Formulation of chitosan and chitosan-nanoHAp bioinks and investigation of printability with optimized bioprinting parameters

The development of a chitosan-based bioink that can provide a cell-friendly environment at relatively low concentration and moderate cross-linking conditions is still problematic. Here, we developed amorphous nanohydroxyapatite (nHAp) containing chitosan bioink formulations that can be gelled via the inclusion of glycerol phosphate (GP) and sodium hydrogen carbonate (SHC) into the polymer network under physiological conditions. Rheological analyses indicated that all the formulations showed shear-thinning characteristics compatible with the extrusion-based bioprinting. Also, the chitosan bioinks exhibited more gel-like structure as the weight fraction of nHAp increased from 10 % to 40 %. The printability of the chitosan-based bioinks was assessed and optimized by response surface methodology (RSM). These studies revealed that all the formulations can be successfully printed within the ranges of 50–70 kPa printing pressure and 4–11 mm/s printing speed. Multi-layered chitosan biomaterials with distinct pore structure were successfully fabricated with a high printability index. High cell viability was observed after bioprinting with pre-osteoblastic MC3T3-E1 cells. In conclusion, this study represents for the first time that chitosan biomaterials bearing suitable rheological properties and cellularity can be printed with controllable architecture for 3D bone scaffolds.