CAREER: Understanding Spatial Heterogeneity in Biofilms Using Colloidal Engineering

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Non-Technical Abstract:Many microorganisms, including bacteria and algae, form thin films on surfaces known as biofilms. Examples of these include dental plaque, the slime that forms on ship hulls, and wound-associated infections. Biofilms contain millions of microscopic cells, and the individual locations of these cells affect their ability to function. For example, the location of cells within a wound-associated biofilm affects their ability to resist antibiotics. To understand how cell location affects function, their locations must be controlled in three dimensions; however, because the microorganisms within a biofilm are microscopically small, they are difficult to manipulate at a single cell level. This CAREER award by the Biomaterials program in the Division of Materials Research to Montana State University, Bozeman, supports the investigation of biofilm structure and function by building biofilms from microscopic modules that can be assembled, disassembled, and analyzed in a controlled manner. This approach will enable researchers to synthetically construct multi-species biofilms in the laboratory to understand how bugs communicate, cooperate, or compete. The proposed work will benefit society as a whole by uncovering the physical mechanisms behind microbial virulence and antibiotic resistance. This understanding can lead to new methods that will aid in controlling these persistent, surface-associated microbial colonies. Impact in the state of Montana will be provided by a "Soft Materials Shop" (SMASH), which will be used for both outreach and teaching, and will strengthen the connection of sciences, engineering, architecture, and art on campus. This space will be the first in the state of Montana and will be accessible to the public. This proposal will help prepare Montana students for future careers in STEM using inquiry-based learning strategies and hands-on demos, educate the public about microfabrication and engineering, and provide opportunities for outreach that will engage public audiences from youth to adults. Technical AbstractBiofilms are ubiquitous in the natural world, are formed by nearly all microorganisms, including algae, bacteria, archaea, and can be both beneficial and harmful. The spatial locations of microbes in a biofilm impact biofilm function and physiology; however, methods for structuring biofilms in three dimensions are currently lacking. The ability to synthetically construct a heterogeneous, multi-species biofilm by placing microorganisms in discrete locations within a cultivated environment would allow researchers to bridge this gap and explore how bugs communicate, cooperate, or compete. To achieve this, colloidal engineering techniques will be applied to place microbes in spatially organized three-dimensional locations to understand how spatial heterogeneity influences structure-function relationships in biofilms. The aims of this proposal are to create monodisperse capsules of gels and shells to encapsulate microbes, to assemble capsules into three-dimensional structures, and finally to create modular synthetic consortia of bacteria to explore questions related to heterogeneity in biofilms. These constructs will be used to understand how biofilm behavior such as quorum sensing, matrix formation, and dormancy arises from heterogeneity in biofilms. This CAREER award by the Biomaterials program in the Division of Materials Research to Montana State University, Bozeman, will support the integration of soft matter science into classes, workshops, demonstrations, and tours within a dedicated space called "SMASH", the creation of K-8 after-school outreach modules, and an undergraduate/graduate course on "Soft Matter Engineering". This award will allow for creative, multidisciplinary collaboration, bringing together students in art, architecture, engineering, and science on campus to create, prototype, design, and build. The ideas, workshops, and demonstrations created in this space will be incorporated into current K-8 outreach efforts and extended to future outreach efforts to introduce soft matter science, a relatively young area of research, to students and the general public.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.