Cutting-edge 3D COPD lung mimic

<bold>Introduction:</bold> Chronic obstructive pulmonary disease (COPD), the third leading global cause of death, lacks innovative therapeutics due to the absence of efficient lung models. This study presents an in vitro lung mimic for healthy and COPD conditions. <bold>Methods:</bold> (1) Employing CNBio's PhysioMimix, we developed a microfluidic 3D system to emulate dynamic human lung flow conditions, ensuring unidirectional cell flow throughout the culture. (2) Recognizing extracellular matrix (ECM)'s influence on cellular activity, we employed collagen type I and fibronectin to functionalize substrates. (3) Comparing co-cultures of normal human lung fibroblasts (NHLFs) with either normal or diseased human bronchial epithelial cells (NHBE, DHBE) created healthy and COPD lung mimics. To assess model reproducibility, we implemented a comprehensive QC strategy involving real-time and end-point measurements, including light microscopy, scanning electron microscopy (SEM), impedance spectroscopy with transepithelial electrical resistance (TEER), immunofluorescence (IF) staining, and cytokine profiling. <bold>Results:</bold> (1) Dynamic conditions showcased increased phenotype heterogeneity (light microscopy) and hairy-like ciliated cell structures (SEM), indicating enhanced barrier protection and mucociliary clearance. (2) Impedance spectroscopy revealed collagen-I's superior impact on cell adhesion, growth, and differentiation compared to fibronectin. (3) DHBEs exhibited compromised barrier integrity (TEER), goblet cell hypertrophy (IF), reduced cilia (SEM), and an inflammatory phenotype (cytokine). <bold>Conclusion:</bold> Our study emphasizes technological considerations in constructing lung models. We developed a representative COPD model with comprehensive QC, facilitating COPD therapeutic discovery.