A microfluidic platform for the synthesis of polymer and polymer-protein-based protocells.

In this study, we demonstrate the fabrication of polymersomes, protein-blended polymersomes, and polymeric microcapsules using droplet microfluidics. Polymersomes with uniform, single bilayers and controlled diameters are assembled from water-in-oil-in-water double-emulsion droplets. This technique relies on adjusting the interfacial energies of the droplet to completely separate the polymer-stabilized inner core from the oil shell. Protein-blended polymersomes are prepared by dissolving protein in the inner and outer phases of polymer-stabilized droplets. Cell-sized polymeric microcapsules are assembled by size reduction in the inner core through osmosis followed by evaporation of the middle phase. All methods are developed and validated using the same glass-capillary microfluidic apparatus. This integrative approach not only demonstrates the versatility of our setup, but also holds significant promise for standardizing and customizing the production of polymer-based artificial cells.

Cellulose acetate microwell plates for high-throughput colorimetric assays.

Single use plasticware (SUP) in scientific, diagnostic, and academic laboratories makes a significant contribution to plastic waste generation worldwide. Polystyrene (PS) microwell plates form a part of this waste. These plates are the backbone of high throughput colorimetric measurements in academic, research, and healthcare settings for detection/quantification of wide-ranging analytes including proteins, carbohydrates, nucleic acids, and enzyme activity. Polystyrene (PS) microwell plates serve as a platform for holding samples and reagents, where mixing initiates chemical reaction(s), and the ensuing color changes are quantified using a microplate reader. However, these plates are rarely reused or recycled, contributing to the staggering amounts of plastic waste generated in scientific laboratories. Here, we are reporting the fabrication of cellulose acetate (CA) microwell plates as a greener alternative to non-biodegradable PS plates and we demonstrate their application in colorimetric assays. These easy to fabricate, lighter weight, customizable, and environmentally friendly plates were fabricated in 96- and 384-well formats and made water impermeable through chemical treatment. The plates were tested in three different colorimetric analyses: (i) bicinchoninic acid assay (BCA) for protein quantification; (ii) chymotrypsin (CT) activity assay; and (iii) alkaline phosphatase (AP) activity assay. Color intensities were quantified using a freely available smartphone application, Spotxel® Reader (Sicasys Software GmbH). To benchmark the performance of this platform, the same assays were performed in commercial PS plates too and quantified using a UV/Vis microplate reader. The two systems yielded comparable linear correlation coefficients, LOD and LOQ values, thereby validating the CA plate-cell phone based analytical method. The CA microwell plates, coupled with smart phone optical data capture, provide greener, accessible, and scalable tools for all laboratory settings and are particularly well-suited for resource- and infrastructure-limited environments.

SCO-pH: Microfluidic dynamic phenotyping platform for high-throughput screening of single cell acidification.

Studies on the dynamics of single cell phenotyping have been hampered by the lack of quantitative high-throughput metabolism assays. Extracellular acidification, a prominent phenotype, yields significant insights into cellular metabolism, including tumorigenicity. Here, we develop a versatile microfluidic system for single cell optical pH analysis (SCO-pH), which compartmentalizes single cells in 140-pL droplets and immobilizes approximately 40,000 droplets in a two-dimensional array for temporal extracellular pH analysis. SCO-pH distinguishes cells undergoing hyperglycolysis induced by oligomycin A from untreated cells by monitoring their extracellular acidification. To facilitate pH sensing in each droplet, we encapsulate a cell-impermeable pH probe whose fluorescence intensities are quantified. Using this approach, we can differentiate hyperglycolytic cells and concurrently observe single cell heterogeneity in extracellular acidification dynamics. This high-throughput system will be useful in applications that require dynamic phenotyping of single cells with significant heterogeneity.

Colloidal synthesis of metallodielectric Janus matchsticks.

We present a gram-scale synthesis of metallodielectric Janus matchsticks, which feature a gold-coated silica sphere and a silica rod. SiO2 Janus matchsticks are synthesized in one batch by growing amine-functionalized SiO2 spheres at the end of SiO2 rods. Gold deposition on the spheres produces Au-SiO2 Janus matchsticks with an aspect ratio controlled by the rod length. The metallodielectric Janus matchsticks, produced by scalable colloidal synthesis, hold great potential as functional colloidal materials.

