Speaker
Description
Introduction
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. Some promising approaches include co-culture techniques, but traditional methods have drawbacks, including nutrient competition and instability due to different growth rates among the strains. Alternative strategies that retain the benefits of co-culture while avoiding its limitations are required. One such alternative comes from the field of biofabrication, which allows the creation of complex biological constructs in a high-throughput and cost-effective manner through encapsulation methods. While encapsulation and co-culture can be combined, most approaches create unequal volumes and interface areas, potentially leading to imbalanced growth. Is not an easy task to develop an encapsulation method able to maintain a co-culture that supports the cooperative growth of bacteria while maintaining stability over time.
Methodology
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 (Figure 1). The spatial arrangement of the multilayered architecture was designed to promote cooperative interactions (Figure 1b), particularly by embedding strict anaerobes between facultative anaerobes to create self-sustaining hypoxic niches. The printed constructs were characterized over 72 hours using fluorescence microscopy, colony-forming unit counts, LIVE/DEAD assays, and qPCR.
Results and discussion
Results showed that structured co-cultures exhibited higher viability, enhanced growth, and more balanced population dynamics than the monocultures of each bacterial strain and unstructured (scrambled) co-cultures.This study demonstrates that chaotic bioprinting enables precise spatial control over microbial ecosystems, allowing the rational design of microbial communities with tailored interactions. The approach presents a powerful and scalable platform for next-generation probiotic production and opens new opportunities for engineered microbiomes, synthetic biology, and living material design.
Keywords: biofabrication, probiotics, Next Generation Probiotics, chaotic printing, co-culture.
64057829897