Abstract
Three-dimensional (3D) printing is a transformative manufacturing strategy, allowing rapid prototyping, customization, and flexible manipulation of structure-property relationships. Proteins are particularly appealing to formulate inks for 3D printing as they serve as essential structural components of living systems, provide a support presence in and around cells and for tissue functions, and also provide the basis for many essential ex vivo secreted structures in nature. Protein-based inks are beneficial in vivo due to their mechanics, chemical and physical match to the specific tissue, and full degradability, while also to promoting implant-host integration and serving as an interface between technology and biology. Exploiting the biological, chemical, and physical features of protein-based inks can provide key opportunities to meet the needs of tissue engineering and regenerative medicine. Despite these benefits, protein-based inks impose nontrivial challenges to 3D printing such as concentration and rheological features and reconstitution of the structural hierarchy observed in nature that is a source of the robust mechanics and functions of these materials. This review introduces photo-crosslinking mechanisms and rheological principles that underpins a variety of 3D printing techniques. The review also highlights recent advances in the design, development, and biomedical utility of monolithic and composite inks from a range of proteins, including collagen, silk, fibrinogen, and others. One particular focus throughout the review is to introduce unique material characteristics of proteins, including amino acid sequences, molecular assembly, and secondary conformations, which are useful for designing printing inks and for controlling the printed structures. Future perspectives of 3D printing with protein-based inks are also provided to support the promising spectrum of biomedical research accessible to these materials.