Metal-organic frameworks (MOFs) are applicable in many fields; however, the high cost and toxicity of solvents make scale-up difficult. This study developed a novel, environmentally friendly, one-step synthesis of a Zeolitic Imidazolate Framework (ZIFL) and the in situ decoration of silver nanoparticles (AgNPs) using a Deep Eutectic Solvent (DES) at room temperature for pesticide residue detection. The DES acts as both a solvent and a stabilizing agent, facilitating the formation of the leaf-like ZIFL structure while ensuring the uniform dispersion of AgNPs on its surface. By exposing numerous coordination-unsaturated metal sites, the 2D ZIFL structure synergizes with AgNPs to achieve high catalytic efficiency, driven by the nanoparticles’ elevated surface charge density. Simultaneously, the strong affinity for thiol groups enables the ZIFL/Ag material to selectively bind Thiram, thereby increasing assay reliability in complex environments. Thiram absorbed at the surface of ZIFL/Ag blocks catalytic sites, causing the color of TMB to change from blue (oxidized state) to colorless. Using the proposed approach for Thiram detection, the LOD was achieved at 40 nM in DI water, with a linear range of 0–1 µM, and the corresponding correlation coefficient (R2) was 0.9773. Moreover, to demonstrate high on-site detection in low-resource settings, ZIFL/Ag-coated cotton swabs were introduced and used to detect Thiram in 5 min with the naked eye, combined with RGB analysis, thereby eliminating the need for spectroscopy. The study established results that demonstrate the potential to apply and integrate image processing to create simple, user-friendly, on-site devices with high accuracy for monitoring water quantity.
Microalgae are promising feedstocks for high-value compounds and biomass production, but harvesting remains a major challenge due to the high capital and energy costs, contamination risk, and low efficiency of conventional methods. In this study, we developed a novel spiral inertial microfluidic device with varying cross-sections to enable efficient concentration of microalgae across a broad size range. The device was designed to combine variable cross-sections with an expanded channel area to improve particle focusing and separation. Its performance was first evaluated using particles spanning a wide size range (3–20 μm) and then demonstrated using two representative microalgae species with distinct morphologies: Haematococcus pluvialis (spherical, 20 μm) and Phaeodactylum tricornutum (non-spherical, 3 μm in width). The device achieved excellent focusing performance for model particles, while recovery efficiencies for microalgae reached > 95
Extracellular vesicles (EVs), particularly exosomes, are gaining attention as next-generation therapeutic platforms. They serve as biologically derived nanocarriers capable of delivering functional molecules such as microRNA (miRNA). Exosomes possess high biocompatibility and low immunogenicity, protecting unstable nucleic acid therapeutics and enabling their efficient delivery to target cells. Consequently, technologies for manufacturing and engineering miRNA-loaded exosomes/EVs to deliver therapeutically relevant miRNAs are rapidly advancing. To ensure the therapeutic efficacy and reproducibility of miRNA-loaded exosomes, systematic analysis and characterization, including size, morphology, concentration, surface markers, miRNA loading efficiency, and stability, are essential. These analytical techniques are recognized as key elements enabling quality control and standardization of exosome-based therapeutics. Advancements in these manufacturing and analytical techniques have accelerated therapeutic applications for inflammatory diseases, with atopic dermatitis (AD) being a prime target. AD is a chronic inflammatory skin disorder characterized by skin barrier impairment and Th2-dominant immune dysregulation, where existing treatments face limitations in restoring long-term immune homeostasis. Recent studies indicate that miR-124, miR-146a, miR-143, miR-147a, miR-159a, and miR-223 contribute to improving AD pathophysiology by regulating inflammatory cytokine signaling and immune modulation pathways. Collectively, these findings highlight the therapeutic potential of miRNA-loaded EV platforms for the multi-target regulation of inflammatory skin diseases. This review focuses on the manufacturing and analytical techniques for miRNA-loaded exosomes/EVs, discussing the latest research trends in their potential therapeutic application for AD and challenges for future clinical translation.
