Molecular mechanism for combined toxicity of micro(nano)plastics and carbon nanofibers to freshwater microalgae Chlorella pyrenoidosa

The understanding of the environmental consequences resulting from the presence of micro(nano)plastics and carbon nanofibers (CNFs) in aquatic ecosystems is currently limited. This research endeavor sought to investigate the underlying molecular mechanisms by which engineered polystyrene-based microplastics (MPs)/nanoplastics (NPs) and CNFs, both individually and in combination, elicit toxic effects on an algal species Chlorella pyrenoidosa. The findings revealed that the combined toxicity of MPs/NPs and CNFs depended on the concentration of the mixture. As the concentration increased, the combined toxicity of MPs/NPs and CNFs was significantly greater than the toxicity of each component on its own. Furthermore, the combined toxicity of NPs and CNFs was higher than that of MPs and CNFs. The study integrated data on cell membrane integrity, oxidative stress, and antioxidant modulation to create an Integrated Biomarker Response index, which demonstrated that the co-exposure of algae to NPs and CNFs resulted in more severe cellular stress compared to exposure to NPs alone. Similarly, the combination of NPs and CNFs caused greater cellular stress than the combination of MPs and CNFs. Additionally, significant changes in the expression of stress-related genes caused by MPs/NPs alone and in combination with CNFs indicated that oxidative stress response, glucose metabolism, and energy metabolism played critical roles in particle-induced toxicity. Overall, this study provides the first insight into the toxicological mechanism of MPs/NPs and CNFs mixtures at the molecular level in freshwater microalgae.