BPA, BPAF and TMBPF Alter Adipogenesis and Fat Accumulation in Human Mesenchymal Stem Cells, with Implications for Obesity

Abstract

Bisphenol A (BPA) is an endocrine-disrupting chemical used in the production of plastics, and is linked to developmental, reproductive, and metabolic disorders including obesity. Manufacturers have begun using ‘BPA-free’ alternatives instead of BPA in many consumer products. However, these alternatives have had much less testing and oversight, yet they are already being mass-produced and used across industries from plastics to food-contact coatings. Here, we used human female adipose-derived stem cells (hASCs), a type of adult mesenchymal stem cell, to compare the effects of BPA and BPA alternatives on adipogenesis or fat cell development in vitro. We focused on two commonly used BPA replacements, bisphenol AF (BPAF) and tetramethyl bisphenol F (TMBPF; monomer of the new valPure V70 food-contact coating). Human ASCs were differentiated into adipocytes using chemically defined media in the presence of control differentiation media with and without 17β-estradiol (E2; 10 μM), or with increasing doses of BPA (0, 0.1 and 1 μM), BPAF (0, 0.1, 1 and 10 nM), or TMBPF (0, 0.01 and 0.1 μM). After differentiation, the cells were stained and imaged to visualize and quantify the accumulation of lipid vacuoles and number of developing fat cells. Treated cells were also examined for cell viability and apoptosis (programmed cell death) using the respective cellular assays. Similar to E2, BPA at 0.1 μM and BPAF at 0.1 nM, significantly increased adipogenesis and lipid production by 20% compared to control differentiated cells (based on total lipid vacuole number to cell number ratios), whereas higher levels of BPA and BPAF significantly decreased adipogenesis (P < 0.005). All tested doses of TMBPF significantly reduced adipogenesis and lipid production by 30–40%, likely at least partially through toxic effects on stem cells, as viable cell numbers decreased and apoptosis levels increased throughout differentiation. These findings indicate that low, environmentally-relevant doses of BPA, BPAF, and TMBPF have significant effects on fat cell development and lipid accumulation, with TMBPF having non-estrogenic, anti-adipogenic effects. These and other recent results may provide a potential cellular mechanism between exposure to bisphenols and human obesity, and underscore the likely impact of these chemicals on fat development in vivo.

Teratogenicity and toxicity of the new BPA alternative TMBPF, and BPA, BPS, and BPAF in chick embryonic development

“…Hence, TMBPF is being marketed as a 'low toxicity' BPA replacement monomer. However, we and others have found that TMBPF does have toxic effects on human and rat stem cells, and other human cell lines, and at very low concentrations, calling into question the safety of this new chemical ( Cohen et al, 2021 , Harnett et al, 2021a , Harnett et al, 2021b , Szafran et al, 2017 ).…”

“…Similar to BPA, many of them are likely EDCs that interfere with hormone receptors and action. Numerous studies reveal the toxic potential of BPA analogs, showing various cellular, developmental, and reproductive disruptions across species ( Arancio et al, 2018 , Arancio et al, 2019 , Cohen et al, 2021 , Harnett et al, 2021a , Harnett et al, 2021b , Eladak et al, 2015 , Ge et al, 2014 , Huang et al, 2016 , Moreman et al, 2017 , Wang et al, 2017 , Wu et al, 2017 ). Importantly, studies in various animal models and cell systems report that some analogs such as BPAF, are even more toxic, potent, and estrogenic than the parent compound BPA ( Arancio et al, 2018 , Cohen et al, 2021 , Kojima et al, 2019 , Liang et al, 2017 , Michałowicz et al, 2015 , Moreman et al, 2017 ).…”

Using inhibition of the adipogenesis of adipose-derived stem cells in vitro for toxicity prediction

“…In summary, researchers can use the protocol described here to evaluate cytotoxicity during cell differentiation processes in vitro and for other applications, such as the study of the basic biology of stem cells or the screening of test substances that may interfere with adipogenesis [29] , [30] , [31] .…”

Effects of Bisphenol A and Its Alternatives, Bisphenol F and Tetramethyl Bisphenol F on Osteoclast Differentiation

“…It was shown to not produce any estrogenic effects in both in vitro and in vivo [33,51]. According to a recent study, TMBPF has effects on adipogenesis and fat accumulation, having nonestrogenic, antiadipogenic effects [52]. Moreover, TMBPF revealed antiestro-genic and antiandrogenic activity in several cancer cell lines [33,53].…”