Drug-loaded adhesive microparticles for biofilm prevention on oral surfaces.

The oral cavity, a warm and moist environment, is prone to the proliferation of microorganisms like Candida albicans (C. albicans), which forms robust biofilms on biotic and abiotic surfaces, leading to challenging infections. These biofilms are resistant to conventional treatments due to their resilience against antimicrobials and immune responses. The dynamic nature of the oral cavity, including the salivary flow and varying surface properties, complicates the delivery of therapeutic agents. To address these challenges, we introduce dendritic microparticles engineered for enhanced adhesion to dental surfaces and effective delivery of antifungal agents and antibiofilm enzymes. These microparticles are fabricated using a water-in-oil-in-water emulsion process involving a blend of poly(lactic-co-glycolic acid) (PLGA) random copolymer (RCP) and PLGA-b-poly(ethylene glycol) (PLGA-b-PEG) block copolymer (BCP), resulting in particles with surface dendrites that exhibit strong adhesion to oral surfaces. Our study demonstrates the potential of these adhesive microparticles for oral applications. The adhesion tests on various oral surfaces, including dental resin, hydroxyapatite, tooth enamel, and mucosal tissues, reveal superior adhesion of these microparticles compared to conventional spherical ones. Furthermore, the release kinetics of nystatin from these microparticles show a sustained release pattern that can kill C. albicans. The biodegradation of these microparticles on tooth surfaces and their efficacy in preventing fungal biofilms have also been demonstrated. Our findings highlight the effectiveness of adhesive microparticles in delivering therapeutic agents within the oral cavity, offering a promising approach to combat biofilm-associated infections.

Nonmonotonic polymer translocation kinetics through nanopores under changing surface-polymer interactions.

Understanding the dynamics of polymers in confined environments is pivotal for diverse applications ranging from polymer upcycling to bioseparations. In this study, we develop an entropic barrier model using self-consistent field theory that considers the effect of attractive surface interactions, solvation, and confinement on polymer kinetics. In this model, we consider the translocation of a polymer from one cavity into a second cavity through a single-segment-width nanopore. We find that, for a polymer in a good solvent (i.e., excluded volume, u0 > 0), there is a nonmonotonic dependence of mean translocation time (τ) on surface interaction strength, ɛ. At low ɛ, excluded volume interactions lead to an energetic penalty and longer translocation times. As ɛ increases, the surface interactions counteract the energetic penalty imposed by excluded volume and the polymer translocates faster through the nanopore. However, as ɛ continues to increase, an adsorption transition occurs, which leads to significantly slower kinetics due to the penalty of desorption from the first cavity. The ɛ at which this adsorption transition occurs is a function of the excluded volume, with higher u0 leading to an adsorption transition at higher ɛ. Finally, we consider the effect of translocation across different size cavities. We find that the kinetics for translocation into a smaller cavity speeds up while translocation to a larger cavity slows down with increasing ɛ due to higher surface contact under stronger confinement.

Flexible Antireflection Coatings with Enhanced Durability and Antifogging Properties.

Antireflection coatings (ARCs) enhance optical clarity and improve light transmission by reducing glare and reflections. The application of conventional ARCs in flexible devices, however, is impeded by their lack of durability, particularly under bending deformation. We develop ARCs that withstand delamination and fracture, remaining intact even after 1000 bending cycles with a 5 cm bending radius. We fabricate integrated ARCs (iARCs) on a poly(methyl methacrylate) (PMMA) substrate by inducing free polymers to infiltrate the interstices of a disordered assembly of hollow silica nanochains and nanospheres. The polydispersity of PMMA creates a refractive index gradient, yielding a broadband antireflection capability. The nanochain-based iARCs are superior to the nanosphere-based coatings in both antireflection properties and mechanical durability, owing to the lower packing density and mechanical interlocking of the nanochains, respectively. Additionally, these nanochain iARCs display antifogging properties stemming from their superhydrophilicity. While our demonstrations are based on PMMA as a model substrate, this methodology is potentially extendable to other polymers, enhancing the iARC's applicability across various practical applications, including flexible and wearable devices.

