Self-Supported Printing of Gelatin Composite-Based Engineered Living Materials

Abstract The emergence of engineered living materials (ELMs) has led to the development of functional composites by combining living microorganisms with nonliving components, particularly hydrogels. Hydrogels, which mimic the extracellular matrix, support microbial growth by providing essential nutrients and promoting cell adhesion, making them ideal for ELM production. However, hydrogel-based materials often face challenges in three-dimensional printing due to poor structural integrity and limited printability, frequently requiring additional processes, precise control, and/or material modifications to enhance their printing performance. This study focuses on developing a microorganism-laden gelatin microgel and gelatin solution-based composite bioink for self-supported printing of ELMs, enhanced via microbial-induced calcium carbonate precipitation. Gelatin microgels are utilized as rheology modifiers, enabling the yield-stress fluid behavior of the bioink for improved printability and postprinting shape retention, while transglutaminase enzymatically cross-links printed structures completely, resulting in good printability. Furthermore, Sporosarcina pasteurii in the bioink enables calcium carbonate deposition during postprinting culturing, forming robust, biomineralized structures. Fabricated samples are found to have significant successful mineral deposition with over 50 wt% calcium carbonate content, and they exhibit compressive strengths of up to 1.4 MPa. This approach offers a cost-effective, energy-efficient method for creating high-strength, biocompatible biocomposites with potential applications such as bone tissue engineering, coral restoration, and sustainable building development.