Advanced Materials Technologies

Blood-based biomarkers are central to diagnostics, yet current approaches depend on invasive sampling and centralized laboratory infrastructure. At the same time, womens reproductive health remains severely under-monitored: most clinically relevant biomarkers are rarely measured outside fertility clinics, leaving millions without accessible, continuous insight into their reproductive lifespan. Anti-Mullerian hormone (AMH), a key indicator of ovarian reserve and overall reproductive function, still requires venous blood collection and specialized analysis, creating a major barrier to early detection, routine monitoring, and population-level screening. Here, we present a lateral flow assay (LFA) enabling direct AMH detection in unprocessed menstrual blood. The assay uses covalently conjugated 150 nm gold nanoshells to achieve sensitive colorimetric detection within the clinically relevant 0-10 ng/mL range. Results can be visually interpreted by naked-eye detection or quantified via a smartphone-based machine-learning algorithm for semi-quantitative assessment. The LFA performance correlates strongly with clinical chemistry lab-based analyses and can be seamlessly integrated into point-of-care formats, including wearable menstruation pads as well as simple dipstick tests. This technology provides a non-invasive, affordable, and robust solution for decentralized, regular monitoring of ovarian health.

The ability to rapidly fabricate custom polydimethylsiloxane (PDMS) devices is central to advancing organ-on-a-chip (OoC) technologies and other biological microplatforms. However, traditional photolithography and the surface roughness of directly 3D printed molds limit their accessibility and scalability of PDMS-based systems. Photolithographic workflows are limited by their dependence on specialized equipment, technical expertise, dedicated fabrication infrastructure, and are typically restricted to planar geometries and microscale features, limiting their use for millifluidic or complex 3D device features. To address these challenges, we present a modular workflow for the robust fabrication of PDMS-based devices using stereolithography (SLA) or fused deposition modeling (FDM) printing combined with optimized epoxy coatings. Acetone-thinned epoxy formulations dramatically improve SLA printed mold smoothness, eliminate tearing during demolding, and yield PDMS replicas with clean, well-defined structural features. For FDM printed molds, a two-step epoxy coating strategy restores mold quality sufficient for robust replica molding. The resulting PDMS devices support irreversible glass bonding, fluid containment, and cell culture applications, validated using normal mammary epithelial and cancer cell lines. We further demonstrate the formation of perfusable tissue aggregates within 3D matrices and introduce a low-cost 3D printed imaging platform for parallel live-cell imaging across four PDMS devices, showcasing its use for monitoring 20 OoC channels under gravity- or pump-driven flow. This versatile and reproducible method lowers the barrier to entry for soft lithography, allowing researchers without prior microfabrication expertise to rapidly prototype functional PDMS devices for diverse biological applications. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=102 SRC="FIGDIR/small/645830v1_ufig1.gif" ALT="Figure 1"> View larger version (44K): org.highwire.dtl.DTLVardef@bbf403org.highwire.dtl.DTLVardef@191249forg.highwire.dtl.DTLVardef@1fb7bf2org.highwire.dtl.DTLVardef@1865e4c_HPS_FORMAT_FIGEXP M_FIG C_FIG

Microphysiological systems (MPSs) have shown great promise for the advancement of drug discovery and toxicological tests, and as an alternative to animal models. However, although several chips and systems have been reported, some important issues are yet to be addressed, such as the use of polydimethylsiloxane (PDMS). Cyclo olefin polymers (COPs) have advantages over other thermoplastic materials, but most COP-based MPSs use solvent bonding during fabrication, which can affect any cells they are used to culture. This study uses a photobonding process with vacuum ultraviolet (UVU) to produce MPSs without the need for solvents such as cyclohexane, dichloromethane, and toluene. This is then used for comparison to investigate the effects of solvents on cell cultures. Quantitative immunofluorescent assays show that the coating efficiencies of extracellular matrix proteins, such as Matrigel and collagen I, are reduced on solvent-treated COP surfaces, compared with those prepared using VUV photobonding. Furthermore, SH-SY5Y neuroblastoma cells are used to evaluate cytotoxicity. This shows that solvent-MPSs induce apoptosis, but VUV-MPSs do not. These results provide insights into solvent bonding for MPS fabrication so that undesirable reactions can be avoided. Moreover, this work may be used to standardize MPS protocols and establish good manufacturing practices.

