The present work describes the implementation of an in-vitro model for the evaluation of the injury mechanisms of lung epithelial cells during simulated airway reopening events. Collagen-coated polyacrylamide substrates with different rigidities served as cell culture substrates, and a parallel plate perfusion chamber was used for simulating the reopening of fluid-filled airways. Human alveolar epithelial cell (A549) monolayers were grown to confluence on the different substrates and the cellular response was evaluated in terms of cytoskeletal distribution, proliferation rate, and cell stiffness. One and five reopening events were simulated, and cell response was characterized via live/dead fluorescence imaging. Cells cultured on stiffer substrates showed higher proliferation rate, as well as a more spread and tightly packed morphology. Airway reopening simulations showed that cells cultured on softer substrates are less susceptible to cell injury.

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