Bioprinting microbial harmony: Engineering spatially organized probiotic ecosystems via chaotic bioprinting Academic Article in Scopus uri icon

abstract

  • Probiotic therapies offer great potential for addressing gut dysbiosis, but current approaches are limited by low strain diversity, high production costs, and the challenges of culturing strict anaerobes. To overcome these limitations, this work introduces a novel strategy based on continuous chaotic bioprinting to create structured cocultures of probiotic bacteria. Using a Kenics static mixer¿based printhead, we fabricated alginate hydrogel filaments with an internal multilayered microarchitecture containing four probiotic strains: Bifidobacterium bifidum , Bacteroides fragilis , Lactobacillus rhamnosus , and Streptococcus thermophilus . The spatial arrangement of the multilayered architecture was designed to promote cooperative interactions, particularly by embedding strict anaerobes between facultative anaerobes to create self-sustaining hypoxic niches. The printed constructs were characterized over 72 h using fluorescence microscopy, colony-forming unit counts, LIVE/DEAD assays, qPCR, gas chromatography, and dynamic mechanic analysis. Results showed that structured cocultures exhibited higher viability, enhanced growth, and more balanced population dynamics than the monocultures of each bacterial strain and unstructured (scrambled) cocultures. Short-Chain Fatty Acid Production suggests functional cross-feeding; coculture filaments exhibited notably higher levels of acetic and propionic acids than the corresponding monocultures. This study demonstrates that chaotic bioprinting enables precise spatial control over microbial ecosystems, allowing the rational design of microbial communities with tailored interactions. Furthermore, chaotically printed constructs preserved probiotic viability for at least six weeks under storage, survived food-buffered gastric simulations, and maintained structural integrity over time, underscoring their robustness and translational potential. Chaotic bacterial bioprinting presents a powerful and scalable platform for next-generation probiotic production and opens new opportunities for engineered microbiomes, synthetic biology, and living material design. © 2025 Elsevier B.V.

publication date

  • December 1, 2025