3D-bioprinted cell-laden hydrogel with anti- inflammatory and anti-bacterial activities for tracheal cartilage regeneration and restoration

Despite the notable advances in tissue-engineered tracheal cartilage (TETC), there remain several challenges that need to be addressed, such as uneven cell distribution for cartilage formation, customized C-shaped tracheal morphology, local inflammatory reactions, and infections. To overcome these challenges, this study proposed the addition of icariin (ICA) and chitosan (CS) into a gelatin methacryloyl (GelMA) hydrogel to develop a new ICA/CS/GelMA hydrogel with anti-inflammatory and anti-bacterial properties, and three-dimensional (3D)-bioprinting feasibility. The aim of this study was to construct a TETC, a customized C-shaped cartilage structure, with uniform chondrocyte distribution as well as anti-inflammatory and anti-bacterial functions. Our results confirmed that ICA/CS/GelMA hydrogel provides desirable rheological properties, suitable printability, favorable biocompatibility, and simulated microenvironments for chondrogenesis. Moreover, the addition of ICA stimulated chondrocyte proliferation, extracellular matrix synthesis, and anti-inflammatory ability, while the encapsulation of CS enhanced the hydrogels' anti-bacterial ability. All these led to the formation of an enhanced TETC after submuscular implantation and an elevated survival rate of experimental rabbits after orthotopic tracheal transplantation. This study provides a reliable cell-laden hydrogel with antiinflammatory and antibacterial activities, suitable printability, and significant advancements in in vivo cartilage regeneration and in situ tracheal cartilage restoration.