Pulmonary hazards of nanoplastic particles: a study using polystyrene in in vitro models of the alveolar and bronchial epithelium

Abstract Background Nanoplastics (NPs) are released into the environment through the degradation of plastic objects, leading to human exposure. Due to their small size, concerns have been raised about the potential hazards to the respiratory tract, as ultrafine and nanoparticles are known to penetra...

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Main Authors: Sara Michelini, Safaa Mawas, Ema Kurešepi, Francesco Barbero, Katarina Šimunović, Dorian Miremont, Stéphanie Devineau, Martin Schicht, Victor Ganin, Øyvind Pernell Haugen, Anani Komlavi Afanou, Charlotte Izabelle, Shan Zienolddiny-Narui, Katharina Jüngert, Neža Repar, Ivana Fenoglio, Barbara Šetina Batić, Friedrich Paulsen, Ines Mandić-Mulec, Sonja Boland, Andreja Erman, Damjana Drobne
Format: Article
Language:English
Published: BMC 2025-05-01
Series:Journal of Nanobiotechnology
Online Access:https://doi.org/10.1186/s12951-025-03419-6
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author Sara Michelini
Safaa Mawas
Ema Kurešepi
Francesco Barbero
Katarina Šimunović
Dorian Miremont
Stéphanie Devineau
Martin Schicht
Victor Ganin
Øyvind Pernell Haugen
Anani Komlavi Afanou
Charlotte Izabelle
Shan Zienolddiny-Narui
Katharina Jüngert
Neža Repar
Ivana Fenoglio
Barbara Šetina Batić
Friedrich Paulsen
Ines Mandić-Mulec
Sonja Boland
Andreja Erman
Damjana Drobne
author_facet Sara Michelini
Safaa Mawas
Ema Kurešepi
Francesco Barbero
Katarina Šimunović
Dorian Miremont
Stéphanie Devineau
Martin Schicht
Victor Ganin
Øyvind Pernell Haugen
Anani Komlavi Afanou
Charlotte Izabelle
Shan Zienolddiny-Narui
Katharina Jüngert
Neža Repar
Ivana Fenoglio
Barbara Šetina Batić
Friedrich Paulsen
Ines Mandić-Mulec
Sonja Boland
Andreja Erman
Damjana Drobne
author_sort Sara Michelini
collection DOAJ
description Abstract Background Nanoplastics (NPs) are released into the environment through the degradation of plastic objects, leading to human exposure. Due to their small size, concerns have been raised about the potential hazards to the respiratory tract, as ultrafine and nanoparticles are known to penetrate till the alveolar regions of the lungs, potentially impairing their functions. Thus, in the present study, we used model polystyrene nanoparticles doped with the fluorescent metal europium (PS-Eu) to enhance the understanding of NPs hazard and investigate adverse outcomes associated with exposure in human lungs using alveolar (A549) and bronchial (Calu-3) cell models grown in 2D and 3D submerged conditions or quasi air-liquid interface (ALI) conditions (3D). Results Briefly, after in-dept physicochemical characterization of the particles, we assessed their impact on ROS production, cell viability (AlamarBlue and lactate dehydrogenase assays) and barrier integrity (lucifer yellow assay and TEER measurement), finding no negative effects in either model. However, in alveolar cells, particles increased acidic organelle activity. Transmission electron microscopy and Raman microscopy showed, in both models, a dose- and cell-dependent particle uptake with PS-Eu accumulating in numerous and large endo-lysosomes, which, in transwells-grown A549 cells, often contained also lamellar bodies (LBs), organelles involved in surfactants storage and secretion. After extensively quantifying surfactant proteins (SP) in the pellet and supernatant fractions of treated A549 cells, we observed a significant reduction in several members of this family, including surfactant protein B, which is crucial for lamellar body formation and surface tension regulation in the lungs. In quasi-ALI Calu-3 cultures instead, PS-Eu significantly upregulated interleukin 6 (IL-6) and increased transforming growth factor beta β (TGF-β), zonula occludens 1 (ZO-1), and mucin (MUC) 5B mRNA expressions causing a moderate proinflammatory response. Conclusion Our results show that PS-Eu exposure does not induce acute cytotoxicity in these models, but affects cell-specific functions like surfactant, mucin, and cytokine production. This underscores the limitations of relying solely on standard cytotoxicity tests for particle hazard assessment and highlights the importance of investigating cell function-specific signaling pathways. To support researchers in hazard assessment, we propose specific classes of biomarkers to test in in vitro lung models.
