Hybrid Biofabrication of Multilayered, Self-Supporting Tracheal Constructs Using Functionalized dECM Hydrogels

The trachea plays a vital role in maintaining airway patency and facilitating gas exchange. However, severe tracheal defects result from trauma, infection, congenital abnormalities, or malignancies. There are significant challenges with current treatments, such as synthetic implants, allografts, and autografts, suffering from immune rejection, donor shortages, poor mechanical integration, and inadequate long-term functionality. Existing tissue-engineered tracheal grafts often fail to replicate the native multilayered heterogeneous tissue structure and biomechanical properties, limiting their clinical success. To address this, we propose a multilayered heterogeneous tracheal construct using decellularized extracellular matrix (dECM) hydrogels by a hybrid approach of integrating digital light processing printing and gel casting, designed to mimic the native tracheal architecture and enhance cellular functionality. By this approach, we developed a self-standing multilayered tracheal construct by sequentially assembling cartilage, submucosa, and muscle layers using the dECM hydrogels functionalized with cells to replicate the native tracheal architecture. Structural integrity, ECM remodeling, and contractility were achieved within each layer. The developed multilayered tracheal construct demonstrated structural integrity and mechanical properties nearly comparable to native tracheal tissue, where it withstands airway dynamics along with cell viability, proliferation, and integration within each layer. Our hybrid approach of developing a multilayered tracheal construct presents a promising solution for tracheal reconstruction and regeneration, revolutionizing tracheal repair and improving treatment modalities.