We report the fabrication of very thin microfluidic active and passive devices on rigid and flexible substrates for sample-space-restricted applications. Thin glass coverslips are commonly used substrates, but these being fragile often crack during experiments, leading to device failure. Here, we used PET as a flexible substrate to fabricate robust thin devices. We proposed a simpler process for PET-PDMS bonding without any silane, adhesive, and/or plasma treatment. We presented the compatibility of the thin devices with a digital in-line holographic microscope (DIHM) as a use case. The substitution of the conventional microscope with DIHM in microfluidic large-scale integrated systems renders simplicity, cost-effectiveness, portability, and miniaturization of the overall system. It also enables a customized and parallel multisite optical observation for a complex microfluidic circuit chip. These chips comprise various microfluidic components made of active microvalves, particularly Quake valves. We also successfully demonstrated the function of microvalves fabricated with our method to regulate the fluidic flow. Thus, are suited to making sophisticated microfluidic circuit chips to fit a variety of applications like organ-on-chip, cell culture, wearable biosensors, pressure sensors, etc.

Elastic porous membranes are essential components of mechanically active organ-on-a-chip and microphysiological system (MPS) platforms, where cyclic strain is required to recapitulate physiologically relevant tissue mechanics. However, existing fabrication methods are often difficult to reproduce, low throughput, or dependent on specialized infrastructure, limiting their adoption across laboratories. Many protocols also lack quality control steps for ensuring device assembling and reproducibility. In this paper, we present a robust and accessible fabrication and quality control workflow for the consistent production of elastic porous PDMS membranes. The method uses commercially available heat presses, release liners, and pre-patterned membrane wafers to enable rapid membrane molding. We describe a quality control framework, including visual verification of porous regions and wettability testing for surface activation, to ensure irreversible PDMS bonding and reliable device assembly. Together, this workflow improves fabrication yield, reduces device failure, and supports reproducible implementation of elastic porous membrane in organ-on-a-chip applications.

Light-based additive manufacturing methods have been widely used to print high-resolution 3D structures for applications in tissue engineering, soft robotics, photonics, and microfluidics, among others. Despite this progress, multi-material printing with these methods remains challenging due to constraints associated with hardware modifications, control systems, cross-contaminations, waste, and resin properties. Here, we report a new printing platform coined Meniscus-enabled Projection Stereolithography (MAPS), a vat-free method that relies on generating and maintaining a resin meniscus between a crosslinked structure and bottom window and to print lateral, vertical, discrete, or gradient multi-material 3D structures with little-to-no cross-contamination or waste. We also show that MAPS is compatible with a wide range of resins and can print complex multi-material 3D structures without requiring specialized hardware, software, or complex washing protocols. MAPSs ability to print structures with microscale variations in mechanical stiffness, opacity, surface energy, cell densities, and magnetic properties provides a generic method to make advanced materials for a broad range of applications.

Despite technological advances, the fabrication of multiscale, multi-material, and topologically complex 3D structures using soft hydrogel bioinks remains a challenge due to the inherent trade-offs between print size/resolution, bioink properties, and design complexity. In this work, we combine additive (macroscale) digital light projection (DLP) mode with subtractive (microscale) two-photon ablation (TPA) mode with multi-material exchange capability. We identify ideal hydrogel bioink formulations that are compatible with both DLP and TPA modes of processing. Technical challenges related to multimodal fabrication such as alignment of multiscale topologies to facilitate seamless media perfusion, soft-hard multi-material printing to facilitate handling of mechanically weak hydrogel constructs, and hydrogel swelling during printing, were resolved. To highlight the novelty of this hybrid platform, we fabricated centimeter-scale bioink constructs with embedded microscale perfusable topologies that cannot be achieved by isolated use of either DLP or TPA modes. This includes simpler microfluidic chips with independently perfusable microchannels to more complex 3D constructs with embedded, multiscale, perfusable dual-fluidic circuits that mimic the alveoli-capillary interface, or microfluidic chips with endothelialized microchannels. The unique ability of this multimodal platform to mimic in vivo-like multiscale complexities can be potentially used to develop next-generation organ-on-chips.

