Role of heteroaggregation and internalization in the toxicity of differently sized and charged plastic nanoparticles to freshwater microalgae

The toxic effect of waterborne nanoplastics is a manifestation of bio-nano interfacial interactions. Although nanoplastics with different physicochemical characteristics are known to exhibit distinct toxicities, it remains poorly understood how the properties of nanoplastics affect the bio-nano interface interactions. Here, polystyrene nanoparticles (PSNPs) varying in size (50, 300, and 500 nm) and surface charge (negative and positive charge) were employed to explore the interplay between PSNPs and algal cells (Chlamydomonas reinhardtii), with special focus on the heteroaggregation of PSNPs and microalgae, PSNPs cellular internalization, and cellular physiological responses. Results showed that large-sized PSNPs (300 and 500 nm) caused apparent toxicity to C. reinhardtii, mainly due to light blockage resulting from the PSNPs-microalgae heteroaggregation and the shading effect of PSNPs, which was independent of PSNPs concentrations. However, the toxicity of small-sized PSNPs (50 nm) was controlled by both particle surface charge and particle concentration. The positively charged PS-NH₂ was more readily heteroaggregated with microalgae than the negatively charged PS-COOH, leading to photosynthesis damage-induced toxicity. Increasing the concentration of small-sized PSNPs stimulated the secretion of extracellular polymeric substances, allowing more PSNPs to attach on the cell surface and further to enter the cell, which was responsible for the increased toxicity. These findings provide new insights into how nanoplastics induce contact toxicity in microalgae cells through specific biointerfacial interactions.