Honza Tatiana, El Yamani Naouale, Dusinska Maria, Rundén-Pran Elise, Marcon Francesca
The Climate and Environmental Research Institute (NILU) Norway.
Italian Institute of Health (ISS) Italy.
EFSA J. 2024 Dec 20;22(Suppl 1):e221116. doi: 10.2903/j.efsa.2024.e221116. eCollection 2024 Dec.
The safety of bisphenol A (BPA) due to its adverse effects on the immune system has led to an increasing concern and a significant regulatory shift. The European Food Safety Authority (EFSA) proposed a reduction in the tolerable daily intake (TDI) of BPA in food in their 2023 scientific opinion, highlighting the need for stricter regulations compared to their previous assessment in 2015. This regulatory action has spurred the production of BPA alternatives, raising concerns about their safety due to insufficient toxicological data. Addressing this gap is crucial for ensuring human and environmental health. In this project, multiple genotoxicity endpoints were applied for testing of two regulatory relevant BPA alternatives, bisphenol E (BPE) and bisphenol P (BPP), in different human models: 2D HepG2 liver cells, 3D liver spheroids and primary human peripheral blood lymphocytes. DNA strand breaks and oxidised base lesions were evaluated by the enzyme-modified version of the comet assay, while clastogenicity and aneugenicity were analysed by the in vitro micronucleus assay (OECD TG 487, 2016), together with cytotoxicity. Development of new advanced models (NAMs), as 3D spheroids, are essential for next-generation risk assessment (NGRA) in line with the 3R's to replace, reduce or refine animal experiments. In this aspect, validation and standardisation of NAMs are needed to reach regulatory readiness level and development of OECD Test Guidelines. Therefore, a standardisation and pre-validation of the advanced 3D liver spheroid model was performed by using multiple genotoxicity endpoints and by comparing the obtained results with standard genotoxicity models.
双酚A（BPA）因其对免疫系统的不良影响，安全性引发了越来越多的关注，监管也发生了重大转变。欧洲食品安全局（EFSA）在其2023年的科学意见中提议降低食品中BPA的每日可耐受摄入量（TDI），强调与2015年的先前评估相比，需要更严格的监管。这一监管行动刺激了BPA替代品的生产，但由于毒理学数据不足，人们对其安全性表示担忧。填补这一空白对于确保人类和环境健康至关重要。在本项目中，在不同的人类模型中应用了多个遗传毒性终点来测试两种与监管相关的BPA替代品，双酚E（BPE）和双酚P（BPP）：二维HepG2肝细胞、三维肝脏球体和原代人外周血淋巴细胞。通过彗星试验的酶修饰版本评估DNA链断裂和氧化碱基损伤，同时通过体外微核试验（经合组织TG 487，2016）分析致断裂性和非整倍体性，并分析细胞毒性。作为三维球体的新型先进模型（NAMs）的开发对于符合3R原则（替代、减少或优化动物实验）的下一代风险评估（NGRA）至关重要。在这方面，需要对NAMs进行验证和标准化以达到监管准备水平并制定经合组织测试指南。因此，通过使用多个遗传毒性终点并将获得的结果与标准遗传毒性模型进行比较，对先进的三维肝脏球体模型进行了标准化和预验证。
