Using chaotic printing to fabricate environments that support the culture of requisitive bacteria strains and bacterial communities

Abstract

This thesis explores the use of 3D bioprinting technologies in enhancing the efficacy and resilience of different bacterial strains contained within the human gut microbiota. By encapsulating these different bacterial species in hydrogel fibers, the study aims to create tailored bacterial mono and co-cultures that overcome the limitations of standard probiotic formulation and delivery, such as susceptibility to environmental stressors and poor colonization capacities while also addressing challenges in the generation and culture of bacterial consortia such as the adequate proliferation of all the members in the community. Our research demonstrates that chaotic bioprinting offers a novel platform for the high-throughput culture and production of probiotics. We successfully cultivated Bifidobacterium bifidum in bioprinted alginate fibers, achieving a viability of 1x1012 CFU/g of fiber within the first 8 hours post-printing, without incubation under a strict anaerobic atmosphere. This highlights the potential of 3D bioprinting in maintaining probiotic viability and enhancing their therapeutic benefits. Furthermore, the study delves into the spatial and growth dynamics of mixed and spatially arranged bacterial communities. By examining communities comprising Escherichia coli, Lactobacillus rhamnosus, and B. bifidum, we demonstrate how chaotic advection influences the arrangement and development of these species. Our findings reveal that the spatial arrangement of bacterial species plays a crucial role in their resilience and development within the community, offering insights into optimizing probiotic formulations and in vitro study models for improved human health benefits. This work contributes to the blooming field of microbiome science; our research paves the way for more effective and resilient probiotic treatments and better understanding the interactions occurring within the microbial world.