The iron matters: Aged microplastics disrupted the iron homeostasis in the liver organoids

Plastic products undergo artificial and unintentional aging during daily use, causing the presence of aged microplastics (aMP). Humans are inevitably exposed to aMP. Liver is one of the critical target organs of MP through oral intake, however, limited research has focused on the hepatic toxicity of aMP compared to pristine MP (pMP). We utilized the human pluripotent stem cells-derived liver organoids (LOs) to compare the cytotoxicity of pristine polystyrene microplastics (pPS) (1 μm, carbonyl index 0.08) and aged polystyrene microplastics (aPS) (1 μm, carbonyl index 0.20) ranged from 20 to 200 ng/mL. Our findings indicate that aPS was more cytotoxic than pPS. We explored the disrupted iron homeostasis in terms of the [Fe2+] and [Fe3+] levels, iron storage and transport. A "vector-like effect" induced by aPS has been preliminarily suggested by the correlated change in total iron level and co-localization of PS and ferritin light chain (FTL) in the LOs following exposure to aPS and ferric ammonium citrate (FAC) individually and combinedly. In addition, we observed abnormal mitochondrial morphology, elevated lipid peroxidation, and declined GSH peroxidase activity, together with the declined expression of transferrin receptor (TFRC) and elevated expressions of SLC7A11, FTL. The gene handled iron transport and iron use were disrupted by aPS. Moreover, we employed FAC to introduce iron overload and Nacetylcysteine (NAC) to protect the lipid peroxidation. In aPS + FAC group, aggravated effects could be observed in aspects of [Fe2+] level, lipid peroxidation, and compromised expression levels of iron homeostasis-related markers, in contrast, in aPS + NAC group, most of changes recovered but the hepatocytoxicity remained. Specifically, a dimorphic change in elevated FTL and decreased ferritin heavy chain (FTH1) caused by 50 ng/mL aMP (57.33 ± 3.57 items/mL, equivalent to human intake level), indicated a specific response to low-dose aMP.