(Background) Organ-on-a-Chip (OoC) technology has emerged as a microphysiological platform that addresses key limitations of conventional two-dimensional (2D) cell cultures and animal models in pharmacokinetics research. Reliable prediction of drug absorption, distribution, metabolism, and excretion (ADME) remains a major bottleneck in drug development, contributing to high clinical attrition rates driven by unanticipated toxicity and poor translational relevance. (Scope) OoC systems integrate human cells within dynamically perfused, three-dimensional microenvironments that recapitulate key structural, biochemical, and mechanical features of native organs. This review examines the mechanistic principles and pharmacokinetic applications of OoC platforms across major ADME-relevant organs—including the liver, kidney, gut, lung, and blood–brain barrier—and discusses their integration with physiologically based pharmacokinetic (PBPK) modeling. (Key Insights) Microfluidic flow, tissue–tissue interfaces, and mechanical cues sustain organ-specific functions and enable quantitative assessment of drug metabolism, transport, clearance, and tissue-specific toxicity. Representative studies demonstrate that OoC models capture human-specific drug responses overlooked by animal studies and generate pharmacokinetic parameters with close concordance to clinical observations. Multi-organ OoC configurations serve as physical analogues of systemic pharmacokinetic models, enabling interrogation of inter-organ crosstalk and whole-body drug disposition in vitro. (Concluding Perspective) Despite these advances, significant challenges related to biological fidelity, standardization, scalability, workflow integration, and regulatory qualification persist. Continued progress in automation, patient-derived models, artificial intelligence integration, and computational modeling is expected to further position OoC technologies as indispensable tools for predictive, human-relevant pharmacokinetics research and translational drug development.
Melanoma is a highly metastatic malignancy in which tumor-derived exosomes play critical roles in pre-metastatic niche formation and metastatic progression. However, most conventional therapeutic strategies primarily target primary tumor cells while overlooking metastasis-promoting exosomes. In this study, we developed a dual-targeting photothermal nanoplatform composed of Sambucus nigra agglutinin (SNA)-functionalized gold nanorods (SNA-AuNRs) capable of simultaneously recognizing melanoma cells and melanoma-derived exosomes through α-2,6 sialic acid targeting. SNA-AuNRs were synthesized by conjugating SNA lectin onto gold nanorods using PEGylation and EDC/NHS coupling chemistry, followed by physicochemical and biological characterization. The synthesized SNA-AuNRs retained strong near-infrared (NIR) absorbance and exhibited efficient photothermal conversion, reaching approximately 53 °C under 850 nm NIR irradiation for 10 min. In vitro studies demonstrated enhanced binding affinity of SNA-AuNRs toward melanoma cells and melanoma-derived exosomes compared to normal cellular controls. Notably, SNA-AuNRs achieved approximately 51.3
Muscle atrophy, a condition characterized by an imbalance between protein synthesis and degradation, leads to skeletal muscle wasting, exacerbating disease progression and increasing mortality rates. Exercise has been shown to counteract muscle wasting, with myokines—exercise-induced secretome proteins, and among them, irisin is known to mediate systemic benefits such as improved metabolism, reduction of oxidative stress, and promotion of tissue repair in skeletal muscle, thereby mediating beneficial effects of exercise against atrophy. Despite its promising therapeutic potential, the precise functions and signaling pathways of irisin in muscle and other organs remain insufficiently explored owing to the complexity and heterogeneity of in vivo environments. In this study, we present engineered skeletal muscle tissue models as biomimetic platforms to study irisin-mediated therapeutic interventions. Using this platform, we validated the ability of irisin to mitigate dexamethasone (DEX)-induced muscle atrophy by restoring myogenic markers, enhancing AKT-mediated protein synthesis, suppressing ubiquitin–proteasome–driven degradation, and preserving contractile function under catabolic stress. (e.g., irisin pretreatment increased tetanic stress in DEX-treated tissues by 43
Accurate and timely detection of Influenza A virus (H1N1) is essential for effective public health management. Traditional PCR-based methods, though effective, are limited by the need for thermal cycling and are impractical in point-of-care settings. To overcome this, we developed a novel isothermal detection strategy that utilizes the structure-specific activity of flap endonuclease 1 (FEN1) enzyme and the high signal-to-background contrast of a light-up RNA aptamer. In our system, the presence of H1N1 target RNA initiates the formation of a three-way junction (3WJ) structure, subsequently recognized and cleaved by FEN1. The resulting cleavage products then act as primers for a cascade of amplification reactions, leading to the transcription of light-up RNA aptamers. This label-free and amplification-driven method enables rapid and sensitive H1N1 detection with a low limit of detection (LOD) of 516.84 fM. Analysis based on three technical replicates (n = 3) demonstrated high reproducibility with coefficients of variation (CVs) consistently below 10