Active implantable microscale reservoir-based drug delivery systems enabled novel and effective drug delivery concepts for both systemic and localized drug delivery applications. These systems typically consist of a drug reservoir and an active pumping mechanism for precise delivery of drugs. Here we present a stand-alone, refillable, scalable, and fully implantable microreservoir platform to be integrated with micropumps as a storing component of active implantable drug delivery microsystems. The microreservoir was fabricated with 3D-printing technology, enabling miniature, scalable, and planar structure, optimized for subcutaneous implantation especially in small animals (e.g., mouse), while being readily scalable for larger animals and human translation. Three different capacities of the microreservoir (1 L, 10 L, and 100 L) were fabricated and characterized all with 3 mm thickness. The microreservoir consists of two main parts: a cavity for long-term drug storage with an outlet microtubing (250 m OD, 125 m ID), and a refill port for transcutaneous refills through a septum. The cavity membrane is fabricated with thin Parylene-C layers using a polyethylene glycol sacrificial layer, minimizing restoring force and hence backflow, as fluid is discharged. This feature enables integration to normally-open mechanisms and improves pumping efficiency when integrated to normally-closed pumps. The results of in vitro optimization and characterization of the cavity membrane show 95% extraction percentage of the cavity with insignificant (2%) backflow due to restoring force of the membrane. The refill port septum thickness is minimized down to 1 mm by a novel pre-compression concept, while capable of ~65000 injections with 30 Ga non-coring needles without leakage under 100 kPa (4x greater than physiological backpressure). To demonstrate integrability of the microreservoir to an active micropump, the 10 L microreservoir was integrated to a micropump recently developed in our laboratory, making an implantable drug delivery microsystem. Two different microsystems were subcutaneously implanted in two mice, and the outlet microtubing was implanted into the round window membrane niche for infusion of a known ototoxic compound (sodium salicylate) at 50 nL/min for 20 min. Real-time shifts in distortion product otoacoustic emission thresholds and amplitudes were measured during the infusion. The in vivo results show a mean shift of 22.1 dB after 20 min for the most basal region, matching with syringe pump results. A biocompatibility experiment was performed on the microsystem for six months to assess design and fabrication suitability for chronic subcutaneous implantation and clinical translational development. The results demonstrate very favorable signs of biocompatibility for long-term implantation. Although tested here on mice for a specific inner ear application, this low-cost design and fabrication methodology is scalable for use in larger animals and human for different applications/delivery sites.

1This work describes a versatile and cost-effective cell culture method for growing adherent cells on a porous membrane using pressure-sensitive double-sided adhesives. This technique allows cell culture using conventional methods and easy transfer to microfluidic chip devices. To support the viability of our system, we evaluate the toxicity effect of four different adhesives on two distinct airway epithelial cell lines and show functional applications for microfluidic cell culture chip fabrication. We showed that cells could be grown and expanded on a "floating" membrane, which can be transferred upon cell confluency to a microfluidic chip for further analysis. The viability of cells and their inflammatory responses to IL-1{beta} stimulation was investigated. Such a technique would be useful to culture cells in a conventional fashion, which is more convenient and faster, and stimulate cells in an advanced model with perfusion when needed.

