Construction and Application of in vitro Alveolar Models Based on 3D Printing Technology

Increasing lung diseases, mutating coronaviruses, and the development of new compounds urgently require biomimetic in vitro lung models for lung pathology, toxicology, and pharmacology. The current construction strategies for lung models mainly include animal models, 2D cell culture, lung-on-a-chip, and lung organoids. However, current models face difficulties in reproducing in vivo-like alveolar size and vesicle-like structures, and are unable to contain multiple cell types. In this study, a strategy for constructing alveolar models based on degradable hydrogel microspheres is proposed. Hydrogel microspheres, 200–250 µm in diameter, were prepared using a self-developed printing technique driven by alternating viscous and inertial forces. Microcapsules were further constructed using a coacervation-based layer-by-layer technique and core liquefaction. Three types of cells were inoculated and co-cultured on hydrogel capsules based on optimized microcapsule surface treatment strategies. Finally, an in vitro three-dimensional endothelial alveolar model with a multicellular composition and vesicle-like structure with a diameter of approximately 230 µm was successfully constructed. Cells in the constructed alveolar model maintained a high survival rate. The LD₅₀ values of glutaraldehyde based on the constructed models were in good agreement with the reference values, validating the potential of the model for future toxicant and drug detection.