Triaxial bioprinting large-size vascularized constructs with nutrient channels

Abstract Bioprinting has demonstrated great advantages in tissue and organ regeneration. However, constructing large-scale tissue and organs in vitro is still a huge challenge due to the lack of some strategies for loading multiple types of cells precisely while maintaining nutrient channels. Here, a new 3D bioprinting strategy was proposed to construct large-scale vascularized tissue. A mixture of gelatin methacrylate (GelMA) and sodium alginate (Alg) was used as a bioink, serving as the outer and middle layers of a single filament in the triaxial printing process, and loaded with human bone marrow mesenchymal stem cells and human umbilical vein endothelial cells, respectively, while a calcium chloride (CaCl 2 ) solution was used as the inner layer. The CaCl 2 solution crosslinked with the middle layer bioink during the printing process to form and maintain hollow nutrient channels, then a stable large-scale construct was obtained through photopolymerization and ion crosslinking after printing. The feasibility of this strategy was verified by investigating the properties of the bioink and construct, and the biological performance of the vascularized construct. The results showed that a mixture of 5% (w/v) GelMA and 1% (w/v) Alg bioink could be printed at room temperature with good printability and perfusion capacity. Then, the construct with and without channels was fabricated and characterized, and the results revealed that the construct with channels had a similar degradation profile to that without channels, but lower compressive modulus and higher swelling rate. Biological investigation showed that the construct with channels was more favorable for cell survival, proliferation, diffusion, migration, and vascular network formation. In summary, it was demonstrated that constructing large-scale vascularized tissue by triaxial printing that can precisely encapsulate multiple types of cells and form nutrient channels simultaneously was feasible, and this technology could be used to prepare large-scale vascularized constructs.