Guiding Bacterial Activity for Biofabrication of Complex Materials via Controlled Wetting of Superhydrophobic Surfaces

Superhydrophobic surfaces are promising for preventing fouling and the formation of biofilms, with important implications in the food chain, maritime transport, and health sciences, among others. In this work, we exploit the interplay between wetting principles of superhydrophobic surfaces and microbial fouling for advanced three-dimensional (3D) biofabrication of biofilms. We utilize hydrostatic and capillary pressures to finely control the air-water interface and the aerotaxis-driven biofabrication on superhydrophobic surfaces. Superhydrophobic 3D molds are produced by a simple surface modification that partially embeds hydrophobic particles in silicone rubber. Thereafter, the molds allow the templating of the air-water interface of the culture medium, where the aerobic nanocellulose-producing bacteria (Komagataeibacter medellinensis) are incubated. The biofabricated replicas are hollow and seamless nanofibrous objects with a controlled morphology. Gradients of thickness, topographical feature size, and fiber orientation on the biofilm are obtained by controlling wetting, incubation time, and nutrient availability. Furthermore, we demonstrate that capillary length limitations are overcome by using pressurized closed molds, whereby a persistent air plastron allows the formation of 3D microstructures, regardless of their morphological complexity. We also demonstrate that interfacial biofabrication is maintained for at least 12 days without observable fouling of the mold surface. In summary, we achieve controlled biofouling of the air-water interface as imposed by the experimental framework under controlled wetting. The latter is central to both microorganism-based biofabrication and fouling, which are major factors connecting nanoscience, synthetic biology, and microbiology.