Comparative evaluation of bisphenol A, bisphenol F and tetramethyl bisphenol F on adipogenesis, oxidative stress and inflammation in 3T3-L1 adipocytes

Bisphenol A (BPA) has long been recognized as an endocrine-disrupting chemical (EDC) with obesogenic properties, prompting widespread industrial substitution with structural analogues, among which bisphenol F (BPF) and tetramethyl bisphenol F (TMBPF) have gained particular commercial prominence. However, the metabolic safety of these alternatives remains inadequately characterized. The present study systematically compared the adipogenic, pro-oxidative, and pro-inflammatory effects of BPA, BPF, and TMBPF in differentiated 3T3-L1 murine preadipocytes and employed in silico molecular docking to elucidate their interactions with key nuclear receptors. Treatment with all three compounds at concentrations of 0.01–1 µM during adipocyte differentiation induced dose-dependent increases in intracellular lipid accumulation and lipid droplet size, with TMBPF eliciting the most pronounced effect (approximately 300% of MDI control at 1 µM). Consistently, mRNA and protein expression of the master adipogenic transcription factors PPARγ and C/EBPα, together with lipid metabolism markers including SREBP-1c, FABP4, FAS, and LPL, were significantly upregulated by all three compounds in a concentration-dependent manner. TMBPF also exhibited the strongest binding affinity for the PPARγ ligand-binding domain (docking score: −8.4 kcal/mol) and the glucocorticoid receptor (GR; −8.3 kcal/mol), consistent with its potent in vitro adipogenic activity. Adipokine profiling revealed increased expression of adiponectin, leptin, and resistin, with TMBPF treatment resulting in the most prominent upregulation of AdipoR1. Furthermore, TMBPF selectively suppressed antioxidant enzyme expression ( Cat , Sod2 , and Gpx1 ) and all three bisphenols markedly elevated secretion of pro-inflammatory cytokines TNF-α, IL-6, and MCP-1. Collectively, these findings indicate that TMBPF, despite its classification as a “safer” BPA alternative, exhibits greater adipogenic and oxidative disrupting potency compared to BPA and BPF, raising concerns that warrant further in vivo and epidemiological evaluation of TMBPF, particularly given its increasing use as a food-contact coating material.