Search Results (500)

Pulmonary diseases such as Chronic obstructive pulmonary disease (COPD), asthma, pulmonary fibrosis, and acute respiratory infections remain a major global health challenge due to their complex pathophysiology and limited therapeutic options. Conventional 2D cultures and animal models have provided foundational insights; however, they often fail to accurately replicate the human lung’s intricate architecture, immune interactions, and patient-specific variability. Recent advances in vitro technologies have transformed pulmonary research, enabling the generation of physiologically relevant and translational disease models. The review highlights the progression of lung research platforms from traditional monolayer cultures to advanced systems such as air–liquid interface models and 3D lung organoids. These cutting-edge models more effectively mimic the biochemical, mechanical, and spatial microenvironment of the respiratory system, enhancing the fidelity of disease modelling and drug screening. In parallel, the integration of computational modelling and artificial intelligence (AI) has emerged as a powerful synergistic approach. AI-driven analytics facilitate high-throughput imaging, biomarker discovery, and patient-stratified therapeutic prediction, while computational tools simulate disease networks, mechanobiological interactions, and pharmacological responses. The convergence of these technologies supports a deeper understanding of pulmonary disease progression and accelerates the development of precision therapeutics. Collectively, this review underscores the transformative potential of combining in vitro lung models with advanced computational and AI methodologies. This synergy not only improves translational relevance and reduces reliance on animal testing but also paves the way for personalised interventions that better address the complexity of human pulmonary disease. Full article

Cancer immunotherapy has transformed oncology, including the management of urothelial carcinoma, yet response rates remain limited and mechanisms of resistance are incompletely understood. At the same time, recent initiatives from the National Institutes of Health and the United States Food and Drug Administration have emphasized the development of standardized human organoid platforms to improve preclinical modeling and reduce reliance on traditional animal systems. Urothelial cancer provides a compelling context in which to advance these efforts, given its established responsiveness to Bacillus Calmette–Guérin and immune checkpoint blockade and its marked heterogeneity in clinical outcomes. Here, we review current organoid-based platforms for modeling bladder cancer immunotherapy, including patient biopsy-derived epithelial tumor organoids, air–liquid interface cultures that retain endogenous stromal and immune components, engineered bladder cancer assembloids that reconstruct defined multicellular circuits, and induced pluripotent stem cell-derived urothelial organoids that offer renewable multilineage systems. For each approach, we outline strengths, technical constraints, and limitations in scalability, immune fidelity, and genetic matching. We conclude by discussing the critical challenges that must be addressed—including benchmarking to patient tumors, reproducibility across laboratories, and standardized validation metrics—to enable regulatory acceptance and clinical translation of organoid-based immunotherapy testing. Full article

This review summarizes innovative co-formulation strategies for non-marketed dry powder inhalers (DPIs), enabling the simultaneous pulmonary delivery of multiple active pharmaceutical ingredients (APIs). Key approaches include co-amorphous systems (COAMS) and co-crystals, which combine two APIs into a single particle, improving aerodynamic properties, solubility, dissolution, and patient compliance while reducing manufacturing complexity. Core–shell microparticles, produced via spray drying, allow spatial separation and controlled release of APIs, minimizing drug–drug interactions and enabling tailored pharmacokinetics. Co-spray drying of dual APIs can yield particles with superior aerosolization and stability, though examples remain limited. Nanoparticle-based systems offer enhanced lung deposition and cellular uptake but face challenges in device compatibility, scalability, and regulatory approval. Each technology presents unique advantages and limitations regarding manufacturability, dose flexibility, and clinical translation. This review also highlights advances in in vitro toxicity testing, including air–liquid interface cultures, organoids, lung-on-chip models, and precision-cut lung slices, which are increasingly important as alternatives to animal studies. The importance of using an aerosol exposure system for the testing is highlighted. Ultimately, the choice of co-formulation platform should balance scientific innovation with practical considerations of manufacturing and regulatory requirements to maximize therapeutic benefit and commercial viability for future DPI combination products. Full article

The rational design and regulation of interfacial microenvironments represents an effective strategy for enhancing reaction performance. Previous studies have demonstrated that constructing air–liquid–solid triphase interfaces can substantially enhance catalytic reactions involving gaseous reactants. However, research on regulating the triphasic interfacial microenvironment remains limited and challenging. Herein, we fabricated a triphase photocatalytic system by depositing hydrophobic materials onto ordered TiO₂ porous (OTP), achieving significantly enhanced performance in visible-light-driven dye-sensitized photooxidation. Further, we regulated the triphasic microenvironment by systematically adjusting the chain length of hydrophobic molecules. It was found that the chain length greatly affects the interfacial properties, including O₂ concentration, the organic molecule adsorption and the interfacial electron transfer efficiency, thereby influencing photocatalytic reaction kinetics and pathways. We demonstrated a high-performance triphase photocatalytic system using 1H,1H,2H,2H-perfluorooctyl triethoxysilane as the hydrophobic material, which optimized multiple interfacial properties through synergistic effects, leading to optimal photocatalytic performance. Full article

Mitochondrial dysfunction drives persistent inflammation in severe asthma, yet its upstream metabolic regulation remains unclear. Induced sputum from patients with severe asthma was analyzed and integrated with transcriptomic datasets from independent cohorts. Two mouse models (C57BL/6J) were used for in vivo validation with multi-omics profiling, and mechanistic studies were performed in air–liquid interface-cultured primary human airway epithelial cells. Glutathione reduced form (GSHr) was markedly depleted in sputum and associated with poor disease control and mixed granulocytic inflammation in patients with severe asthma. Multi-omics analyses revealed coordinated disruption of glutathione (GSH) metabolism, including oxidized GSH accumulation, reduced synthesis and glutathione-S-transferase activity, and impaired mitochondrial GSH transport. GSH supplementation alleviated airway inflammation, oxidative stress, and mitochondrial dysfunction, whereas pharmacological inhibition of GST exacerbated these effects. Mitochondrial analyses identified suppressed SLC25A39 expression as a key mediator of defective GSH transport and redox imbalance. Transcriptomic profiling of airway biopsies showed upregulation of Neuropilin-1 (Nrp1), closely associated with altered glutathione pathways. Targeting the Nrp1 b1 domain restored mitochondrial GSH metabolism and attenuated airway inflammation. These findings identify an Nrp-centered metabolic pathway that disrupts mitochondrial homeostasis and drives inflammatory amplification, highlighting mitochondria-targeted therapeutic strategies for severe asthma. Full article

Chronic exposure to benzo[a]pyrene (BaP), a Group 1 IARC carcinogen, is a major driver of lung carcinogenesis; however, how sustained subcytotoxic exposure reprograms bronchial epithelium toward preneoplastic states remains poorly defined. Here, we subjected BEAS-2B human bronchial epithelial cells to 12 weeks of continuous BaP at environmentally relevant concentrations (0.1 and 1.0 µM) and interrogated the resulting phenotypes using an integrated multi-scale framework encompassing functional toxicology, RT-qPCR, RNA-seq, phospho-kinase/NF-κB arrays, and organotypic air–liquid interface (ALI) cultures. Cells maintained metabolic competence throughout, evidenced by sustained CYP1A1 and CYP1B1 induction at both acute (4 h) and chronic (12-week) timepoints, while accumulating genotoxic stress as demonstrated by dose-dependent nuclear γ-H2AX foci formation and ATM phosphorylation (Ser1981). RNA-seq revealed a dose-dependent transcriptional shift: 0.1 µM BaP yielded 119 differentially expressed genes (DEGs; |log2FC| ≥ 1, FDR < 0.05), whereas 1.0 µM generated 255 DEGs. Downregulated transcripts were enriched for extracellular matrix and cell-adhesion programs (COL14A1, ADAMTS2, CSMD3, CADM3), while upregulated genes encompassed inflammatory, calcium-signaling, and vesicle-trafficking modules (NFATC4, CSF2RA, SYT1, PCLO). Phospho-kinase/NF-κB arrays confirmed a p53/NF-κB signaling nexus, with concurrent activation of MAPK/ERK (Thr202/Tyr204) and PI3K/Akt (Ser473) pathways. Despite persistent genotoxic stress, cells did not acquire anchorage-independent growth and remained non-tumorigenic in vivo. Critically, ALI organotypic cultures derived from BaP-exposed cells exhibited histological dysplasia, nuclear pleomorphism, and disrupted apical-basal polarity. These findings mechanistically link chronic BaP exposure to an initiation-like preneoplastic state and establish a validated 2D/3D multi-omics platform for PAH-driven lung carcinogenesis research. Full article