The global spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has highlighted the urgent need for rapid, sensitive, and field-deployable diagnostic platforms capable of detecting viral antigens prior to clinical intervention. Although localized surface plasmon resonance (LSPR) biosensors based on gold nanoparticles (AuNPs) offer label-free detection and simple optical readout, their sensitivity remains limited for detecting low-abundance viral proteins at early stages of infection. This study aimed to enhance the detection sensitivity of cuvette-based LSPR biosensors by introducing a dual-layer AuNP architecture that leverages the cumulative optical response of two plasmonically active layers for the ultrasensitive detection of SARS-CoV-2 nucleocapsid protein (NP). A cuvette-integrated dual-layer AuNP LSPR sensor chip was fabricated by vertically assembling two plasmonically active AuNP-coated substrates with a fixed separation distance. Antibody–antigen interactions were monitored through LSPR plasmonic peak shifts using absorbance-based spectrophotometric measurements. Sensor performance was evaluated in phosphate buffer and artificial human saliva, and analytical metrics including limit of detection (LOD), selectivity, coefficient of variation (CV), and recovery were systematically assessed. The dual-layer AuNP LSPR sensor exhibited a 2.5-fold improvement in detection sensitivity compared with a conventional single-layer configuration, achieving an LOD of 9.7 pM (0.78 ng·mL⁻¹) for SARS-CoV-2 NP with excellent linearity (R² > 0.99, n = 5). The sensor demonstrated high selectivity against non-target viral proteins and common interfering biomolecules. In artificial human saliva, the assay showed excellent analytical precision and accuracy, with CV values of 5.07–7.29
Metal–organic frameworks (MOFs) have been explored as drug delivery vehicles due to their structural tunability. Physical characteristics of MOFs including particle size and shape strongly influence cellular interactions and uptake. It is crucial for the rational design of drug carriers to optimize these parameters. Recent studies have reported that iron (Fe)-based MOFs can induce ferroptosis in cancer cells, that we hypothesized that their physical properties may directly affect ferroptotic efficiency. In this study, we synthesized four types of Fe-based MOFs (MIL-88(Fe)-NH2) with distinct shapes (octahedral and rod-shaped) and sizes (200 nm and 800 nm). Although all MIL-88(Fe)-NH2 exhibited the same crystalline structure and comparable specific surface areas, Rod-shaped MIL-88(Fe)-NH2 ( 200 nm in size) exhibited approximately 4-fold higher cellular uptake after 2 h compared to octahedral counterparts (n = 3, p < 0.0001), resulting in 2-fold intracellular Fe accumulation (n = 3, p < 0.01) and a 1.5-fold elevation in lipid peroxidation (n = 4, p < 0.0001, at 50 µg/mL). These results may be associated with their smaller size, higher aspect ratio, and smaller contact angle, which together promote efficient cellular internalization. In contrast, large-sized MIL-88(Fe)-NH2 exhibited comparable ferroptotic responses regardless of their shape. These findings provide new insights into the shape- and size-dependent behavior of Fe-based MOFs and offer a rational strategy for the design of MOF-based nanocarriers for cancer therapy. Shape and size-controlled iron (Fe)-based MOFs enable accelerated intracellular Fe delivery and potent ferroptotic cancer cell death. The 200 nm sized- and rod-shaped MIL-88(Fe)-NH2 exhibit faster cellular uptake and stronger ferroptosis induction than octahedron-shaped MIL-88(Fe)-NH2
Orally administered drugs undergo a complex process of absorption-distribution-metabolism-excretion (ADME), passing through the gut, liver, and kidney. Studying the ADME process of drugs can help predict the drug’s safety and pharmacokinetic properties. Orally administered drugs are absorbed in the gut and metabolized in the liver, and eliminated mostly through the kidney, where drugs are first filtered in the glomeruli and selectively reabsorbed in the tubules. Previously, we showed that a gut-liver chip can reproduce the dynamic and continuous process of gut absorption and liver metabolism. Here, a multi-organ system, Gut-Liver-Kidney (GLK) chip, integrating the main functions of the gut, liver, and size-selective renal filtration with physiologically realistic fluidic connection was developed to reproduce the ADME of orally administered drugs. We confirmed that the filtration and the reabsorption of model drug was reproduced in our GLK chip, mimicking the fate of drugs in the body. Our GLK chip can be a useful in vitro model system for studying the continuous process of ADME and consequent pharmacokinetics (PK) properties of oral drugs.
