Mixed ultrashort peptide bioinks for improved 3D bioprinting of self-healing trachea-like constructs

Concerns related to the trachea frequently arise from obstructive conditions and occlusions, such as tracheal stenosis, tracheomalacia, traumatic disruptions, and papillary thyroid carcinoma. These medical challenges underscore the need for new biomaterials to support tissue engineering for tissue regeneration. The advent of 3D bioprinting technology has emerged as a pivotal advancement, facilitating the fabrication of patient-specific, biocompatible, cell-laden constructs. This technological advancement enables the controlled promotion of cell growth and tissue development, thereby offering a promising avenue for tissue regeneration. In this study, we developed mixed ultrashort peptide bioinks for three-dimensional (3D) bioprinting of a trachea-like construct that exhibits self-healing and elastic properties. We employed a stiffness prediction map (SPM) as an empirical tool to predict the physical characteristics and stiffness behavior of the mixed bioinks, thereby facilitating the optimization of the 3D bioprinting process. The SPM enabled the fine-tuning of these bioinks by identifying peptide mixtures that successfully mimic the natural stiffness of the perichondral niche microenvironment. These mixed bioinks successfully promoted mesenchymal stromal cell differentiation towards chondrocyte formation, thereby facilitating the biofabrication of elastic 3D-printed structures for trachea regeneration. Our bioinks exhibited remarkable printing resolution and mechanical properties while supporting cell growth and chondrogenesis. A bioprinted trachea-like model, cultured for up to 100 days, showed excellent mechanical properties, resulting in a stable, yet elastic biomaterial. This study is the first to combine SPM with 3D bioprinting for the fabrication of a trachea-like model, supporting the development of advanced self-healing biomaterials for trachea tissue regeneration.