Cellular internalization pathways of environmentally exposed microplastic particles: Phagocytosis or macropinocytosis?

Microplastic particles (MP) ubiquitously occur in all environmental compartments where they interact with biomolecules, forming an eco-corona on their surfaces. The eco-corona affects the surface properties of MP and consequently how they interact with cells. Proteins, an integral component within the eco-corona, may serve as a ligand driving the interaction of MP with membrane receptors. To date, it is not known, whether eco-coronae originating from different environmental media differ in their proteinaceous compositions and whether these particles interact differently with cells. We show that the protein composition of the eco-coronae formed in freshwater (FW) and salt water (SW) are distinct from each other. We did not observe different adhesion strengths between MP coated with different eco-coronae and cells. However, the internalization efficiency and the underlying internalization mechanisms significantly differed between FW- and SW eco-coronae. By inhibiting actin-driven and receptor-mediated internalization processes using Cytochalasin-D, Amiloride, and Amantadine, we show that FW microplastic particles predominantly become internalized via phagocytosis, while macropinocytosis is more important for SW microplastic particles. Overall, our findings show that the origin of eco-coronae coatings are important factors for the cellular internalization of microplastic particles. This highlights the relevance of eco-coronae for adverse effects of environmentally relevant microplastic particles on cells and organisms.