Citocompatibility Assessment of 3D-Printed Collagen Scaffolds with Hyaluronic Acid Crosslinked with Genipin, Dehydrothermal Treatment, and 1-Ethyl-3-(3-Dimethylaminopropyl) Carbodiimide and N-Hydroxysuccinimide

This study aimed to produce and evaluate the biocompatibility of 3D-printed collagen and hyaluronic acid scaffolds for soft tissue regeneration. Three different crosslinking methods were tested on the constructs. The first method is a chemical crosslinking with l-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS). The second method, dehydrothermal (DHT) treatment, is a physical approach that applies heat and pressure to induce crosslinking. Lastly, crosslinking with genipin, a natural agent, was also evaluated. Solubility tests in PBS at 37°C revealed optimal conditions for each crosslinking method, minimizing mass loss. SEM analysis demonstrated excellent cell adhesion and proliferation for Vero and BJ cell lines. Live/dead cell assays confirmed biocompatibility, showing high cell viability in contact with the scaffold extracts. These findings support the potential use of crosslinked collagen and HA scaffolds for soft tissue regeneration, providing a solid foundation for future research and clinical applications in tissue engineering.