Increase in the effective viscosity of polyethylene under extreme nanoconfinement.

Understanding polymer transport in nanopores is crucial for optimizing heterogeneously catalyzed processes in polymer upcycling and fabricating high-performance nanocomposite films and membranes. Although confined polymer dynamics have been extensively studied, the behavior of polyethylene (PE)-the most widely used commodity polymer-in pores smaller than 20 nm remains largely unexplored. We investigate the effects of extreme nanoconfinement on PE transport using capillary rise infiltration in silica nanoparticle packings with average pore radii ranging from ∼1 to ∼9 nm. Using in situ ellipsometry and the Lucas-Washburn model, we discover a previously unknown inverse relationship between effective viscosity (ηeff) and average pore radius (Rpore). Additonally, we determine that PE transport under these extreme conditions is primarily governed by physical confinement, rather than pore surface chemistry. We refine an existing theory to provide a generalized formalism to describe the polymer transport dynamics over a wide range of pore radii (from 1 nm and larger). Our results offer valuable insights for optimizing catalyst supports in polymer upcycling and improving infiltration processes for nanocomposite fabrication.

Capillary filling dynamics of polymer melts in a bicontinuous nanoporous scaffold.

Polymer infiltrated nanoporous gold is prepared by infiltrating polymer melts into a bicontinuous, nanoporous gold (NPG) scaffold. Polystyrene (PS) films with molecular weights (Mw) from 424 to 1133 kDa are infiltrated into a NPG scaffold (∼120 nm), with a pore radius (Rp) and pore volume fraction of 37.5 nm and 50%, respectively. The confinement ratios (Γ=RgRp) range from 0.47 to 0.77, suggesting that the polymers inside the pores are moderately confined. The time for PS to achieve 80% infiltration (τ80%) is determined using in situ spectroscopic ellipsometry at 150 °C. The kinetics of infiltration scales weaker with Mw, τ80%∝Mw1.30±0.20, than expected from bulk viscosity Mw3.4. Furthermore, the effective viscosity of the PS melt inside NPG, inferred from the Lucas-Washburn model, is reduced by more than one order of magnitude compared to the bulk. Molecular dynamics simulation results are in good agreement with experiments predicting scaling as Mw1.4. The reduced dependence of Mw and the enhanced kinetics of infiltration are attributed to a reduction in chain entanglement density during infiltration and a reduction in polymer-wall friction with increasing polymer molecular weight. Compared to the traditional approach involving adding discrete particles into the polymer matrix, these studies show that nanocomposites with higher loading can be readily prepared, and that kinetics of infiltration are faster due to polymer confinement inside pores. These films have potential as actuators when filled with stimuli-responsive polymers as well as polymer electrolyte and fuel cell membranes.

Tension-activated nanofiber patches delivering an anti-inflammatory drug improve repair in a goat intervertebral disc herniation model.

Conventional microdiscectomy treatment for intervertebral disc herniation alleviates pain but does not repair the annulus fibrosus, resulting in a high incidence of recurrent herniation and persistent dysfunction. The lack of repair and the acute inflammation that arise after injury can further compromise the disc and result in disc-wide degeneration in the long term. To address this clinical need, we developed tension-activated repair patches (TARPs) for annulus fibrosus repair and local delivery of the anti-inflammatory factor anakinra (a recombinant interleukin-1 receptor antagonist). TARPs transmit physiologic strain to mechanically activated microcapsules embedded within the patch, which release encapsulated bioactive molecules in direct response to spinal loading. Mechanically activated microcapsules carrying anakinra were loaded into TARPs, and the effects of TARP-mediated annular repair and anakinra delivery were evaluated in a goat model of annular injury in the cervical spine. TARPs integrated with native tissue and provided structural reinforcement at the injury site that prevented aberrant disc-wide remodeling resulting from detensioning of the annular fibrosus. The delivery of anakinra by TARP implantation increased matrix deposition and retention at the injury site and improved maintenance of disc extracellular matrix. Anakinra delivery additionally attenuated the inflammatory response associated with TARP implantation, decreasing osteolysis in adjacent vertebrae and preserving disc cellularity and matrix organization throughout the annulus fibrosus. These results demonstrate the therapeutic potential of TARPs for the treatment of intervertebral disc herniation.