Lung disease is a major global public health challenge characterized by high mortality and significant diagnostic complexity due to overlapping clinical symptoms among diseases with distinct etiologies. Conventional diagnostic methods, such as chest imaging and sputum culture, often suffer from insufficient sensitivity for early detection or prolonged results that delay therapeutic intervention. As a promising alternative, aptasensors have emerged as powerful next-generation diagnostic tools due to their high target selectivity and structural stability. This review comprehensively summarizes recent advancements in aptasensor technologies for diagnosing major lung diseases, including pneumonia, tuberculosis, lung cancer, and asthma. It systematically analyzes research trends focusing on target biomarkers, signal transduction mechanisms, and functional nanomaterials while providing a comparative perspective on design strategies tailored to the distinct etiological mechanisms of each disease. Furthermore, this review addresses current technical hurdles such as matrix effects and long-term stability. Prospective directions for developing comprehensive multi-marker detection panels integrated with advanced nanomaterials and automated analytical systems are proposed to facilitate the molecular interpretation of complex pulmonary pathophysiology. This review provides a comprehensive overview to aid researchers in establishing next-generation diagnostic platforms that seamlessly integrate with clinical workflows.
Mechanobiology has evolved from macroscopic anatomical studies to a precise molecular understanding of how cells sense and respond to physical forces. While conventional tools like atomic force microscopy and traction force microscopy established the field, they often face trade-offs between force sensitivity and high-throughput spatial mapping in living cells. This review explores the transformative rise of DNA-based single-molecule force sensors, which leverage the programmability and defined physical attributes of nucleic acids to bridge this gap. We critically analyze the design principles and mechanical characteristics of four primary sensor classes: DNA duplexes, DNA hairpins, DNA origami, and DNA G-quadruplexes. Special attention is given to their fabrication and tuning for detecting piconewton-scale events. Furthermore, we highlight versatile applications ranging from dissecting cellular mechanotransduction to high-resolution bioimaging and biosensing. Finally, we discuss recent advances, projecting a future era of “mechano-medicine” where active DNA nanodevices profile mechanical phenotypes for diagnostic and therapeutic innovation.