Cardiovascular diseases cause an estimated 17.9 million deaths globally each year (World Health Organization). Endothelial cells that line the vasculature and the endocardium are subjected to cyclic mechanical stretch. Deviation from physiological stretch can alter the endothelial function, having the risk of atherosclerosis and myocardial infarction. To understand the mechanical stretch effects, cell culture platforms that provide mechanical stretch have been developed. However, most of them have fixed strain and frequency, sometime not in the pathophysiological range. We thus developed a novel, electromagnetically driven, uniaxial stretching device, where cells were grown on a flexible polydimethylsiloxane (PDMS) membrane mounted onto a 3-D printed track. The strain of the membrane was readily controlled by tailoring the track design and the frequency was determined by electromagnetic actuation. Furthermore, the mechanical strain gradient was generated on a PDMS membrane with a tapered thickness. This strain gradient, ranging from 1.5% to 40%, covered both physiological and pathological vascular stretch ranges. When human vascular endothelial cells were subjected to the cyclic stretch, the cells exhibited strain-dependent cell and nuclear orientation and elongation perpendicular to the stretching direction, compared to the random cell and nuclear orientation under the static condition. However, the overstretching led to deviation from the aforementioned orientation and elongation, and impaired the tight junctions, leading to a leaky endothelium. This novel, versatile, cost-effective, pathophysiologically relevant stretching device provides a useful platform for advancement of vascular disease research and treatment.

High-throughput microfluidics has transformed biomedical research by enabling precise and parallel sample handling, but most devices are single-use due to channel occlusion and contamination from experiments. Alongside low fabrication yield and reduced experimental success associated with dense microfeatures, this creates a major bottleneck for scalable high-throughput applications. We present a rapid, reusable, and modular high-throughput microfluidic platform with integrated microvalves for automation. The platform employs a multilayer architecture consisting of a custom casing, PDMS layers with dense microfeatures for fluid handling and culture, and a glass substrate. Permanent bonding is applied only between control and fluid layers, while reversible bonding is used at all other interfaces, including the substrate. Because substrate is the primary cell-contact surface and can be readily detached, the remaining layers can be disassembled, thoroughly cleaned, and reused with minimal processing on a new substrate. This approach improves repeatability and experimental success while reducing preparation time from days to [~]2 hours. The disassemblable design also supports incorporation of application-specific layers between fluid layer and substrate, enhancing platform versatility for 3D culture. We validated performance through pressure/flow characterization and on-chip cell/organoid culture. Overall, our platform accelerates rapid high-throughput data generation across diverse biological applications.

Although recent wearable chronic wound (CWO) bandage technologies have opened up exciting opportunities for personalized CWO management, they still face significant obstacles due to the reliance on the wound bed exudate for sensing and delivering therapeutics. Flat, shallow, and desiccated wounds are difficult to collect wound exudate for sensing, and some wounds continuously exude, potentially washing delivered therapeutics out of the wound bed. Herein, we developed a hydrogel-forming microneedles (HFMNs) array-based multimodal transdermal dressing system that continuously monitors the on-site physiological conditions of CWOs in interstitial fluid (ISF) and offers healing capabilities. The unique polar array design enables the integration of six replaceable HFMNs sensing electrodes to target the desired wound-specific analytes in transdermal interstitial fluid (glucose, uric acid, pH, Na+, Cl-, K+, and temperature) based on their significance in reflecting the status of the CWOs. The hydrogel is composed of a biocompatible and swellable polymer - polyvinyl alcohol, and chitosan as a crosslinking agent, while the incorporation of MXene (Ti3C2Tx) nanosheets as conductive nanofillers facilitates the formation of 3D polymer hydrogel networks via hydrogen bonding. Further coating and functionalization of poly(3,4-ethylenedioxythiophene): polystyrene sulfate (PEDOT: PSS) and graphene oxide through a laser-scribed phase separation (LSPS) process improves the electrical conductivity and in-vivo water stability of the HFMNs as a result of the larger and interconnected PEDOT-rich domains. Importantly, anti-inflammatory and antibacterial properties of the hydrogel prevent wound infection and promote skin wound healing. Through the potential correlation between wound-affected ISF and wound bed exudate, this method bridges conventional and implantable dressing systems for commercialization.

