DOI: 10.1038/s41598-025-00438-z
Arunima Sengupta 1 2, Saskia Schmid 3 4, Noémie Grangier 4, Aurélien Dorn 3 4, Marco Hebestreit 5, Andreas Hugi 6, Kristína Žajdlíková 7 8, Anja Herbst 9 10, Paula Losada-Oliva 11, Heidi Ortolf-Wahl 5, Philippe Krebs 9, Janick D Stucki 6, Vera van der Velpen 7 8, Jesus Perez-Gil 11, Tobias Krebs 5, Nina Hobi 6, Olivier T Guenat 12
1Organs-On-Chip Technologies, ARTORG Center for Biomedical Engineering, University of Bern, Bern, Switzerland.
2Alexis Technologies AG, Bern, Switzerland.
3Organs-On-Chip Technologies, ARTORG Center for Biomedical Engineering, University of Bern, Bern, Switzerland.
4Alexis Technologies AG, Bern, Switzerland.
5VITROCELL Systems GmbH, Waldkirch, Germany.
6Swiss Organs-On-Chip Innovation, AlveoliX AG, Bern, Switzerland.
7Clinical Pharmacology and Toxicology, Department of General Internal Medicine, Inselspital, Bern University Hospital, University of Bern, Bern, Switzerland.
8Institute of Pharmacology, University of Bern, Bern, Switzerland.
9Institute of Tissue Medicine and Pathology, University of Bern, Bern, Switzerland.
10Graduate School for Cellular and Biomedical Sciences, University of Bern, Bern, Switzerland.
11Department of Biochemistry, Faculty of Biology, and Research Institute “Hospital 12 de Octubre (i+12)”, Universidad Complutense, Madrid, Spain.
12Organs-On-Chip Technologies, ARTORG Center for Biomedical Engineering, University of Bern, Bern, Switzerland.
Abstract
Continuous exposure to cigarette smoke (CS) significantly contributes to the development and progression of chronic obstructive pulmonary disease (COPD) and lung cancer. Animal models that inhale smoke nasally and have different lung physiology from humans may not accurately replicate cigarette smoke-induced health effects. Furthermore, traditional in vitro models fail to replicate the lung’s dynamic mechanical forces and realistic inhalation exposure patterns, limiting their relevance in preclinical research. Here, we introduce an advanced smoke inhalation-based lung-on-chip system, the Continuous Flow AX12 (CFAX12), to investigate CS-induced cellular responses in a physiologically relevant manner. Unlike previous technologies, the CFAX12 integrates cyclic mechanical stretch with controlled whole-smoke exposure, allowing for a more accurate recreation of CS-induced alveolar microenvironment dynamics and barrier integrity responses. Using human alveolar epithelial cells, lung microvascular endothelial cells, and macrophages in mono- and co-culture models under air-liquid interface (ALI) conditions with breathing-like stretch (Str), we simulated key lung microenvironment features. Our results show that CS exposure using the CFAX12 induced a ~ 60% reduction in trans-barrier electrical resistance (TER), increased ROS generation depending on cellular model complexity, and a ~ 4.5-fold increase in IL-8 gene expression, all key hallmarks of early COPD pathogenesis. These findings underscore smoke-induced epithelial damage, inflammation, and oxidative stress, all of which contribute to alveolar barrier dysfunction and disease progression. Also, CFAX12 provides a more physiologically relevant alternative to submerged cigarette smoke extract (CSE) treatments, offering controlled whole-smoke exposure using the VC10 Smoking Robot, ensuring precisely regulated smoke delivery. Additionally, inclusion of pulmonary surfactant reduced IL8 gene levels by ~ 5 folds. Hence, by integrating mechanical and biological complexity, CFAX12 offers a robust platform for assessing inhaled smoke effects and identifying therapeutic targets. It’s application in COPD drug screening can facilitate the discovery of compounds that preserve alveolar integrity, reduce inflammation, and mitigate oxidative damage, ultimately bridging the gap between regulatory and preclinical research applications.
