Three-dimensional bioprinting of polysaccharide-based self-healing hydrogels with dual cross-linking

Three-dimensional (3D) bioprinting technique is useful to fabricate constructs with functional and biological structures for various biomedical applications. Oxidized hyaluronate (OHA) and glycol chitosan (GC) can form autonomous self-healing hydrogels when adipic acid dihydrazide (ADH) is used. We demonstrate that hyaluronate-alginate hybrid (HAH) polymers can be used for secondary physical cross-linking of OHA/GC/ADH hydrogel with calcium ions after 3D printing. The molecular weight of hyaluronate can be varied while keeping the molecular weight of alginate in HAH. The mechanical stiffness and stability of gels after 3D printing are strongly dependent on the molecular weight of HAH at the same cross-linking density. In vitro chondrogenic differentiation of ATDC5 cells encapsulated in 3D-printed constructs is dependent on the molecular weight of HAH in gels. This dual cross-linking system consisting of naturally occurring biocompatible polysaccharides may have potential in the 3D bioprinting of custom-made scaffolds for tissue engineering applications.