Cellular toxicity assessment of common plasticizers: Implications for musculoskeletal and cardiovascular health

The cardiovascular toxicity of plasticizers remains insufficiently characterized, despite increasing evidence linking environmental chemical exposures to vascular dysfunction and cardiometabolic disease. Regulatory restrictions on legacy plasticizers have accelerated the use of structurally diverse alternative compounds, many introduced with limited human-relevant cardiovascular hazard data. This study evaluated the cardiovascular cellular stress effects of legacy plasticizers, di(2-ethylhexyl) phthalate (DEHP) and bisphenol A (BPA), and alternatives, including di(2-ethylhexyl) adipate (DEHA), dioctyl adipate (DOA), and trioctyl trimellitate (TOTM), using human in vitro vascular models. Human endothelial (HMEC-1), skeletal muscle (RMS-13), and aortic vascular smooth muscle (T/G HA-VSMC) cells were assessed for cytotoxicity, proliferation/migration, reactive oxygen species (ROS), mitochondrial membrane potential (ΔΨm), and intracellular calcium dynamics. BPA was the only compound to induce overt cytotoxicity across cell types. However, several plasticizers produced significant sub-cytotoxic alterations in oxidative stress regulation, mitochondrial function, and calcium homeostasis. Alternative plasticizers reduced intracellular ROS and calcium levels, while ΔΨm responses were cell-type dependent, revealing vascular stress signatures not detected by traditional cytotoxicity screening. These findings demonstrate that both legacy and alternative plasticizers can disrupt critical cardiovascular pathways and underscore the utility of NAM-based vascular models for chemical safety assessment and regulatory decision-making.