Rapid Fabrication of Ready-to-Use Gelatin Scaffolds with Prevascular Networks Using Alginate Hollow Fibers as Sacrificial Templates

In this study, we developed a facile manufacturing method for interconnected prevascular networks using calcium chloride (CaCl₂) cross-linked alginate hollow fibers as sacrificial templates. The resulting network can be used to deliver oxygen and nutrients and remove waste for embedded cells in large-volume gelatin scaffolds during in vitro culturing. The sacrificial templates were printed by customized coaxial nozzles and embedded in scaffolds made of a mixture of gelatin, microbial transglutaminase (mTG), and sodium citrate. During the cross-linking of gelatin and mTG, the sacrificial templates started to dissolve from the scaffold-template interface due to the presence of the sodium citrate in the gelatin. The embedded sacrificial templates were completely dissolved without any postprocessing, and the designed prevascular networks successfully retained their geometries and dimensions. No residue of the template was observed at the scaffold-template interface after dissolution, which promoted cell adhesion. This manufacturing method has a high degree of freedom in templates' geometry, which was demonstrated by fabricating prevascular networks with various designs, including grid, branched, and dendritic networks. The effects of hollow fiber size and sodium citrate concentration on the dissolution time were analyzed. Human umbilical vein endothelial cells were injected into the aforementioned networks and formed a confluent endothelial cell monolayer with high viability during the culture process. The results suggest great promise to rapidly build large-scale ready-to-use gelatin scaffolds with prevascular networks for the applications in tissue engineering.