Enabling 3D bioprinting of cell-laden pure collagen scaffolds via tannic acid supporting bath.

The fabrication of cell-laden biomimetic scaffolds represents a pillar of tissue engineering and regenerative medicine (TERM) strategies, and collagen is the gold standard matrix for cells to be. In the recent years, extrusion 3D bioprinting introduced new possibilities to increase collagen scaffold performances thanks to the precision, reproducibility, and spatial control. However, the design of pure collagen bioinks represents a challenge, due to the low storage modulus and the long gelation time, which strongly impede the extrusion of a collagen filament and the retention of the desired shape post-printing. In this study, the tannic acid-mediated crosslinking of the outer layer of collagen is proposed as strategy to enable collagen filament extrusion. For this purpose, a tannic acid solution has been used as supporting bath to act exclusively as external crosslinker during the printing process, while allowing the pH- and temperature-driven formation of collagen fibers within the core. Collagen hydrogels (concentration 2-6 mg/mL) were extruded in tannic acid solutions (concentration 5-20 mg/mL). Results proved that external interaction of collagen with tannic acid during 3D printing enables filament extrusion without affecting the bulk properties of the scaffold. The temporary collagen-tannic acid interaction resulted in the formation of a membrane-like external layer that protected the core, where collagen could freely arrange in fibers. The precision of the printed shapes was affected by both tannic acid concentration and needle diameter and can thus be tuned. Altogether, results shown in this study proved that tannic acid bath enables collagen bioprinting, preserves collagen morphology, and allows the manufacture of a cell-laden pure collagen scaffold.

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The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision.