Combined effect of polystyrene microplastics and bisphenol A on the human embryonic stem cells-derived liver organoids: The hepatotoxicity and lipid accumulation.

Human are exposed to microplastics (MP) via inhalation or ingestion daily and inevitably. The liver is an important target organ of MP. Bisphenol A (BPA) is one of commonly used plasticizers. It is added in plastics, but also generally detected in the biological samples of human beings. However, the combined toxic effect of MP and BPA on human liver is unclear. In this study, a novel 3D in vitro model, the liver organoid (LO) derived from human-pluripotent stem cells, has been utilized to explore the 1 μm polystyrene (PS)-induced hepatotoxicity with BPA individually and jointly. Conclusively, all the changes in the cytotoxicity, cellular and molecular makers regarding the energy supplement, hepatic injury, oxidative stress, inflammatory response, disruption in the lipid accumulation, as well as epigenetics regulation induced by BPA or PS on the LOs individually were comparable to previous study. The BPA levels in the culture medium were declined by the added PS. The combined adverse effect of PS and BPA on the LOs was identified to be synergistic upon deteriorated hepatotoxicity and interfered the gene panels related to multiple processes of lipid metabolism, together with the proteins of HNF4A, CD36, ACC1, CPT1A, CYP2E1, ERα and ERβ. Specifically, PS didn't change the ERα or ERβ individually, but when the LOs were co-exposed to PS and BPA, the ERα further elevated significantly and synergistically. Our findings highlight the metabolic-related health risk due to co-exposure to MP and BPA, even at low-doses equivalent to human internal exposure level. Based on these findings, the potential adverse outcome pathway related to PS and BPA singly and jointly were proposed, predicting two possible outcomes to be hepatic steatosis. Moreover, the ERα and HNF4A were proposed to be potential candidate markers to investigate the "vector-like effect" of PS in the present of BPA.