<scp>3D</scp>‐Printed Gelatin‐Based Scaffold Crosslinked by Genipin: Evaluation of Mechanical Properties and Biological Effect

In this study, scaffolds based on natural polymer gelatin A, blended with polyvinylpyrrolidone were crosslinked by genipin (0.5 and 1 wt%), in order to investigate their mechanical performance and potential for biomedical application. Semi-solid extrusion (SSE) 3D printing technique was used, enabling in situ crosslinking of the blend during processing. Swelling test showed that the swelling ratio reduces with higher concentration of genipin due to an increased crosslinking. The FTIR analysis confirmed the crosslinking of scaffolds by genipin. DSC analysis and mechanical testing have shown improved thermal and mechanical properties. Morphological analysis of scaffolds by FESEM showed increased toughening of the material with the crosslinking. Tensile strength and microhardness showed a significant rise in scaffolds with the increase in genipin content, which was up to 93.8% and 125.3%, respectively. These findings were in accordance with morphological features present in samples. The biological effect of the scaffold matrix system was evaluated by qualitative and quantitative cytotoxicity assessment in vitro, demonstrating the absence of cytotoxicity in tested preparations in a direct test. The cytotoxicity index based on the metabolic activity of cells in an indirect test showed up to 20% reduction of viability compared with the control, confirming the absence of cytotoxicity, which was additionally verified by propidium iodine staining of the cells exposed to scaffolds. The presented gelatin-based crosslinked scaffolds obtained by 3D printing represent good candidates for biomedical application and future research that includes further in vitro and in vivo analysis.