Solvent types used for the preparation of hydrogels determine their mechanical properties and influence cell viability through gelatine and calcium ions release.

Alginate-gelatin hydrogels are the most commonly used materials for 3D bioprinting. Their printability depends on their properties, and these derive from the way they are prepared and their very composition. Therefore, the aim of the study was to investigate the type of solvent (deionized water, phosphate buffer, and culture medium) and contents of gelatin in the composition of hydrogel (2% wt/vol alginate, 6% and 9% wt/vol of gelatin) on their biological, physicochemical, and mechanical properties, as well as printability and the ability of cells to proliferate in the printed structures. The results obtained revealed that all the manufactured hydrogel materials are biocompatible. The use of deionized water as a solvent results in the highest degree of cross-linking of hydrogels, thus obtaining a polymer with the highest rigidity. Moreover, an increase in gelatin content leads to an increase in the Young's modulus value, irrespectively of the solvent in which the hydrogels were prepared. Based on the chemical structure, it is more reasonable to use a culture medium for bioink preparation due to free NH and NH groups being present, which are ligands for cell attachment and their proliferation. For the selected material (2A9GM), the printability and high viability of the cells after printing were confirmed. In this case, the concentration of the cross-linking agent influences gelatin amount release and calcium ions release, and these two processes determine the change in the viability of the cells encapsulated in the bioink.