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spelling doaj-art-3fe8b639393d4928a33f048998d3e3372025-08-20T03:22:03ZengBMCJournal of Nanobiotechnology1477-31552025-05-0123113410.1186/s12951-025-03419-6Pulmonary hazards of nanoplastic particles: a study using polystyrene in in vitro models of the alveolar and bronchial epitheliumSara Michelini0Safaa Mawas1Ema Kurešepi2Francesco Barbero3Katarina Šimunović4Dorian Miremont5Stéphanie Devineau6Martin Schicht7Victor Ganin8Øyvind Pernell Haugen9Anani Komlavi Afanou10Charlotte Izabelle11Shan Zienolddiny-Narui12Katharina Jüngert13Neža Repar14Ivana Fenoglio15Barbara Šetina Batić16Friedrich Paulsen17Ines Mandić-Mulec18Sonja Boland19Andreja Erman20Damjana Drobne21Biotechnical Faculty, Department of Biology, University of LjubljanaUniversité Paris Cité, CNRS, Unité de Biologie Fonctionnelle et AdaptativeBiotechnical Faculty, Department of Biology, University of LjubljanaDepartment of Chemistry, Laboratory of Toxicity and Biocompatibility of Materials, University of TorinoBiotechnical Faculty, Department of Microbiology, University of LjubljanaUniversité Paris Cité, CNRS, Unité de Biologie Fonctionnelle et AdaptativeUniversité Paris Cité, CNRS, Unité de Biologie Fonctionnelle et AdaptativeInstitute of Functional and Clinical Anatomy, Friedrich-Alexander-University of Erlangen-NürnbergInstitute of Metals and TechnologySTAMI, National Institute of Occupational HealthSTAMI, National Institute of Occupational HealthUniversité Paris Cité, CNRS UAR612, Inserm US25, Cellular and Molecular Imaging FacilitySTAMI, National Institute of Occupational HealthInstitute of Functional and Clinical Anatomy, Friedrich-Alexander-University of Erlangen-NürnbergBiotechnical Faculty, Department of Biology, University of LjubljanaDepartment of Chemistry, Laboratory of Toxicity and Biocompatibility of Materials, University of TorinoInstitute of Metals and TechnologyInstitute of Functional and Clinical Anatomy, Friedrich-Alexander-University of Erlangen-NürnbergBiotechnical Faculty, Department of Microbiology, University of LjubljanaUniversité Paris Cité, CNRS, Unité de Biologie Fonctionnelle et AdaptativeFaculty of Medicine, Institute of Cell Biology, University of LjubljanaBiotechnical Faculty, Department of Biology, University of LjubljanaAbstract Background Nanoplastics (NPs) are released into the environment through the degradation of plastic objects, leading to human exposure. Due to their small size, concerns have been raised about the potential hazards to the respiratory tract, as ultrafine and nanoparticles are known to penetrate till the alveolar regions of the lungs, potentially impairing their functions. Thus, in the present study, we used model polystyrene nanoparticles doped with the fluorescent metal europium (PS-Eu) to enhance the understanding of NPs hazard and investigate adverse outcomes associated with exposure in human lungs using alveolar (A549) and bronchial (Calu-3) cell models grown in 2D and 3D submerged conditions or quasi air-liquid interface (ALI) conditions (3D). Results Briefly, after in-dept physicochemical characterization of the particles, we assessed their impact on ROS production, cell viability (AlamarBlue and lactate dehydrogenase assays) and barrier integrity (lucifer yellow assay and TEER measurement), finding no negative effects in either model. However, in alveolar cells, particles increased acidic organelle activity. Transmission electron microscopy and Raman microscopy showed, in both models, a dose- and cell-dependent particle uptake with PS-Eu accumulating in numerous and large endo-lysosomes, which, in transwells-grown A549 cells, often contained also lamellar bodies (LBs), organelles involved in surfactants storage and secretion. After extensively quantifying surfactant proteins (SP) in the pellet and supernatant fractions of treated A549 cells, we observed a significant reduction in several members of this family, including surfactant protein B, which is crucial for lamellar body formation and surface tension regulation in the lungs. In quasi-ALI Calu-3 cultures instead, PS-Eu significantly upregulated interleukin 6 (IL-6) and increased transforming growth factor beta β (TGF-β), zonula occludens 1 (ZO-1), and mucin (MUC) 5B mRNA expressions causing a moderate proinflammatory response. Conclusion Our results show that PS-Eu exposure does not induce acute cytotoxicity in these models, but affects cell-specific functions like surfactant, mucin, and cytokine production. This underscores the limitations of relying solely on standard cytotoxicity tests for particle hazard assessment and highlights the importance of investigating cell function-specific signaling pathways. To support researchers in hazard assessment, we propose specific classes of biomarkers to test in in vitro lung models.https://doi.org/10.1186/s12951-025-03419-6
spellingShingle Sara Michelini
Safaa Mawas
Ema Kurešepi
Francesco Barbero
Katarina Šimunović
Dorian Miremont
Stéphanie Devineau
Martin Schicht
Victor Ganin
Øyvind Pernell Haugen
Anani Komlavi Afanou
Charlotte Izabelle
Shan Zienolddiny-Narui
Katharina Jüngert
Neža Repar
Ivana Fenoglio
Barbara Šetina Batić
Friedrich Paulsen
Ines Mandić-Mulec
Sonja Boland
Andreja Erman
Damjana Drobne
Pulmonary hazards of nanoplastic particles: a study using polystyrene in in vitro models of the alveolar and bronchial epithelium
Journal of Nanobiotechnology
title Pulmonary hazards of nanoplastic particles: a study using polystyrene in in vitro models of the alveolar and bronchial epithelium
title_full Pulmonary hazards of nanoplastic particles: a study using polystyrene in in vitro models of the alveolar and bronchial epithelium
title_fullStr Pulmonary hazards of nanoplastic particles: a study using polystyrene in in vitro models of the alveolar and bronchial epithelium
title_full_unstemmed Pulmonary hazards of nanoplastic particles: a study using polystyrene in in vitro models of the alveolar and bronchial epithelium
title_short Pulmonary hazards of nanoplastic particles: a study using polystyrene in in vitro models of the alveolar and bronchial epithelium
title_sort pulmonary hazards of nanoplastic particles a study using polystyrene in in vitro models of the alveolar and bronchial epithelium
url https://doi.org/10.1186/s12951-025-03419-6
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