Vat Photopolymerization (VPP) holds much promise for producing biomaterial constructs such as porous scaffolds. However, achieving micron-scale pore dimensions with precision presents a challenge. This study offers an innovative approach to stabilize the silicone elastomer vat surface permitting micron-scale layer thickness accuracy to be maintained. Internal and surface contamination on the poly(dimethyl siloxane) (PDMS) vat surface were observed and effectively controlled with a pre-saturation methodology, and porous structures with cubical pores were then printed with varying layer thicknesses. These structures demonstrate the ability to achieve micrometric resolution and layer thicknesses as fine as 32 {micro}m. A scaffold suitable for in vivo implantation with 40 {micro}m cubical pores was successfully printed within 5 hours using a stabilized PDMS vat surface. Additionally, the methodologys adaptability to intricate non-linear edge porous structures underscores its versatility across X, Y, and Z-axis.

The fabrication of microfluidic devices has progressed from cleanroom manufacturing to replica molding in polymers, and more recently to direct manufacturing by subtractive (e.g., laser machining) and additive (e.g., 3D printing) techniques, notably digital light processing (DLP) photopolymerization. However, many methods require technical expertise and while DLP 3D printers remain expensive at a cost [~]15-30K USD with [~]8M pixels that are 25-40 {micro}m in size. Here, we introduce (i) the use of low-cost ([~]150-600 USD) liquid crystal display (LCD) photopolymerization 3D printing with [~]8M-58M pixels that are 18-35 {micro}m in size for direct microfluidic device fabrication and (ii) a poly(ethylene glycol) diacrylate-based ink developed for LCD 3D printing (PLInk). We optimized PLInk for high resolution, fast 3D printing and biocompatibility while considering the illumination inhomogeneity and low power density of LCD 3D printers. We made lateral features as small as 75 {micro}m, 22-{micro}m-thick embedded membranes, and circular channels with a 110 {micro}m radius. We 3D printed microfluidic devices previously manufactured by other methods, including an embedded 3D micromixer, a membrane microvalve, and an autonomous capillaric circuit (CC) deployed for interferon-{gamma} detection with excellent performance (limit of detection: 12 pg mL-1, CV: 6.8%), and we demonstrated compatibility with cell culture. Finally, large area manufacturing was illustrated by printing 42 CCs with embedded microchannels in <45 min. LCD 3D printing together with tailored inks pave the way for democratizing access to high-resolution manufacturing of ready-to-use microfluidic devices by anyone, anywhere.

Organ-on-Chips (OoC) have the potential to revolutionize drug testing. However, the fragmented ecosystem of available OoC systems leads to wasted resources and collaboration barriers, slowing uptake. To address this, there is a need for OoC platforms based on interoperability standards, modularity, and reconfigurability. Technology platforms based on open designs would enable seamless integration of diverse OoC models and components, facilitating translation. Our study introduces a modular microfluidic platform that integrates swappable modules for pumping, sensing, and OoCs, all within the ANSI/SLAS microplate footprint. Sub-components operate as microfluidic building blocks (MFBBs) and can interface with the demonstrated Fluidic Circuit Board (FCB) universally as long as the designs adhere to ISO standards. The platform architecture allows tube-less inter-module interactions via arbitrary and reconfigurable fluidic circuits. We demonstrate two possible fluidic configurations which include in-line sensors and furthermore demonstrate biological functionality by running both in-vitro and ex-vivo OoC models for multiple days. This platform is designed to support automated multi-organ experiments, independent of OoC type or material. All designs shown are made open source to encourage broader compatibility and collaboration.