Efficient intracellular delivery of therapeutic biomolecules, including messenger RNA, plasmid DNA, and CRISPR–Cas9 systems, is critical for advancing cell-based therapies and genome editing. However, conventional approaches, such as lipid-based nanocarriers and electroporation, often suffer from low cell viability, limited efficiency, and poor scalability. Microfluidic droplet-based cell squeezing offers a promising alternative by transiently permeabilizing membranes, enabling high delivery efficiency with reduced cytotoxicity and reagent consumption. Nevertheless, existing implementations are susceptible to clogging, which compromises operational reliability and reproducibility. Here, we present a z-axis droplet-squeezing platform that reconfigures the constriction geometry from a horizontal to a vertical orientation, effectively preventing clogging. This design maintains stable mechanoporation, facilitating efficient, carrier-free delivery of diverse biomolecules, with efficiencies reaching > 98
Hierarchical extracellular matrix (ECM) cues spanning mechanics, architecture, and matrix–matrix interfaces (MMIs) regulate the directionality and efficiency of tumor-cell migration and invasion. Despite their relevance, the contributions of interfacial structures within 3D ECMs remain under-resolved, particularly whether discrete boundaries serve as “attractor-and-track” drivers. Here, we engineered a polydimethylsiloxane (PDMS) microfluidic platform to create controlled MMIs that emulate the tumor microenvironment (TME)’s mechanical heterogeneity, achieved by sequential collagen gelation to create both planar and curved boundaries. With the system, we quantified how MDA-MB-231 (invasive) and MCF-7 (non-invasive) breast cancer spheroids migrate in uniform matrices of graded stiffness and when encountering soft–stiff boundaries. MDA-MB-231 spheroids demonstrated significantly greater migration in soft matrices and exhibited distinct invasive outgrowth at interfacial boundaries, with soft-top pairings (o–o, t–o) gating early detachment, followed by convergence of outgrowth across stiffness pairings at later times. Spheroids positioned above or below planar or curved MMIs showed directional approach toward the boundary and alignment of trajectories within the interfacial plane, consistent with interface-seeking and interface-parallel migration. In contrast, MCF-7 spheroids displayed minimal migration under all tested conditions, underscoring a phenotype-dependent responsiveness to ECM cues. Overall, our findings highlight the critical role of interfacial structures, in addition to bulk stiffness and architecture, in shaping cancer invasion, supporting a two-phase model in which local bulk mechanics license early outward dissemination, whereas interfacial stiffness increasingly sustains expansion. The proposed microfluidic platform offers a tunable and physiologically relevant model for dissecting 3D cell migration mechanisms within complex ECM environments, with optical access, curvature control, and validated passive gradients enabling future chemotaxis studies.
Bacterial surface display (Autodisplay) enables efficient selection of antibody fragments, but conventional phage display cannot express variable heavy (VH) and light (VL) chains separately, limiting combinatorial diversity and direct affinity-based screening. This study aimed to develop a combinatorial twin-chain Fv (tcFv) library enabling co-autodisplay of VH and VL chains on the surface of E. coli, and to enhance binding affinity through structural refolding. The Fv fragments derived from a human anti-thyroid peroxidase antibody were expressed independently using the autodisplay system. Randomized CDR3 regions of both chains were used to construct combinatorial libraries (genetic diversity: 1.52 × 10⁷ cfu). Screening was performed with Alexa 488-labeled erythropoietin (EPO) by flow cytometry and fluorescence-activated cell sorting. tcFv formation was confirmed by fluorescence resonance energy transfer (FRET) between Alexa 488–labeled VH and Alexa 546–labeled VL. Refolding was optimized using an oxido-shuffling reagent containing reduced and oxidized glutathione (GSH/GSSG, 10:1 ratio). A variant, tcFv.K18.23, exhibited high affinity toward EPO (KD = 49.2 nM, n = 3). FRET confirmed structural assembly of VH.K18 and VL.23 within < 10 nm proximity. Optimization of the VH:VL mixing ratio to 44
The abuse of psychoactive drugs such as ketamine and 3,4-methylenedioxymethamphetamine (MDMA) is increasing worldwide, and polysubstance abuse involving their concurrent use has become increasingly prevalent. Reliable on-site testing for both substances is therefore essential. However, conventional immunoassay platforms, including widely used commercial kits, remain limited in sensitivity and specificity for ketamine and MDMA compared with other drug metabolites, often leading to false negatives or cross-reactivity issues. To address these shortcomings, we introduce a microfluidic immunoassay-based on-site test device and validate its performance using certified reference materials and authentic urine samples. Authentic urine samples (N = 143), including drug-free controls (N = 45), ketamine-positive cases (N = 68), and MDMA-positive cases (N = 30), were evaluated. Verification results showed strong correlations (R2 = 0.87 and R2 = 0.90) between gas chromatography-mass spectrometry (GC-MS) data and the outputs of the developed device. Simultaneous detection revealed co-occurrence of ketamine and MDMA in 19.1
