study, human HTR-8/SVneo and JEG-3 trophoblasts were used to investigate the effects of TOTP on implantation. Results showed that TOTP decreased cell viability and inhibited cell proliferation by triggering cell cycle arrest. It also induced apoptosis and mitochondrial dysfunction, disrupted calcium homeostasis, increased the influx of calcium ions into the mitochondria, and disturbed cell aggregation and migration. Moreover, the MAPK and AKT cell signaling pathways were altered, and crosstalk between these pathways were distinguished. Thus, inhibitors of the MAPK and AKT pathways exhibited potential for managing the toxicity of TOTP. Overall, this study demonstrated the reproductive toxicity of TOTP in human females and elucidated the underlying mechanisms. Our results highlighted the potential risks associated with TOTP.