Natural polymer derived hydrogel bioink with enhanced thixotropy improves printability and cellular preservation in 3D bioprinting

Three-dimensional (3D) bioprinting is evolving into a promising technology by spatially controlling the distribution of living cells for the biomedical field. However, maintaining high printability while protecting cells from damage due to shear stress remains the key challenge for extrusion-based 3D bioprinting. Herein, we developed a novel “protein-polyphenol-polysaccharide” extrusion-based bioink named Gel-TA-Alg@Ca²⁺ using gelatin (Gel), tannic acid (TA) and sodium alginate (Alg) with quantitative thixotropy by pre-crosslinking with a series of low concentrations of CaCl₂ at 0.03, 0.04, 0.05 and 0.06 M, respectively. Our experimental design quantitatively presented the positive proportional functional relationship between the thixotropy of Gel-TA-Alg@Ca²⁺ and printability (including injectability and formability) for the first time. Importantly, the thixotropy proportionately and significantly elevated cellular viability after 3D bioprinting due to the reduced extrusion force involved in printing. 3D bioprinted constructs composed of Gel-TA-Alg@Ca²⁺ and MG-63 cells exhibited a good cell viability rate for more than 14 days. These findings provide valuable insights into the rational design of thixotropic bioink and offer more opportunities to probe the relationship between the thixotropy and the success of 3D bioprinting.