Bioinspired Switchable Passive Daytime Radiative Cooling Coatings.

Passive daytime radiative cooling (PDRC) relies on simultaneous reflection of sunlight and radiation toward cold outer space. Current designs of PDRC coatings have demonstrated potential as eco-friendly alternatives to traditional electrical air conditioning (AC). While many features of PDRC have been individually optimized in different studies, for practical impact, it is essential for a system to demonstrate excellence in all essential aspects, like the materials that nature has created. We propose a bioinspired PDRC structure templated by bicontinuous interfacially jammed emulsion gels (bijels) that possesses excellent cooling, thinness, tunability, scalability, and mechanical robustness. The unique bicontinuous disordered structure captures key features of Cyphochilus beetle scales, enabling a thin (130 µm) bijel PDRC coating to achieve high solar reflectance (≳0.97) and high longwave-infrared (LWIR) emissivity (≳0.93), resulting in a subambient temperature drop of ∼5.6 °C under direct sunlight. We further demonstrate switchable cooling inspired by the exoskeleton of the Hercules beetle and mechanical robustness in analogy to spongy bone structures.

Asymmetry-Enhanced Motion of Urease-Powered Micromotors from Double Emulsion-Templated Microcapsules.

Autonomous motion of enzyme-powered motors has important implications for drug delivery, cell-cell communication, and protocell engineering. Although many of these systems are inspired by the motion of biological cells, most of them lack key structural features, like micrometer-sized boundaries and aqueous compartments, and rely on bubble propulsion to generation motion. In this study, we use droplet microfluidics to generate large populations of cell-sized microcapsules with poly(lactic-co-glycolic acid) shells and functionalize their surfaces with the enzyme urease to drive their motion. We adjust the number of surface functional groups for urease conjugation by preparing microcapsules with two different surfactants, poly(vinyl alcohol) (PVA) and poly(ethylene-alt-maleic anhydride) (PEMA). We also tune the surface roughness of the microcapsules by varying the concentration of silica nanoparticles in the droplet middle phase. We find that PEMA plays a crucial role in increasing the grafting density of urease on the surface of smooth microcapsules, leading to active motion in the presence of urea. In addition, rough microcapsules prepared with PEMA and loaded with comparable amounts of urease move up to three times faster than their smooth counterparts, which we believe is due to an asymmetric distribution of urease on the surface, giving rise to a preferred direction of motion. Taken together, these results provide new insights into the role that various stabilizing agents play in the induction of motion by enzymatic motors prepared from microfluidics, which is a potentially powerful tool for future preparation of motile protocells in biomedicine.

Dual Porosity-Enhanced Antireflection Coatings with Continuous Gradient.

The incorporation of porous structures into films and coatings can transform their properties for applications in optics, separation, electronics, and energy generation and storage. Packing nanoparticles (NPs) is a versatile approach for fabricating nanoporous films with a tunable structure and properties. The mechanical fragility of NP packing-based films and coatings, however, significantly impedes their widespread utilization. Although infiltrating a polymer into the interstices of these NP packings has been shown to enhance their mechanical durability, this method completely eliminates the porosity of the structures, compromising their properties and functionality. This study presents a new approach to fabricate highly loaded porous nanocomposite films with a gradient in the refractive index by infiltrating subsaturating amounts of poly(methyl methacrylate) (PMMA) into disordered packings of hollow silica NPs. We demonstrate that dual porosity is a critical feature that enhances their antireflection (AR) and mechanical properties. The hollow cores of NPs prevent a substantial increase in the refractive index of the resulting films. Moreover, the interparticle voids allow for mechanical reinforcement to occur when the NP packings are infiltrated with PMMA, making them even more suitable for AR coatings. The refractive index and gradient across the nanocomposites can be tailored by adjusting the amount of PMMA infiltrated into the NP packing, the shape of hollow NPs, and the annealing time. The nanocomposite coatings with a continuous gradient in refractive index exhibit excellent AR properties and enhanced mechanical durability. Combined with the unique structural tunability afforded by the dual porosity, this approach provides a scalable and effective way to create robust and graded nanoporous structures for various applications.

Throughput-scalable manufacturing of SARS-CoV-2 mRNA lipid nanoparticle vaccines.

