Transparent Intracellular Sensing Platform with Si Needles for Simultaneous Live Imaging.

Vertically ordered Si needles are of particular interest for long-term intracellular recording owing to their capacity to infiltrate living cells with negligible damage and minimal toxicity. Such intracellular recordings could greatly benefit from simultaneous live cell imaging without disrupting their culture, contributing to an in-depth understanding of cellular function and activity. However, the use of standard live imaging techniques, such as inverted and confocal microscopy, is currently impeded by the opacity of Si wafers, typically employed for fabricating vertical Si needles. Here, we introduce a transparent intracellular sensing platform that combines vertical Si needles with a percolated network of Au-Ag nanowires on a transparent elastomeric substrate. This sensing platform meets all prerequisites for simultaneous intracellular recording and imaging, including electrochemical impedance, optical transparency, mechanical compliance, and cell viability. Proof-of-concept demonstrations of this sensing platform include monitoring electrical potentials in cardiomyocyte cells and in three-dimensionally engineered cardiovascular tissue, all while conducting live imaging with inverted and confocal microscopes. This sensing platform holds wide-ranging potential applications for intracellular research across various disciplines such as neuroscience, cardiology, muscle physiology, and drug screening.

Biodegradable silicon nanoneedles for ocular drug delivery.

Ocular drug delivery remains a grand challenge due to the complex structure of the eye. Here, we introduce a unique platform of ocular drug delivery through the integration of silicon nanoneedles with a tear-soluble contact lens. The silicon nanoneedles can penetrate into the cornea in a minimally invasive manner and then undergo gradual degradation over the course of months, enabling painless and long-term sustained delivery of ocular drugs. The tear-soluble contact lens can fit a variety of corneal sizes and then quickly dissolve in tear fluid within a minute, enabling an initial burst release of anti-inflammatory drugs. We demonstrated the utility of this platform in effectively treating a chronic ocular disease, such as corneal neovascularization, in a rabbit model without showing a notable side effect over current standard therapies. This platform could also be useful in treating other chronic ocular diseases.

Rapid custom prototyping of soft poroelastic biosensor for simultaneous epicardial recording and imaging.

The growing need for the implementation of stretchable biosensors in the body has driven rapid prototyping schemes through the direct ink writing of multidimensional functional architectures. Recent approaches employ biocompatible inks that are dispensable through an automated nozzle injection system. However, their application in medical practices remains challenged in reliable recording due to their viscoelastic nature that yields mechanical and electrical hysteresis under periodic large strains. Herein, we report sponge-like poroelastic silicone composites adaptable for high-precision direct writing of custom-designed stretchable biosensors, which are soft and insensitive to strains. Their unique structural properties yield a robust coupling to living tissues, enabling high-fidelity recording of spatiotemporal electrophysiological activity and real-time ultrasound imaging for visual feedback. In vivo evaluations of custom-fit biosensors in a murine acute myocardial infarction model demonstrate a potential clinical utility in the simultaneous intraoperative recording and imaging on the epicardium, which may guide definitive surgical treatments.

Acetogenic bacteria utilize light-driven electrons as an energy source for autotrophic growth.

Acetogenic bacteria use cellular redox energy to convert CO2 to acetate using the Wood-Ljungdahl (WL) pathway. Such redox energy can be derived from electrons generated from H2 as well as from inorganic materials, such as photoresponsive semiconductors. We have developed a nanoparticle-microbe hybrid system in which chemically synthesized cadmium sulfide nanoparticles (CdS-NPs) are displayed on the cell surface of the industrial acetogen Clostridium autoethanogenum The hybrid system converts CO2 into acetate without the need for additional energy sources, such as H2, and uses only light-induced electrons from CdS-NPs. To elucidate the underlying mechanism by which C. autoethanogenum uses electrons generated from external energy sources to reduce CO2, we performed transcriptional analysis. Our results indicate that genes encoding the metal ion or flavin-binding proteins were highly up-regulated under CdS-driven autotrophic conditions along with the activation of genes associated with the WL pathway and energy conservation system. Furthermore, the addition of these cofactors increased the CO2 fixation rate under light-exposure conditions. Our results demonstrate the potential to improve the efficiency of artificial photosynthesis systems based on acetogenic bacteria integrated with photoresponsive nanoparticles.

3D Printed Bioresponsive Devices with Selective Permeability Inspired by Eggshell Membrane for Effective Biochemical Conversion.

