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Polystyrene nanoplastics impair epithelial barrier establishment and modulate extracellular vesicle release in human bronchial cells cultured at the air-liquid interface

Toxicology 2026
Cristina Milillo, Annalisa Bruno, Francesca D’Ascanio, Marco Gatta, Michela Battistelli, Sabrina Burattini, Piero Di Carlo, Jorge Parrón‐Ballesteros, Elena Niccolai, Melania Dovizio, Paola Lanuti, Eva Batanero, Amedeo Amedei, Patrizia Ballerini

Summary

Scientists exposed lab-grown human airway cells to tiny nanoplastic particles (like those found in breathing air polluted with plastic waste) and found the particles weakened the protective barrier these cells normally form, while also triggering inflammation-related signals. This suggests nanoplastics could make airways more vulnerable, especially in people whose lung lining is already damaged or still developing—though this was shown in a lab model, not in humans breathing polluted air, so more research is needed to confirm real-world impact.

The growing production of plastics has led to widespread environmental contamination by micro- and nanoplastics (MNPLs), raising increasing concern about their potential impact on human respiratory health. Among them, nanoplastics (NPs) can penetrate deeply into the lungs. However, their impact on human bronchial epithelial cells remains poorly characterized. Here, we assessed the effects of 40 nm polystyrene (PS)-NPs on human bronchial Calu-3 cells cultured at the air-liquid interface (ALI) during early epithelial establishment, a three-dimensional model representing a functionally immature or compromised airway epithelial barrier. Chronic exposure (120 h) to PS-NPs (12 and 120 µg/cm²) interfered with barrier establishment in a concentration- and time-dependent manner, as demonstrated by reduced transepithelial electrical resistance (TEER) and decreased expression of the junctional proteins E-cadherin and Zonula Occludens-1 (ZO-1). PS-NPs also promoted the release of multiple inflammatory mediators, with osteopontin (OPN), a recognized biomarker of air pollution exposure, together with IL-2 and IL-10, showing the most consistent increases across both epithelial compartments. Other cytokines displayed concentration- and compartment-dependent secretion patterns. This polarized secretion pattern suggests selective activation of epithelial signaling pathways and paracrine communication toward the submucosal milieu. Furthermore, PS-NPs were internalized by Calu-3 cells and packaged into extracellular vesicles (EVs), a process that may limit acute cellular damage while potentially enabling propagation of NP-induced signals. Overall, these findings indicate that PS-NP exposure perturbs epithelial barrier function and polarized intercellular communication under conditions mimicking a compromised airway epithelium, suggesting a mechanistic role for NPs as aggravating factors of pre-existing epithelial dysfunction.

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