Bioxolography enables high-resolution fabrication of geometrically complex, cell-laden constructs for tissue engineering. However, tissue-relevant cell densities conflict with the optical transparency required for efficient dual-color volumetric printing. In this work, we extend the Bioxolography toolbox to include refractive index (RI) matching for cell-laden bioresins using iodixanol (IDX). Remarkably, IDX enhances optical transparency and boosts reactivity -- a phenomenon unique to Xolography. Yet, excessive IDX compromises dual-color efficiency through increased absorption and undesired UV-only curing, underscoring a central trade-off between optical clarity and photochemical performance. Systematic tuning of resin compositions along an iso-refractive index line demonstrated the versatility of Bioxolography, with IDX enhancing polymerization and 4-Hydroxy-TEMPO providing biocompatible inhibition. Optimizing composition and printing parameters yielded GelMA hydrogels with cell densities up to 5{middle dot}106 cells{middle dot}mL-1. Cell-laden prints achieved sub-100 {micro}m resolution and complex geometries such as channels and gyroids. Using skeletal muscle tissue as a model, we validated RI matched Bioxolography as a promising strategy for tissue engineering by demonstrating cell alignment along printed grooves and formation of mature muscle fibers characterized by MyoHC+ staining and fusion index. By integrating physical, chemical, and biological perspectives, this work advances Xolography toward biomaterials development and reinforces its position as an emerging volumetric (bio)printing technology. Table of Contents O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=175 SRC="FIGDIR/small/729865v1_ufig1.gif" ALT="Figure 1"> View larger version (86K): org.highwire.dtl.DTLVardef@407d86org.highwire.dtl.DTLVardef@1f116b4org.highwire.dtl.DTLVardef@109a437org.highwire.dtl.DTLVardef@1fc4ece_HPS_FORMAT_FIGEXP M_FIG C_FIG For printing higher cell density bioresins with Xolography, iodixanol (IDX) is added for refractive index-matching. The addition leads to an unexpected additional effect with increased reactivity in the dual-color photopolymerization. With careful adjustment of the resin composition and the printing parameters, Bioxolography is proven as a viable tool for tissue engineering.

Paper-based diagnostics such as lateral flow assays (LFAs) and microfluidic paper-based analytical devices ({micro}PADs) have attracted considerable attention because of their low cost, portability, and ease of use. Currently, to enable fabrication of {micro}PADs and improve LFA performance, hydrophobic blocks are patterned on paper substrates. However, fabrication of high-resolution hydrophobic barriers remains a major challenge. In this work, we developed a novel silicone extrudable ink for the fabrication of hydrophobic features on paper substrates. The ink was formulated using a vinyl-terminated polydimethylsiloxane (vPDMS) and polymethylhydrosiloxane (PMHS) system crosslinked through platinum-catalyzed hydrosilylation, and its rheological properties were tailored by incorporating silica fillers, obtaining a shear-thinning gel suitable for extrusion. The resulting formulation provided tunable properties, controlled deposition, and stable feature formation, enabling simple, low-cost, rapid, and robust fabrication of high-resolution hydrophobic barriers. Using this approach, we demonstrated improved fluid confinement and pattern fidelity on paper substrates, fabricated high-resolution paper microfluidic devices down to 150 {micro}m channel width, and enhanced the sensitivity of an LFA for a malaria diagnostic test. These results highlight the potential of this silicone ink platform as a practical and scalable strategy for advancing high-performance paper-based diagnostic technologies.