Eggshell membrane has selective permeability that enables gas or liquid molecules to pass through while effectively preventing migration of microbial species. Herein, inspired by the architecture of the eggshell membrane, we employ three-dimensional (3D) printing techniques to realize bioresponsive devices with excellent selective permeability for effective biochemical conversion. The fabricated devices show 3D conductive carbon nanofiber membranes in which precultured microbial cells are controllably deployed. The resulting outcome provides excellent selective permeability between chemical and biological species, which enables acquisition of target responses generated by biological species confined within the device upon input signals. In addition, electrically conductive carbon nanofiber networks provide a platform for real-time monitoring of metabolism of microbial cells in the device. The suggested platform represents an effort to broaden microbial applications by constructing biologically programmed devices for desired responses enabled by designated deployment of engineered cells in a securely confined manner within enclosed membranes using 3D printing methods.

Bactericidal Lubricating Synthetic Materials for Three-Dimensional Additive Assembly with Controlled Mechanical Properties.

3D printable synthetic materials have been developed to realize desired surface and mechanical properties. Lubricating synthetic surfaces have broad technological impacts on many applications including food packaging, microfluidic systems, and biomedical devices. However, combining soft materials with lubricants leads to significant phase separation and swelling phenomena, together with lowered mechanical strength, impeding full utilization of lubricating synthetic surfaces with desired shapes in a highly controllable manner. Here, we report a new platform to create a 3D printable lubricant-polymer composite (3D-LUBRIC) for the seamless fabrication of multidimensional structures with diverse functionalities. The rationally designed lubricant-polymer mixtures including silica aerogel particles not only exhibit suitable rheological properties for direct ink writing without phase separation but also enable the deterministic additive assembly of heterogeneous materials, which have large mismatches of oil permeability, with no distinct shape distortion. While exhibiting excellent lubricating properties for a variety of liquids, 3D-LUBRIC shows tunable mechanical properties with desired functionalities, such as optical transparency, flexibility and stretchability, and anti-icing and antibacterial/bactericidal properties. We employ the proposed platform to fabricate self-cleanable containers and antibacterial/bactericidal medical tubes. Our platform can offer new opportunities for building low-adhesive, multifunctional synthetic materials with customized shapes for diverse applications.

Bioresorbable, Miniaturized Porous Silicon Needles on a Flexible Water-Soluble Backing for Unobtrusive, Sustained Delivery of Chemotherapy.

Conventional melanoma therapies suffer from the toxicity and side effects of repeated treatments due to the aggressive and recurrent nature of melanoma cells. Less-invasive topical chemotherapies by utilizing polymeric microneedles have emerged as an alternative, but the sustained, long-lasting release of drug cargos remains challenging. In addition, the size of the microneedles is relatively bulky for the small, curvilinear, and exceptionally sensitive cornea for the treatment of ocular melanoma. Here, we report a design of bioresorbable, miniaturized porous-silicon (p-Si) needles with covalently linked drug cargos at doses comparable to those of conventional polymeric microneedles. The p-Si needles are built on a water-soluble film as a temporary flexible holder that can be intimately interfaced with the irregular surface of living tissues, followed by complete dissolution with saline solution within 1 min. Consequently, the p-Si needles remain embedded inside tissues and then undergo gradual degradation, allowing for sustained release of the drug cargos. Its utility in unobtrusive topical delivery of chemotherapy with minimal side effects is demonstrated in a murine melanoma model.

Hierarchical Macroporous Particles for Efficient Whole-Cell Immobilization: Application in Bioconversion of Greenhouse Gases to Methanol.

A viable approach for methanol production under ambient physiological conditions is to use greenhouse gases, methane (CH4) and carbon dioxide (CO2), as feed for immobilized methanotrophs. In the present study, unique macroporous carbon particles with pore sizes in the range of ∼1-6 µm were synthesized and used as support for the immobilization of Methylocella tundrae. Immobilization was accomplished covalently on hierarchical macroporous carbon particles. Maximal cell loading of covalently immobilized M. tundrae was 205 mgDCM g-1 of particles. Among these particles, the cells immobilized on 3.6 µm pore size particles showed the highest reusability with the least leaching and were chosen for further study. After immobilization, M. tundrae showed up to 2.4-fold higher methanol production stability at various pH and temperature values because of higher stability and metabolic activity than free cells. After eight cycles of reuse, the immobilized cells retained 18.1-fold higher relative production stability compared to free cells. Free and immobilized cells exhibited cumulative methanol production of 5.2 and 9.5 µmol mgDCM-1 under repeated batch conditions using simulated biogas [CH4 and CO2, 4:1 (v/v)] as feed, respectively. The appropriate pore size of macroporous particles favors the efficient M. tundrae immobilization to retain better biocatalytic properties. This is the first report concerning the covalent immobilization of methanotrophs on the newly synthesized macroporous carbon particles and its subsequent application in repeated methanol production using simulated biogas as a feed.