Toxicity enhancement of microplastics released from food containers through thermal aging: Absorbing more serum proteins thus activating the innate immune response via actin polymerization

This study examined the effects of hot high-fat simulants on the physicochemical properties of microplastics (MPs) from polypropylene (PP)-, low-density polyethylene (LDPE)-, and polylactic acid (PLA)-based single-use food container (SUFC) leachates and those of aging on their immunomodulatory effectors. Scenario studies have demonstrated that MPs were released from these three types of SUFCs. LDPE- and PLA-based SUFCs also released cellulose. Among the SUFCs, only the PP leachates particles exhibited a new absorption peak at 1725 cm^-1, which aging phenomenon may be attributed to the presence of unstable tertiary carbon atoms. Subsequently, we investigated the immunomodulatory effects of removing additive both PP and thermal-aged PP with polystyrene (PS) and carboxyl-modified PS (PS-COOH) polymer backbones as reference materials. The findings indicated that thermal-aged PP and PS-COOH induced comparable innate immune responses, with PS-COOH particles exhibiting a similar size to SUFC percolates. Consequently, PS and PS-COOH were selected as original and thermal-aged MPs, respectively, to evaluate the effects of aging on innate immunity. The results revealed thata protein corona formed on both particle types, with more protein adsorption observed on PS-COOH particles. The complex enhanced the phagocytosis of RAW264.7 macrophages and increased the expression of pro-inflammatory genes NOS2 and TNF-α through an actin polymerization cross-linking mechanism. In this study, we investigated how thermal-aged MPs affect innate immune responses using PS-COOH as a model system, emphasizing the importance of a comprehensive safety evaluations of MPs.