Currently in vitro models of microvascular biology rely on self-assembly of vascular cells in compatible gels. However, the stochastic nature of this process results in large variations in lumen sizes, perfusion continuity, and shear stresses making systematic and reproducible analysis challenging. Here, we report a new technology to generate artificial capillaries on a chip with custom control over lumen sizes and architectures using a combination of femtosecond laser cavitation and collagen casting within multi-chambered microfluidic chips. The design allows seeding of endothelial cells within capillary-sized microchannels and seeding of stromal cells within top-open silos, with independent control over seeding sequence and media compositions. Results show that endothelialized microchannels, coined as artificial capillaries, exhibit excellent barrier function with reproducible control over lumen sizes ({phi}=8-35{micro}m) and their architectures (straight, curvatures, tapered, branched). The physical flow parameters measured across the lumen (namely, flow shear) and at the channel outlets (flow velocities) have been validated against high-fidelity numerical assessments from the Large Eddy Simulation scheme within the digitized versions of the microchannels. The experiment-computation compatibility enabled us to predict changes in regional velocity and wall shear stresses within artificial capillaries, for various capillary architectures. We also show that in situ editing of artificial capillaries in the form of adding new branches or adding occlusions is possible. Lastly, we developed a co-culture model which enables the study of stromal cells with artificial capillaries using conventional imaging methods. We envision that acellular chips with two seeding ports can be readily shipped worldwide and could potentially be adopted as a new technology to study microvascular biology in a reproducible manner.

Oxygen availability is a critical yet all-too-often overlooked variable in organ-on-a-chip (OoC) systems. PDMS-based microfluidics remain the most common approach to facilitating oxygen equilibration with the incubator environment, but the materials tendency to ad- and absorb small hydrophobic molecules can pose significant concerns for pharmacological and toxicological studies. Yet there remains a lack of alternative gas-exchange materials feasible for OoC integration, even as the use of thermoplastic microfluidics in particular has otherwise proliferated. Here, we present commercially available track-etched nanoporous polycarbonate (50 nm pores, 1.18% porosity, [~]0.1 {euro}/cm2) as a practical alternative to polydimethylsiloxane (PDMS) for gas exchange in OoC. We show that nanoporous polycarbonate provides a thermoplastic material with an oxygen permeability of 3290 {+/-} 240 fs mol / kg, over an order of magnitude higher than PDMS. We demonstrate integration into existing lamination-based thermoplastic microfluidic fabrication workflows with sustained leak-free operation well above physiologically relevant pressures. We find that nanoporous polycarbonate does not compromise cell viability, but that high water vapor permeance necessitates a high-humidity environment around the device - though thickness-normalized water vapor permeability is notably similar to PDMS. We validate the OoC application with Caco-2 intestinal epithelial cells by monitoring oxygen levels during the critical cell attachment phase, with nanoporous polycarbonate allowing for maintenance of stable oxygen tension, in stark contrast to severe hypoxia in nonporous controls within 30 minutes. We further show that this uncontrolled hypoxia correlates with a time-delayed increase in cellular hypoxia inducible factor-1 reporter expression. Overall, our findings position nanoporous polycarbonate as a low-cost, mechanically robust, and fabrication-friendly alternative that can bring controlled oxygen availability to PDMS-free microfluidics and OoC.

We developed and characterized a three-layer microfluidic Exposure-on-a-Chip (EOC) model to replicate the alveolar microenvironment and simulate breathing motions, offering a physiologically relevant platform for studying inhaled nanomedicines. The EOC chip, fabricated from biocompatible polydimethylsiloxane (PDMS), features a fluidic chamber for cell culture, a pneumatic chamber for pressure application, and a thin PDMS membrane (150 {micro}m) separating the chambers to support cell growth and enable mechanical stretching. Xurography and 3D printing were validated as efficient and reproducible fabrication methods. Mechanical characterization, using fluorescent bead tracing, confirmed that the PDMS membrane accurately mimics alveolar breathing motions under physiological conditions (1-8% strain). Biological validation showed that alveolar epithelial and endothelial cells cultured on the EOC formed functional monolayers, maintaining barrier integrity under cyclic and static stretching, separately. To study nanoparticle behavior, we examined the deposition of nanoparticles under dynamic stretching versus static conditions. Significantly fewer nanoparticles accumulated in cells under continuous dynamic exposure with stretching compared to static culture, highlighting the critical role of mechanical forces in nanoparticle-cell interactions. The EOC platform provides a robust and scalable tool for evaluating nanomedicine efficacy in dynamic environments, representing a significant advancement in alveolus-on-chip technology.