Preparation of fragmented polyethylene nanoplastics using a focused ultrasonic system and assessment of their cytotoxic effects on human cells

Abstract

With the growing prevalence of plastic use, the environmental release of plastic waste is escalating, and fragmented nanoscale plastic particles are emerging as significant environmental threats. This study aimed to evaluate the cytotoxic effects of fragmented polyethylene nanoplastics (PE NPs) manufactured using a focused ultrasonic system. The ultrasonic irradiation process generated fragmented PE NPs with a geometric mean diameter of 85.14 ± 5.37 nm and a size range of 25–350 nm. To assess cytotoxicity, we conducted a series of tests on various human cell lines, including stomach, blood, colon, lung, skin, liver, and brain-derived cells. The testing involved MTS-based cell viability assays to evaluate direct impacts on cell viability, lactate dehydrogenase (LDH) leakage assays to measure membrane damage, and ELISA to quantify TNF-α release as an indicator of inflammation. Although PE-NPs did not immediately induce apoptosis, significant LDH leakage and elevated TNF-α levels were observed across all cell lines, indicating membrane damage and inflammatory responses. Additionally, flow cytometry and TEM analyses revealed the intracellular accumulation of PE-NPs, further supporting their cytotoxic potential. These results demonstrate that fragmented PE-NPs can disrupt cellular membranes and induce inflammatory responses through accumulation within cells. The findings suggest that these NPs pose potential hazards to cell viability and underscore the need for further research into their environmental and health impacts.