Cell-Laden Biomimetically Mineralized Shark-Skin-Collagen-Based 3D Printed Hydrogels for the Engineering of Hard Tissues.

Mineralization processes based on coprecipitation methods have been applied as a promising alternative to the most commonly used methods of polymer-ceramic combination, direct mixing, and incubation in simulated body fluid (SBF) or modified SBF. In the present study, for the first time, the mineralization (ideally hydroxyapatite formation) of blue shark ( (PG)) collagen to fabricate 3D printable cell-laden hydrogels is proposed. In the first part, several parameters for collagen mineralization were tested until optimization. The hydroxyapatite formation was confirmed by FT-IR, XRD, and TEM techniques. In the second part, stable bioinks combining the biomimetically mineralized collagen with alginate (AG) (1:1, 1:2, 1:3, and AG) solution were used for 3D printing of hydrogels. The addition of Ca ions into the system did present a synergistic effect: by one side, the mineralization of the collagen occurred, and at same time, they were also useful to ionically cross-link the blends with alginate, avoiding the addition of any cytotoxic chemical cross-linking agent. Mouse fibroblast cell line survival during and after printing was favored by the presence of PG collagen as exhibited by the biological performance of the hydrogels. Inspired in a concept of marine byproduct valorization, 3D bioprinting of mineralized blue shark collagen is thus proposed as a promising approach, envisioning the engineering of mineralized tissues.