Structural optimization of the cathode flow field is a viable approach to homogenize multi-physical field distributions and boost the output of air-cooled proton exchange membrane fuel cells (PEMFCs). This work develops a three-dimensional non-isothermal model to systematically evaluate the performance of graded flow channel designs. The results indicate that the graded structure promotes fluid transport in the central zone, thereby improving oxygen distribution uniformity at the gas diffusion layer/catalyst layer (GDL/CL) interface. Compared to the traditional parallel flow channel (with an average oxygen mass fraction of 0.051% and a uniformity index of 0.779), this configuration yields a 6.4% increase in the average oxygen mass fraction and a 0.96% enhancement in distribution uniformity. However, increased gradient flow reduces the flow velocity within the channels and raises the operating temperature, posing challenges for water and thermal management. The curved channel design, featuring longer channels at the ends and shorter channels in the center, compensates for the uneven air supply caused by the fan, thus balancing the flow distribution. Among the tested configurations, the 10° curved structure exhibits optimal performance, achieving the best compromise between gas distribution and liquid water removal. It effectively promotes oxygen diffusion and uniform water distribution, significantly alleviating mass transfer polarization and yielding a more uniform interface temperature distribution due to evaporative cooling. Both excessively small and large curvature angles lead to performance degradation, primarily due to inadequate water removal and flow separation, accompanied by excessive pressure drop, respectively. In contrast, the 10° curved channel strikes an optimal balance, offering significant advantages in overall cell performance and water–thermal management, which provides critical guidance for optimizing PEMFC flow field designs. Full article

The wake characteristics behind a transom stern vessel play a crucial role in determining its hydrodynamic performance, resistance, and environmental impact. This hydrodynamic phenomenon involves violent wave breaking, posing significant challenges for experimental analysis. In this study, we explore the complex wake dynamics behind a transom stern vessel using high-fidelity three-dimensional numerical simulations. A sharp volume of fluid method is employed to capture the gas–liquid interface, while the immersed boundary method is applied to simulate the ship hull boundaries. A distinct advantage of the present simulation is the capability to conduct quantitative analysis within the turbulent two-phase mixing region characterized by significant air entrainment, which is difficult for traditional experimental and theoretical approaches. The research focuses on the interaction between free surface dynamics, air entrainment and turbulent vortex structures, which collectively shape the wake region. The main flow features of wakes, including wave patterns across various Froude numbers, air entrainment and the evolution of bubbly wakes, are investigated. Furthermore, the correlation between turbulent vortex structures and violent interface breaking is examined. Full article

This pioneering study investigates bacteria isolated from marine pelagic fish, fishing vessels, and gear surfaces, focusing on the variability in biofilm formation across different substrates, media, and cultivation conditions. Bacteria from fish intestines, skin, and gills, including spoilage organisms and potential fish and human pathogens, can contaminate vessel surfaces, gear, and containers and may act as microbial reservoirs and transmission vectors. In this study, biofilm formation was evaluated at air–liquid interfaces and on submerged plastic, metal, and glass surfaces under various incubation temperatures and media. Vibrio spp. were isolated both from fishing nets and fish gills, particularly Vibrio alginolyticus, V. gigantis, and V. pelagius. Although V. harveyi was examined as a representative vibrio, it did not form a biofilm on polypropylene. Photobacterium damselae subsp. damselae, Pseudomonas fragi, P. gessardii, Psychrobacter spp., and Rothia endophytica showed a strong affinity for stainless steel. Overall adhesion regardless of media type was highest for P. gessardii, followed by P. damselae and Aeromonas veronii, which adhered strongly to steel, glass, and polypropylene; however, only P. gessardii also adhered well to polystyrene, an important finding because these are known fish and human pathogens. These results highlight species-dependent biofilm triggers and their substantial variability and provide guidance for standardized marine biofilm protocols. Full article

