Low-temperature 3D printing of collagen and chitosan composite for tissue engineering

Highlights: • A series of hybrid collagen/chitosan bioinks were prepared with good printability. • The bioinks were successfully printed through careful temperature controlling. • The 3D scaffolds had delicate structure, regulated physiochemical properties and high cell viability. Three-dimensional (3D) printing is a promising method to prepare scaffolds for tissue regeneration. Collagen and chitosan composites are superior materials for tissue engineering scaffold but rarely printed due to their poor printability. Here, we prepared a series of tunable hybrid collagen/chitosan bioinks with significantly improved printability through hydrogen bond interaction and printed them into scaffolds by carefully controlling the temperature. Rheological tests proved the printable bioinks had sound shear thinning behavior, dramatical viscosity variation with temperature, and the gelation temperature from 7 to 10 °C. Chitosan could decrease the swelling ratio of the printed scaffolds, while their degradation rate increased with collagen proportion and the values of Young's modulus and tensile strength increased with chitosan proportion. Moreover, the scaffolds containing 2% (m/v) collagen and 2% (m/v) chitosan had a homogeneous and compact honeycomb-like structure, demonstrating the strengthening effect of chitosan. Cell viability assay presented vigorous cell growth on the surface of scaffolds, meanwhile, live cells were also found inside and at the bottom of the scaffolds, indicating the migration of cells. Therefore, chitosan can improve the printability of collagen and the hybrid collagen/chitosan bioinks can be printed into scaffolds with regulated properties, thus can fit different applications in tissue engineering.