RESUMO
Progress in biofabrication technologies is mainly hampered by the limited number of suitable hydrogels that can act as bioinks. Here, we present a new bioink for 3D-printing, capable of forming large, highly defined constructs. Hydrogel formulations consisted of a thermoresponsive polymer mixed with a poly(ethylene glycol) (PEG) or a hyaluronic acid (HA) cross-linker with a total polymer concentration of 11.3 and 9.1 wt% respectively. These polymer solutions were partially cross-linked before plotting by a chemoselective reaction called oxo-ester mediated native chemical ligation, yielding printable formulations. Deposition on a heated plate of 37 °C resulted in the stabilization of the construct due to the thermosensitive nature of the hydrogel. Subsequently, further chemical cross-linking of the hydrogel precursors proceeded after extrusion to form mechanically stable hydrogels that exhibited a storage modulus of 9 kPa after 3 hours. Flow and elastic properties of the polymer solutions and hydrogels were analyzed under similar conditions to those used during the 3D-printing process. These experiments showed the ability to extrude the hydrogels, as well as their rapid recovery after applied shear forces. Hydrogels were printed in grid-like structures, hollow cones and a model representing a femoral condyle, with a porosity of 48 ± 2%. Furthermore, an N-hydroxysuccinimide functionalized thermoplastic poly-ε-caprolactone (PCL) derivative was successfully synthesized and 3D-printed. We demonstrated that covalent grafting of the developed hydrogel to the thermoplastic reinforced network resulted in improved mechanical properties and yielded high construct integrity. Reinforced constructs also containing hyaluronic acid showed high cell viability of chondrocytes, underlining their potential for further use in regenerative medicine applications.
