An integrated in vitro approach to identifying chemically induced oxidative stress and toxicity in mitochondria

Growing concerns exist about increasing chemical usage and the potential health risks. Developing an efficient strategy to evaluate or predict the toxicity of chemicals is necessary. The mitochondria are essential organelles for cell maintenance and survival but also serve as one of the main targets of toxic chemicals. Mitochondria play an important role in the pathology of respiratory disease, and many environmental chemicals may induce impairment of the respiratory system through mitochondrial damage. This study aimed to develop integrated in vitro approaches to identify chemicals that could induce adverse health effects by increasing mitochondria-mediated oxidative stress using the H441 cells, which have a club-cell-like phenotype. Twenty-six environmental toxicants (biocides, phthalates, bisphenols, and particles) were tested, and each parameter was compared with eleven reference compounds. The inhibitory concentrations (IC20 and IC50) and benchmark doses (BMD) of the tested compounds were estimated from three in vitro assays, and the toxic concentration was determined. At the lowest IC20, the effects of compounds on mitochondrial reactive oxygen species (ROS) production and mitochondrial membrane potential (MMP) were compared. Principal component analysis and k-mean clustering were performed to cluster the chemicals that had comparable effects on the cells. Chemicals that induce mitochondrial damage at different concentrations were used for an in-depth high-tier assessment and classification as electron transport system (ETS) uncoupling or inhibiting agents. Additionally, using in vitro to in vivo extrapolation (IVIVE) tools, equivalent administration doses and maximum plasma concentrations of tested compounds in human were estimated. This study suggests an in vitro approach to identifying mitochondrial damage by integrating several in vitro toxicity tests and calculation modeling.