Lipid nanoparticles (LNPs) are a potent delivery technology that have made it possible for the recent clinical breakthroughs in mRNA therapeutics and vaccines. A key challenge to the broader implementation of mRNA therapeutics and vaccines is the development of technology to produce precisely defined LNP formulations, with throughput that can scale from discovery to commercial manufacturing and meet the stringent manufacturing standards of the pharmaceutical industry. To address these challenges, we have developed a microfluidic chip that incorporates 1×, 10×, or 256× LNP-generating units that achieve scalable production rates of up to 17 L/h of precisely defined LNPs. Using these chips, we demonstrate that LNP physical properties and potency in vivo are unchanged as throughput is scaled. Our chips are fabricated out of silicon and glass substrates, which have excellent solvent compatibility, compatibility with pharmaceutical manufacturing, and can be fully reset and reused. SARS-CoV-2 mRNA-LNP vaccines formulated by our chips triggered potent antibody responses in a preclinical study. These results demonstrate the feasibility of directly translating microfluidic-generated LNPs to the scale necessary for commercial production.

Synthesis of Triazole and Tetrazole-Functionalized Zr-Based Metal-Organic Frameworks Through Post-Synthetic Ligand Exchange.

Metal-organic frameworks (MOFs) are a class of porous materials that are formed through coordination bonds between metal clusters and organic ligands. Given their coordinative nature, the organic ligands and strut framework can be readily removed from the MOF and/or exchanged with other coordinative molecules. By introducing target ligands to MOF-containing solutions, functionalized MOFs can be obtained with new chemical tags via a process called post-synthetic ligand exchange (PSE). PSE is a straightforward and practical approach that enables the preparation of a wide range of MOFs with new chemical tags via a solid-solution equilibrium process. Furthermore, PSE can be performed at room temperature, allowing the incorporation of thermally unstable ligands into MOFs. In this work, we demonstrate the practicality of PSE by using heterocyclic triazole- and tetrazole-containing ligands to functionalize a Zr-based MOF (UiO-66; UiO = University of Oslo). After digestion, the functionalized MOFs are characterized via various techniques, including powder X-ray diffraction and nuclear magnetic resonance spectroscopy.

Water-Induced Separation of Polymers from High Nanoparticle-Content Nanocomposite Films.

Polymer nanocomposites with high loadings of nanoparticles (NPs) exhibit exceptional mechanical and transport properties. Separation of polymers and NPs from such nanocomposites is a critical step in enabling the recycling of these components and reducing the potential environmental hazards that can be caused by the accumulation of nanocomposite wastes in landfills. However, the separation typically requires the use of organic solvents or energy-intensive processes. Using polydimethylsiloxane (PDMS)-infiltrated SiO2 NP films, we demonstrate that the polymers can be separated from the SiO2 NP packings when these nanocomposites are exposed to high humidity and water. The findings indicate that the charge state of the NPs plays a significant role in the propensity of water to undergo capillary condensation within the PDMS-filled interstitial pores. We also show that the size of NPs has a crucial impact on the kinetics and extent of PDMS expulsion, illustrating the importance of capillary forces in inducing PDMS expulsion. We demonstrate that the separated polymer can be collected and reused to produce a new nanocomposite film. The work provides insightful guidelines on how to design and fabricate end-of-life recyclable high-performance nanocomposites.

Polymer-Grafted, Gold Nanoparticle-Based Nano-Capsules as Reversible Colorimetric Tensile Strain Sensors.

Colloidal colorimetric microsensors enable the in-situ detection of mechanical strains within materials. Enhancing the sensitivity of these sensors to small scale deformation while enabling reversibility of the sensing capability would expand their utility in applications including biosensing and chemical sensing. In this study, we introduce the synthesis of colloidal colorimetric nano-sensors using a simple and readily scalable fabrication method. Colloidal nano sensors are prepared by emulsion-templated assembly of polymer-grafted gold nanoparticles (AuNP). To direct the adsorption of AuNP to the oil-water interface of emulsion droplets, AuNP (≈11nm) are functionalized with thiol-terminated polystyrene (PS, Mn  = 11k). These PS-grafted gold nanoparticles are suspended in toluene and subsequently emulsified to form droplets with a diameter of ≈30µm. By evaporating the solvent of the oil-inwater emulsion, we form nanocapsules (AuNC) (diameter < 1µm) decorated by PS-grafted AuNP. To test mechanical sensing, the AuNC are embedded in an elastomer matrix. The addition of a plasticizer reduces the glass transition temperature of the PS brushes, and in turn imparts reversible deformability to the AuNC. The plasmonic peak of the AuNC shifts towards lower wavelengths upon application of uniaxial tensile tension, indicating increased inter-nanoparticle distance, and reverts back as the tension is released.

