Size-dependent neurotoxicity of micro- and nanoplastics in flowing condition based on an in vitro microfluidic study

With the widespread presence of plastic wastes, knowledge about the potential environmental risks and bioavailability of micro- or nanoplastics fragmented from large analogs is of utmost importance. As the particle size matters in mediating endocytic mechanism and particle internalization, we first studied the effects of polystyrene microparticles (PS-MPs, 1 μm) and polystyrene nanoparticles (PS-NPs, 100 nm) of two different sizes at varying concentrations of 5, 25 and 75 μg/mL on the mouse hippocampal neuronal HT22 cells. The in vitro study showed efficient cellular uptake of PS-MPs and PS-NPs of both sizes. The adverse effects of cellular metabolic activity as reflective of excess Reactive Oxygen Species (ROS) and cell cycle S phase arresting were observed especially at the greater concentration of smaller-sized PS particles, consequently leading to mild cytotoxicity. We further evaluated the dynamic particle-cell interaction with a continuous supply of PS particles using a microfluidic device. By recapitulating the in vivo mechanical microenvironments while allowing homogeneous distribution of PS particles, the dynamic exposure to PS particles of both sizes under flowing conditions resulted in much lesser viability of neural cells than the traditional static exposure. As the flowing dynamics may avoid the gravitational settling of particles and allow more efficient cellular uptake, the size distribution, together with the exposure configurations, contributed significantly to the determination of the PS particle cytotoxicity. The on-chip investigation and a better understanding of particle translocation mechanisms would offer very much to the risk assessment of PS particles on human health.