Unravelling immunomodulatory effects of bisphenol A substitutes on human macrophages, T and B lymphocytes using in vitro models

This study investigates the immunomodulatory effects of BPA substitutes using in silico and in vitro approaches. A comprehensive in silico screening with Endocrine Disruptome of BPA and its 25 substitutes revealed that BPG, BPS-MAE, BPS-MPE, BPPH and PF201 have the highest potential to bind nuclear receptors. Based on the in silico ranking, 12 compounds were selected and tested in vitro to investigate their effects on THP-1 derived macrophages, Jurkat T cells and LCLs. The metabolic activity tests showed that BPA has IC₅₀ values of 99-182 µM, while the values for BPG, BPPH and BPP are 5-10 times lower. Comparison of the IC₅₀ values with the logP of the tested compounds showed that the BPA analogues affect cell viability in proportion to their lipophilicity (R² = 0.9185). The results of the cytokine release assays showed that the BPA substitutes generally stimulated the proinflammatory response in THP-1 macrophages in a statistically significant manner when exposed to the environmentally relevant concentrations and suppressed it in the micromolar range in all cell lines tested. Further analysis using RamosBlue reporter cells indicated that BPAP, BPG, BPP and BPPH alter the NF-κB/AP-1 signalling pathway. Moreover, BPAP, BPG, BPP, BPPH, BPS-MAE, BPS-MPE, BTUM and PF201 were predicted in silico to be TLR4-MyD88 inhibitors. The overall results of this study identify BPG, BPP, BPPH, BPZ and TCBPA as BPA analogues with the highest immunomodulatory potential. In addition, the immunomodulatory potential of the BPA alternatives Pergafast201 and BTUM was confirmed in vitro for the first time. This study emphasises the need for cautious evaluation of BPA substitutes due to their potential immunomodulatory effect, which could significantly affect public health.