Bioenergetic effects of pristine and ultraviolet-weathered polydisperse polyethylene terephthalate and polystyrene nanoplastics on human intestinal Caco-2 cells

The ubiquitous human exposure to nanoplastics (NPs) increasingly raises concerns regarding impact on our health. However, little is known on the biological effects of complex mixtures of weathered NPs with heterogenous size and irregular shape present in the environment. In this study, the bioenergetic effects of four such NPs mixtures on human intestinal Caco-2 cells were investigated. To this aim, Caco-2 cells were exposed to polydisperse nanoPET (<800 nm) and nanoPS (mixture of 100 and 750 nm) samples with and without ultraviolet (UV) weathering at low concentration range (10-10 particles/mL) for 48 h. Mitochondrial respiration, glycolytic functions and ATP production rates of exposed cells were measured by Seahorse XFe96 Analyzer. Among four NPs samples, polydisperse nanoPET with irregular shapes induced significant stimulation of mitochondrial respiration, glycolysis and ATP production rates in Caco-2 cells. Spherical nanoPS caused significant stimulation on glycolytic functions of Caco-2 cells at the highest concentration used (10 particles/mL). ATR-FTIR spectra and carbonyl index indicated formation of carbonyl groups in nanoPET and nanoPS after UV weathering. UV weathering could alleviate bioenergetic stress caused by NPs in Caco-2 cells and even shifted the energy pathways from mitochondrial respiration to glycolysis due to electrostatic repulsion between negatively charged UV-aged NPs and cell membranes. This research is the first to study in-vitro bioenergetic responses of NPs samples with multidimensional features (polymer type, irregular shape, heterogenous size, UV-weathering) on human health. It highlights that effects between pristine and weathered NPs are different at a bioenergetic level, which has important implications for the risk assessment of NPs on human health.