Effects of polystyrene micro/nanoplastics on liver cells based on particle size, surface functionalization, concentration and exposure period

Micro/nanoplastics (MP/NP) contaminate our food and drinking water but their impact on human health has not been well-documented. The liver is one of the first organs that ingested MP/NP encounter and it has a major role in the clearance of xenobiotics. Therefore, the effects of polystyrene MP/NP on liver HepG2 cells were studied. Cellular responses to particles of various sizes (50-5000 nm) and surface functionalization (aminated, carboxylated or non-functionalized) were determined at different concentrations (0.1-100 μg/mL) and exposure periods (1-24 h). Smaller sized particles were internalized by HepG2 cells more avidly than larger particles regardless of functionalization; the highest uptake being for 50 and 100 nm aminated particles at lower concentrations. Confocal microscopy images of cells corroborated quantitative uptake results. Aminated particles were more toxic to the cells than carboxylated or non-functionalized particles. Among aminated particles smaller particles (50 and 100 nm) were more detrimental to cell viability compared to larger particles (1000 or 5000 nm) with toxicity increasing with concentration. Treatment with the particles for 4 h increased intracellular concentrations of Caspase-3 by 1.5-2.8 fold, but 24 h exposure to the particles attenuated this increase in Caspase-3 concentrations. A slight trend of higher Caspase-3 concentration in cells treated with larger particles (500-5000 nm) compared to smaller particles (50-200 nm) was observed, indicating that larger particles are more likely to direct cells toward apoptotic cell death upon 4 h exposure. Exposure of cells to large PS particles (500-5000 nm) upregulated interleukin-8 and the effect was enhanced at 24 h. Overall, the study demonstrated that smaller aminated particles were most toxic to hepatocytes, but larger particles induced apoptotic cell death or an inflammatory response depending on the length of exposure.