Modulation of Oil/Polymer Nanocapsule Size via Phase Diagram-Guided Microfluidic Coprecipitation.

Flow-based nanoprecipitation of different solutes via rapid mixing of two miscible liquids is a scalable strategy for manufacturing nanoparticles with various shapes and morphologies. Controlling the size of nanoparticles in flow-based nanoprecipitation, however, is often left to empirical variations in the flow rate ratios or the total flow rate of the two streams. In this work, we investigate the coprecipitations of oil and polymer to form nanocapsules via the Ouzo effect using glass capillary microfluidics across a range of mixing conditions. In the range of flow rates studied, the two streams mix convectively in micro-vortices formed at the junction of the two stream inlets. Using computational fluid dynamics simulations and glass capillary microfluidic nanoprecipitation, we establish a relationship between the precipitation conditions occurring experimentally in situ and the location on the ternary Ouzo phase diagram where precipitation is taking place. We find that a key variable in the resulting average diameter of the fabricated capsules is the degree of supersaturation experienced by both the oil and the polymer in the vortex zone of the device, showing a strong correlation between the two values. The control over the nanocapsule size by varying the extent of supersaturation of both precipitants is demonstrated by using two oils having distinct phase diagrams. This work provides a systematic approach to controlling the size of nanoparticles fabricated via continuous nanoprecipitation by linking the in situ flow conditions to ternary phase diagram behavior, enabling accurate control over nanocapsule size.

Wetting ridges on slippery liquid-infused porous surfaces.

Slippery liquid-infused porous surfaces (SLIPS) show remarkable liquid repellency, making them useful for many coating applications. The outstanding repellency of SLIPS comes from a lubricant layer stabilized within and at the surface of a porous template. The stability of this lubricant layer is key for SLIPS to exhibit their unique functionality. The lubricant layer, however, is depleted over time, causing degradation of liquid repellency. The formation of wetting ridges surrounding liquid droplets on the surface of SLIPS is one of the primary sources of lubricant depletion. Here, we present the fundamental understanding and characteristics of wetting ridges and highlight the latest developments that enable the detailed investigation and suppression of wetting ridge formation on SLIPS. In addition, we offer our perspectives on new and exciting directions for SLIPS.

Brain Targeted Xenon Protects Cerebral Vasculature After Traumatic Brain Injury.

Abstract Cerebrovascular dysfunction following traumatic brain injury (TBI) is a well-characterized phenomenon. Given the therapeutic potential of xenon, we aimed to study its effects after localized delivery to the brain using microbubbles. We designed xenon-containing microbubbles stabilized by dibehenoylphosphatidylcholine (DBPC) and polyethylene glycol (PEG) attached to saturated phospholipid (DPSE-PEG5000). Using a pig model of TBI, these microbubbles were intravenously injected, and ultrasound was used to release xenon at the level of the carotid artery. The control group received perfluorobutane containing microbubbles. Diffusion tensor imaging (DTI) showed areas of higher fractional anisotropy for pigs receiving xenon microbubbles compared to the control group at 1 day after injury. Radial diffusivity analysis showed that this effect was mainly the result of acute edema. Pigs were euthanized at 5 days, and the brain tissues of xenon-treated animals showed reduction of perivascular inflammation and blood-brain barrier disruption. Endothelial cell culture experiments showed that glutamate reduces tight junction protein zona occludens-1 (ZO-1), but treatment with xenon microbubbles attenuates this effect. Xenon treatment protects cerebrovasculature and reduces astroglial reactivity after TBI. Further, these data support the future use of localized delivery of various therapeutic agents for brain injury using microbubbles in order to limit systemic side effects and reduce costs.