Chlorine disinfection elevates the toxicity of polystyrene microplastics to human cells by inducing mitochondria-dependent apoptosis

Microplastics (MPs) are ubiquitous in drinking water and pose potential threats to human health. Despite increasingly attentions on the toxicity of MPs, the deleterious effects of MPs after chlorine disinfection, which might be a more accessible form of MPs, has rarely been considered. Here, we first treated pristine polystyrene microplastics (PS-MPs) with chlorine to simulate the reactions that occur during drinking water treatment, and investigated and compared the cytotoxicity of chlorinated PS-MPs to those of pristine PS-MPs. Chlorine disinfection did not change the size of pristine PS-MPs, but increased the surface roughness. In addition, abundant carbon-chlorine bonds and persistent free radicals were generated on the surface of chlorinated PS-MPs. Compared with pristine PS-MPs, chlorinated PS-MPs markedly inhibited the cell proliferation, changed cellular morphology, destroyed cell membrane integrity, induced cell inflammatory response and apoptosis. Proteomics confirmed the difference in interactions with intracellular proteins between these particles. Furthermore, we found that the regulation of PI3K/AKT and Bcl-2/Bax pathways, oxidative stress-triggered mitochondrial depolarization, and the activation of caspase cascade were identified as the underlying mechanisms for the enhanced apoptosis ratio in GES-1 cells when exposed to chlorinated PS-MPs. This exacerbated cytotoxicity could be explained by the enhanced surface roughness and changed surface chemistry of these PS-MPs after chlorine disinfection. This work discloses the impacts of chlorine disinfection on the cytotoxicity of PS-MPs, which provides new insights for a more systematic risk assessment of MPs.