In an effort to mitigate the adverse health effects of the legacy compound bisphenol A (BPA), the use of alternatives has been increasing. Unfortunately, there is little information on the toxicity of some of these chemicals. Since many are structural analogues of BPA, we have compared the effects of BPA and five of its alternatives, BPAF, BPF, BPS, BPM, and BPTMC, using a high-content imaging system. Phenotypic parameter data were then analysed using both benchmark concentration (BMC) analyses and administered equivalent dose (AED) approaches. Assessment of cell lines associated with the female and male reproductive systems showed evidence of phenotypic parameter changes. When assessing C18-4 (spermatogonial stem cells), MA-10 Leydig cells, and KGN granulosa cells, BPA was one of the least cytotoxic compounds tested. BPAF was the most cytotoxic to MA-10 cells, followed by C18-4 and KGN cells. Interestingly, BPF and BPS were cytotoxic only to MA-10 cells. BPM and BPTMC were both highly cytotoxic in all three cell lines studied. High-content imaging analyses revealed chemical and cell-line specific effects on phenotypic parameters such as lipid droplets and oxidative stress. In vitro to in vivo extrapolation (IVIVE) analyses revealed that these effects occurred at AEDs ranging from 0.2 to 16.5 mg/kg body weight/day. In summary, BPA replacements altered unique phenotypic parameters in addition to cytotoxic effects. These data suggest that this approach can aid in the screening and selection of responsible replacements for downstream testing and analysis.

Supported by CIHR and McGill University.