Hydrogels tested and evaluated in this study were developed for the possibility of their use as the bioinks for 3D direct bioprinting. Procedures for preparation and sterilization of hydrogels and the speed of the bioprinting were developed. Sodium alginate gelatine hydrogels were characterized in terms of printability, mechanical, and biological properties (viability, proliferation ability, biocompatibility). A hydrogel with the best properties was selected to carry out direct bioprinting tests in order to determine the parameters of the bioink, adapted to print with use of the designed and constructed bioprinter and provide the best conditions for cell growth. The obtained results showed the ability to control mechanical properties, biological response, and degradation rate of hydrogels through the use of various solvents. The use of a dedicated culture medium as a solvent for the preparation of a bioink, containing the predicted cell line, increases the proliferation of these cells. Modification of the percentage of individual components of the hydrogel gives the possibility of a controlled degradation process, which, in the case of printing of temporary medical devices, is a very important parameter for the hydrogels’ usage possibility—both in terms of tissue engineering and printing of tissue elements replacement, implants, and organs.

The 3D printing technologies used for medical applications are mostly based on paste extruders. These are designed for high capacity, and thus often feature large material reservoirs and large diameter nozzles. A major challenge for most 3D printing platforms is a compromise between speed, accuracy, and/or volume/mass of moving elements. To address these issues, we integrated a peristaltic pump into a bioprinter. That allowed for combining the most important requirements: high precision, a large material reservoir, and safety of biological material. The system of a fully heated nozzle and a cooled print bed were developed to maintain the optimal hydrogel temperature and crosslinking speed. Our modifications of the bioprinter design improved the mechanical properties of the printouts and their accuracy while maintaining the maximal survival rate of cells and increasing the capacity of the bioink reservoir.

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 2 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.