Three-dimensional printing of cell-laden microporous constructs using blended bioinks.

Abstract

Hydrogels such as alginate and gelatin have shown potential as biomaterials in various three-dimensional (3D) bioprinting applications. However, parameters such as viscosity, porosity, and printability influence the performance of hydrogel-based biomaterials, and there are limited characterization studies conducted on the behavior of these constructs. In this work, a syringe-based extrusion bioprinter was used to print 3D constructs with bioink composed of various concentrations of alginate and gelatin along with fibrinogen and human umbilical vein endothelial cells. Instead of crosslinking the gelatin, the gelatin was left uncrosslinked to provide microporosity within the system that can impact the cellular response. Mechanical and biochemical characterization was performed to evaluate the structural stability and integrity of the printed constructs along with viability of embedded cells. Bioprinted constructs of a higher total concentration of alginate and gelatin yielded better stability and structural integrity after culture. More importantly, higher amounts of gelatin (i.e., 1:9 instead of 2:3 alginate:gelatin) were shown to improve printability, which is different than most studies that instead use alginate to improve printability. In addition, higher amounts of gelatin impacted the changes in surface morphological features of the constructs after incubation, and ultimately improved biocompatibility with our system. Overall, this study demonstrated that an uncrosslinked gelatin system can provide flexible printing parameters and surface morphologies, but careful control over the printing parameters may be required. The bioink concentration of 10% (w/v) with minimum alginate and higher gelatin concentration exhibited the best printability, cell survival, and viability.