Gasoline engine exhaust (GEE) has been reported to contribute to the pathogenesis of pulmonary diseases. Autophagy, proinflammatory cytokines, and the NF-κB pathway core protein may play roles in the development of lung diseases caused by GEE. However, little is known about the possible toxic effects. Herein, we aimed to examine the crosstalk between GEE and the expression levels of autophagy-associated proteins (microtubule-associated proteins 1A/1B light chain 3A (LC3I/II)), proinflammatory cytokine genes (including interleukin-1β (IL-1β), IL-6 and IL-8), and the NF-kB pathway core protein p65 by conducting an air–liquid interface exposure study in BEAS-2B cells. A CCK-8 assay was conducted to explore the viability of BEAS-2B cells exposed to GEE and 3-methyladenine (3-MA). The protein expression levels of LC3I/II and p65 were detected using Western blotting. The gene expression levels of LC3B, IL-1β, IL-6, and IL-8 were measured using real-time PCR. We found that GEE decreased the viability of BEAS-2B cells in a dose-dependent manner, whereas 10%GEE exposure and 2.5 mM 3-MA had no significant effect. As the dose of GEE increased, LC3I/II protein and gene expression levels, proinflammatory cytokine gene expression levels, and p65 protein expression levels showed varying degrees of changes. Additionally, after treatment with 3-MA, these indicators tended to decrease, but only the gene expression levels of proinflammatory cytokines were statistically significant. These results suggest that GEE could interfere with autophagy and induce an inflammatory response in human bronchial epithelial cells, and that modest changes in autophagy could significantly alleviate this response, thereby providing new insights for the understanding of lung injury caused by GEE. Full article

The cystic fibrosis transmembrane conductance regulator (CFTR) is a member of the atypical ATP-binding cassette (ABC) family that functions as a phosphorylation-regulated epithelial anion channel. Cystic fibrosis (CF) is characterised by variants in the CFTR gene that lead to impaired epithelial chloride–ion transport and increased mucus viscosity. Although CFTR modulators such as Trikafta^® have transformed the care of many CF patients, individuals harbouring rare CFTR variants still have no effective treatment options. In this study, we used primary air–liquid interface (ALI) airway cultures obtained from 21 CF patients (pwCF) and 21 healthy controls (HC) to evaluate the therapeutic efficacy of CFTR restoration based on chitosan-mediated CFTR mRNA and modulators. While modulators restored CFTR channel function in most cultures derived from CF patients, those with class I or other rare variants showed no improvement. Chitosan-mediated CFTR mRNA delivery successfully restored CFTR function in ALI cultures of patients carrying rare CFTR variants with limited or no observed clinical response to modulator therapy, assessed by electrophysiology using our newly developed Multi Transepithelial Current Clamp (MTECC) Ussing chamber. This was then confirmed by morphological visualisation of CFTR protein expression in modulator-responsive patient samples using immunofluorescence (IF) staining. IF revealed an increase in CFTR signal and the restoration of epithelial barrier integrity following chitosan-mRNA and modulator treatment as a secondary outcome alongside CFTR functional measurements. Notably, MUC5AC expression, a major gel-forming mucin expressed by airway goblet cells and mucus viscosity were elevated in CF cultures, but were markedly reduced following successful intervention, approaching the levels seen in HCs. These findings establish the potential of chitosan-mRNA delivery as a therapeutic approach for CF patients, particularly those who do not respond to modulators. They also provide a practical, comparative evaluation of advanced mRNA-based treatments in patient-derived airway models. Full article

Safe drinking water and microbial inactivation from surfaces and devices are among the World Health Organization’s priorities. Plasma-activated water (PAW) inactivates microorganisms mainly by producing radicals (hydroxyl radicals, superoxide, nitrogen oxide, etc.), which form secondary reactive species like nitrates, nitrites, hydrogen peroxide, etc., from the air–liquid interface, where the plasma interacts with the water. A plasma arc device for water treatment with enhanced arc length was constructed at the Andronikashvili Institute of Physics (TSU) and used in the study. PAW’s antibacterial efficacy has been evaluated against Gram-negative E. coli and remarkably stress-resistant Gram-positive B. pumilus. This study identifies reactive oxygen (hydrogen peroxide and superoxide anions) and nitrogen species (total nitrate and nitrite ions) in plasma-activated water, analyzing their potential impact on antioxidant enzyme activity and their relationships with bacterial cell viability. B. pumilus exhibits greater resistance to plasma-activated water as a disinfectant compared to E. coli. Catalase is more effective than superoxide dismutase in protecting cells from external oxidative stress, based on the two antioxidant enzymes studied. Full article

