Rheological characterization of cell-laden alginate-gelatin hydrogels for 3D biofabrication

Biofabrication of tissue models that closely mimic the tumor microenvironment is necessary for high-throughput anticancer therapeutics. Extrusion-based bioprinting of heterogeneous cell-laden hydrogels has shown promise in advancing rapid artificial tissue development. A major bottleneck limiting the rapid production of physiologically relevant tissue models is the current limitation in effectively printing large populations of cells. However, by significantly increasing hydrogel cell-seeding densities, the time required to produce tissues could be effectively reduced. Here, we explore the effects of increasing cell seeding densities on the viscoelastic properties, printability, and cell viability of two different alginate-gelatin hydrogel compositions. Rheological analysis of hydrogels of varying cell seeding densities reveals an inverse relationship between cell concentration and zero-shear viscosity. We also observe that as cell seeding densities increases, the storage moduli decrease, thus lowering the required printing pressures for gel extrusion. We also observe that increasing cell concentration can negatively impact the structural properties of the extruded material by increasing post-print line spreading. We find that hydrogels composed of higher molecular weight alginates and the highest cell-seeding densities (10⁷ cells/mL) yield higher cell viability (>80%) and structural uniformity after printing. The optimized printing parameters determined for the alginate-gelatin bioinks explored may aid in the future rapid fabrication of functional tissue models for therapeutic screening.