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Bioelectrochemical technologies have attracted significant scientific interest because the effective bacterial electron exchange with external electrodes can provide a sustainable solution that joins environmental remediation and energy recovery. Multispecies electroactive bacterial biofilms are catalysts that will drive the operation of bioelectrochemical devices. Unfortunately, there is a lack of understanding of key mechanisms determining their electron-generating capabilities and syntrophic relations within microbial communities in biofilms. This is because there are no universally standardized models for simple, rapid, reliable, and cost-effective fabrication and characterization of electroactive multispecies biofilms. The heterogeneous and long-term nature of biofilm formation has hampered the development of those models. This work develops novel biofabrication and analysis platforms by creating innovative, paper-based 3-D systems that accurately recapitulate the structure, function, and physiology of living multispecies biofilms. Multiple layers of paper containing bacterial cells were stacked to simulate different layered 3-D biofilm models with defined cellular compositions and microenvironments. Overall bacterial electrogenic capabilities through the biofilm structures were characterized by thoroughly monitoring collective electron flows through different external resistors. Changes in the type of species and order of stacking created biofilm modeling which allowed for the study of their electrogenic performance via variation in electron flow rate output. Furthermore, multi-laminate structures allowed for straightforward de-stacking and layer-by-layer separation for analyses of pH distribution and cellular viability. Our multi-laminate structures provide a new strategy for (i) controlling the biofilm geometry of 3-D bacterial cultures, (ii) monitoring the microbial electoral properties, and (iii) constructing an artificial biofilm layer by layer. 

Traditional hydrophilic wound dressings, while common, fail to effectively drain wound exudate, creating conditions favorable for bacterial growth. Similarly, newer Janus‐type dressings with hydrophobic‐hydrophilic properties also fall short, as their hydrophobic side causes excessive dryness by pulling biofluids from the wound, disrupting moisture balance. Additionally, embedding antibiotics in dressings at fixed concentrations, regardless of the infection type, reduces effectiveness and contributes to the growing problem of antibiotic resistance. In response, a single‐layered Janus paper wound dressing, designed for efficient exudate absorption and precise antibiotic delivery, is developed. The approach differs from traditional Janus‐type dressings; a hydrophilic layer is placed directly against the wound for better moisture management, while antibiotics are applied through the hydrophobic layer. To further enhance exudate management, the hydrophilic section with four extra absorbent pads is extended. The dressing's antibiotic efficacy and dosage are tailored based on antibiotic susceptibility testing, ensuring targeted treatment. The selected antibiotic is manually added but automatically delivered directly to the wound bed. The in vitro and ex vivo evaluations, using bacterial cultures on agar and porcine skin assays, respectively, confirm the dressing's superior exudate drainage and its ability to inhibit pathogen growth and reproduction, marking a significant advancement in wound care.

 

An intimate and direct interface between inorganic electronics and living organisms will revolutionize the next generation of bioelectronics by bridging the signal and material gap between these two different fields. In this work, a redox-active microbial electrode is constructed as the novel interface by simultaneously 3-D printing and electropolymerizing 3,4-ethylenedioxythiophene (EDOT) in a liquid containing electrochemically active bacteria. A custom-made 3-D printer with a concurrent electrochemical control allows a scalable, template-free deposition of electrochemically active organic electrodes in a single printing. Electropolymerized poly(3,4-ethylenedioxythiophene) (PEDOT) acts as redox-active bridges by exploiting extracellularly transferred electrons generated from the bacterial respiration, constructing a seamless contact between the biological processes and the external abiotic systems. 

This work presents fabrication techniques for achieving individual electronic components both on the surface and within the fibers of a paper substrate, attaining full integration of paper and functional electronics materials. A process of hydrophobic wax patterning coupled with conductive and semiconductive poly(3,4-ethylenedioxythiophene): poly(styrene sulfonic acid) (PEDOT: PSS)-based ink injection and screen-printing has allowed for the implementation of all-paper-based, tunable resistors, capacitors, and transistors. The characteristics of the paper resistors can be adjusted as desired through finetuning of the PEDOT: PSS- based ink recipe, and the components can be combined in various arrangements to attain paper-based printed circuit boards (PCBs) for a wide range of practical applications. As a first step towards multiple component integration, a simple example circuit design is demonstrated that incorporates the three different components. Furthermore, through the strategic organization of the resistors, transistors, and capacitors and stacking of paper layers, more complex and diverse paper PCBs can be attained while minimizing the perceived surface area of the circuitry, allowing for a compact, pliable, and highly customizable means of fabricating paper-based electronic systems. 

Paper, an inexpensive material with natural biocompatibility, non‐toxicity, and biodegradability, allows for affordable and cost‐effective substrates for unconventional advanced electronics, often called papertronics. On the other hand, polymeric elastomers have shown to be an excellent success for substrates of soft bioelectronics, providing stretchability in skin wearable technology for continuous sensing applications. Although both materials hold their unique advantageous characteristics, merging both material properties into a single electronic substrate reimagines paper‐based bioelectronics for wearable and patchable applications in biosensing, energy generation and storage, soft actuators, and more. Here, a breathable, light‐weighted, biocompatible engineered stretchable paper is reported via coaxial nonwoven microfibers for unconventional bioelectronic substrates. The stretchable papers allow intimate bioconformability without adhesive through coaxial electrospinning of a cellulose acetate polymer (sheath) and a silicone elastomer (core). The fabricated cellulose‐silicone fibers exhibit a greater percent strain than commercially available paper while retaining hydrophilicity, biocompatibility, combustibility, disposable, and other natural characteristics of paper. Moreover, the nonwoven stretchable cellulose‐silicone fibrous mat can adapt conventional printing and fabrication process for paper‐based electronics, an essential aspect of advanced bioelectronic manufacturing.

 

Bacteria‐powered biobatteries using multiple microbial species under well‐mixed conditions demonstrate a temporary performance enhancement through their cooperative interaction, where one species produces a resource that another species needs but cannot synthesize. Despite excitement about the artificial microbial consortium, those mixed populations cannot be robust to environmental changes and have difficulty generating long‐lasting power because individual species compete with their neighbors for space and resources. In nature, microbial communities are organized spatially as multiple species are separated by a few hundred micrometers to balance their interaction and competition. However, it has been challenging to define a microscale spatial microbial structure in miniature biobatteries. Here, an innovative technique to design microscale spatial structures with microbial multispecies for significant improvement of the biobattery performance is demonstrated. A solid‐state layer‐by‐layer agar‐based culture platform is proposed, where individual microcolonies separately confined in microscale agar layers form a 3‐D spatial structure allowing for the exchange of metabolites without physical contact between the individual species. The optimized microbial co‐cultures are determined from selected hypothesis‐driven naturally‐occurring bacteria. Vertically and horizontally structured 3‐D microbial communities in solid‐state agar‐based microcompartments demonstrate the practicability of the biobattery, generating longer and greater power in a more self‐sustaining manner than monocultures and other mixed populations.