Background: Pulmonary neuroendocrine cells (PNECs) are rare airway sensory cells implicated in amplifying allergic inflammation, yet due to their scarcity, the contribution of PNECs to the metabolic programs and responses of the airway epithelium remains poorly defined. Using a newly developed PNEC-enriched human airway epithelial model (ePNEC), we investigated the influence of PNECs on neuroendocrine and immune-modulatory metabolite production in response to the common aeroallergen of the house dust mite (HDM). Methods: Human bronchial epithelial cells (HBECs) and ePNEC cultures were differentiated at the air–liquid interface. Global untargeted metabolomics was performed to quantify metabolite abundance at baseline and following stimulation with HDMs. Differential expression, overlap significance, metabolite class enrichment, and pathway analyses were used to define PNEC-specific metabolic programs. Results: Principal component analysis (PCA) demonstrated strong baseline separation between ePNECs and HBECs, with HDMs inducing additional within-cell-type shifts. ePNECs displayed broader and more pronounced metabolite changes than HBECs. Baseline differences were largely preserved following allergen exposure, with significant overlap in both up- and down-regulated metabolites. ePNECs exhibited enriched neurotransmitter-linked metabolites—including serotonin, L-noradrenaline, dopamine, and histamine—at baseline and after HDM exposure. Amino acid–centered metabolism dominated the dataset, with enhanced histidine and tryptophan pathway activity in ePNECs. Pathway analysis revealed significant enrichment of phenylalanine, tyrosine, tryptophan, glutathione, and arginine–proline metabolism in ePNECs, whereas HBECs showed no significant pathway-level enrichment after HDM exposure. Conclusions: Human ePNECs engage a distinct, neuroactive metabolic program that is amplified upon HDM exposure. These findings provide a metabolic framework for how PNECs shape epithelial and neuroimmune responses to inhaled allergens. Full article

In two-phase flow, the interaction between multi-scale vortex structures and interfaces (bubbles or free surfaces) triggers a range of complex physical phenomena. This study employs numerical simulations to investigate the interaction between a horizontal vortex and the interface separating two layers of immiscible fluids with different densities (e.g., water and air). The vortex is initialized as an internal motion within the heavier phase. We focus specifically on the impact of the phase density ratio and surface tension. Numerical simulations reveal that when the density ratio is near unity, interface rupture occurs only at high Weber numbers ($\mathit{We}$), where low surface tension enables the rupture of sharp interface points. Conversely, at high surface tension (low $\mathit{We}$), these sharp points stretch into thin liquid films, significantly increasing the surface area without causing breakage. As the density ratio increases, interface rupture at sharp points accelerates, even under high surface tension, leading to faster dissipation of the initial vortex. In high-$\mathit{We}$ scenarios, an increased density ratio promotes the faster formation and greater intensity of new vortex layers at the interface. However, increasing surface tension enhances the vorticity of these layers but simultaneously slows their generation rate. The findings highlight the critical interplay between surface tension and density differences in vortex–interface interactions, with surface tension stabilizing the interface and density differences driving more intense vortex shedding and deformation. These insights offer valuable guidance for understanding two-phase flow behavior and improving the design of systems involving multiphase fluids. Full article

The development of effective inhalable drugs remains a key challenge in the treatment of pulmonary diseases, due to the physiological barriers of the respiratory tract and the lack of predictive models that accurately reproduce the human lung environment. In this context, liposomes (LP) have emerged as promising nanocarriers for pulmonary drug delivery due to their high biocompatibility, surfactant-like composition, capacity to encapsulate both hydrophilic and lipophilic drugs, and potential to provide sustained drug release while reducing systemic toxicity. This study evaluates the influence of size and PEGylation on their physicochemical properties, cytotoxicity, interaction with the pulmonary mucus, and cellular internalisation. LP of 100 nm (LP 100), 200 nm (LP 200), and 600 nm (LP 600) were characterised physiochemically and evaluated in pulmonary cell lines (A549 and Calu-3) exposed in liquid–liquid interface (LLI) and air–liquid interface (ALI) by nebulisation. In addition, artificial pulmonary mucus (APM) was employed to analyse LP penetration through the pulmonary mucus barrier. Results indicate that LP 100 exhibits greater colloidal stability, lower cytotoxicity, and sustained migration through the APM over time with respect to larger particles. PEGylation of LP 100 (LP-PEG) further increases their stability and ability to penetrate the APM, although cellular internalisation is reduced due to the steric effect of the PEG coating. These findings highlight the importance of adjusting the size and surface modifications of LPs according to the therapeutic target of the drug, optimising their persistence on the epithelial surface or their cellular uptake. Full article