3D Printing of Shear-Thinning Hyaluronic Acid Hydrogels with Secondary Cross-Linking

The development of printable biomaterial inks is critical to the application of 3D printing in biomedicine. To print high-resolution structures with fidelity to a computer-aided design, materials used in 3D printing must be capable of being deposited on a surface and maintaining a printed structure. A dual-cross-linking hyaluronic acid system was studied here as a printable hydrogel ink, which encompassed both shear-thinning and self-healing behaviors via guest-host bonding, as well as covalent cross-linking for stabilization using photopolymerization. When either guest-host assembly or covalent cross-linking was used alone, long-term stable structures were not formed, because of network relaxation after printing or dispersion of the ink filaments prior to stabilization, respectively. The dual-cross-linking hydrogel filaments formed structures with greater than 16 layers that were stable over a month with no loss in mechanical properties and the printed filament size ranged from 100 to 500 μm, depending on printing parameters (needle size, speed, and extrusion flux). Printed structures were further functionalized (i.e., RGD peptide) to support cell adhesion. This work highlights the importance of ink formulation and cross-linking on the printing of stable hydrogel structures.