Extracellular vesicles (EVs), including exosomes, have emerged as powerful biomarkers and therapeutic vectors due to their molecular content and accessibility in biofluids. However, reliable EV isolation remains a significant challenge, particularly in clinical settings where high purity, low volume, and rapid processing are essential. This review systematically examines current EV isolation techniques, focusing on both conventional approaches such as ultracentrifugation, immunoaffinity capture, and size-exclusion chromatography and the rapidly evolving landscape of microfluidic technologies. Special attention is given to the two primary categories of microfluidic isolation: affinity-based and label-free systems. Affinity-based microfluidics utilize surface markers and ligand interactions to selectively isolate EV subpopulations with high specificity but face trade-offs in throughput and marker dependence. In contrast, label-free microfluidic platforms exploit physical properties like size, electrical charge, and acoustic responsiveness to enable gentle, high-throughput separation without biochemical labeling, preserving vesicle integrity and avoiding selection bias. Through a comparative evaluation of isolation performance, sample requirements, and scalability, this review highlights how microfluidic technologies address the core limitations of traditional methods. Furthermore, it discusses hybrid systems that integrate affinity and label-free mechanisms for enhanced efficiency and flexibility. We conclude by exploring future directions in automation, parallelization, and point-of-care integration, emphasizing the role of microfluidic EV isolation in advancing precision diagnostics and translational research. Together, these insights underscore the need for continued innovation toward standardized, clinically adaptable EV isolation platforms.
Hybrid nanoparticles (HNPs) have high potential in various biomedical fields, such as drug delivery, bio-signal detection, and delivery systems, as they can simultaneously realize biocompatibility and functionalization based on the complex structure of organic and inorganic materials. Microfluidic technology was introduced to synthesize HNP precisely to regulate size, shape, and functionality uniformity. Case studies of on-chip HNP platforms are presented, including microfluidic vascular models, tumor microenvironments, and blood-brain barrier models that enable real-time analysis of nanoparticle dynamics, drug release kinetics, and cellular uptake efficiency in physiologically relevant contexts. The review comprehensively examines the microfluidic-based synthesis of HNPs with the recent trends and possibilities of on-chip bio-application technology. In addition, diagnostic platforms and biosensor technologies using HNPs as detection mediators are discussed as biomarker detection, multiple signal response, and real-time monitoring technologies based on high sensitivity and selectivity. Integrated cell chip-based platforms enable simultaneous HNP analysis, streamlined sample preparation, and long-term biological interaction modeling, critical capabilities for advancing precision medicine. Therefore, it suggests that these technologies develop into an integrated platform that can contribute to precision medicine, self-diagnosis, and customized treatment in the near future.
Metal-organic framework (MOF)-based nanomaterials have attracted significant attention as active elements in electrochemical sensors. Although some progress has been made in the preparation of pure MOF electrodes, their low sensitivities render electrochemical glucose sensing difficult. Accordingly, in this study, a novel concept of incorporating a benzoic acid (BA)-based supporting linker into a Ni-based MOF (Ni-MOF) was proposed to achieve highly sensitive non-enzymatic glucose sensors. Addition of the BA-based supporting linker enhanced the ratio of the high oxidation state of Ni, thereby increasing the number of active sites for glucose detection. Introduction of 10 wt
Despite pharmacological-, technological- and medical- advances, cardiovascular diseases (CVDs) remain the main cause of death and disability in the world. This underscores the urgent need to better understand the early stages of these diseases for effective prevention, as well as to develop patient-specific pharmacological approaches (personalized medicine) and novel therapies. Traditional in vitro and in vivo models often fail to accurately mimic human physiology, limiting their translational potential. In this context, microphysiological systems (MPS) have emerged as advanced in vitro platforms that integrate key physiological features of cardiovascular tissues. This review summarizes the state-of-the-art advancements in in vitro models for studying CVDs, with a particular focus on emerging 3D cardiac and vascular models. These models serve as essential tools for disease modelling, drug development, and toxicity testing. Key parameters to consider when developing cardiovascular MPS are highlighted, along with a discussion of the advantages and challenges associated with each model system.
