Search Results (1,284)

Precise control of dissolved oxygen is essential for reproducing physiologically relevant conditions in microfluidic cell culture systems. Here, we present a standalone, polydimethylsiloxane-free gas exchange unit (GEU) which enables controlled oxygenation and deoxygenation of perfused liquids and is suitable for integration with existing microfluidic platforms. The GEU employs a flat-sheet membrane contactor design to achieve efficient gas–liquid mass transfer while remaining independent of the downstream device. Oxygen transfer was experimentally characterised using optical oxygen sensors under different liquid and gas flow conditions. Reoxygenation efficiency decreased with increasing liquid flow rate because of reduced residence time, whereas active airflow through the gas compartment enhanced oxygen transfer. Controlled deoxygenation was achieved by flowing nitrogen through the gas compartment, with higher nitrogen pressures producing progressively lower oxygen levels. Numerical flow simulations demonstrated uniform flow distribution within the device, and a simplified analytical diffusion model accurately predicted the observed oxygen transfer trends. The proposed GEU provides a simple, robust and modular strategy for regulating dissolved oxygen upstream of microfluidic devices without requiring device redesign or specialised fabrication. This approach offers a practical solution for incorporating physiologically relevant oxygen control into a wide range of microfluidic and cell culture applications. Full article

Organoid and organ-on-chip technologies are rapidly evolving platforms for viral research that integrate stem cell biology, tissue engineering, and microfluidics to recapitulate key structural, mechanical, biochemical, and cellular features of human and animal physiology. By incorporating multicellular organoids into perfused microfluidic systems, these models can provide complex, dynamic, and physiologically relevant micro-environments for investigating virus–host interactions that are difficult to capture in conventional two-dimensional cultures and static organoids. Controlled flow, shear stress, extracellular matrix organization, tissue–tissue interfaces, and multicellular signaling enable mechanistic investigation of viral infectivity, dissemination, tissue injury and immune activation. Integration of real-time imaging and biosensors further permits longitudinal monitoring of viral replication, host responses, and tissue integrity, expanding the potential of these platforms for antiviral drug discovery. Recent organoid-on-chip studies using brain, skin, vaginal, respiratory, and intestinal models have demonstrated how tissue architecture, mechanical forces, glycocalyx dynamics, and immune–stromal interactions influence viral tropism and pathogenesis. In this review, we provide a mechanistic and translational overview of organoid and organ-on-chip technologies for studying viral infections, with particular emphasis on models of herpes simplex virus (HSV)-mediated disease. We further examine advances in immune integration, multi-organ systems, biosensing, and computational approaches that are expanding the complexity and predictive potential of these models. Importantly, patient-derived organoids and organ-on-chip platforms can capture interindividual differences in viral susceptibility, host responses, and therapeutic efficacy, providing pharmaceutical research with more precise, patient-relevant data to support drug prioritization and precision antiviral medicine. Finally, we discuss key barriers to broader adoption, including organoid maturation, biological and technical variability, reproducibility, scalability, biosafety, cost, standardization, and regulatory validation. Collectively, these advances position organoid and organ-on-chip technologies as powerful human-relevant models that bridge reductionist in vitro systems and human disease, while continued optimization, standardization, and validation will be essential to realize their full potential for mechanistically informed antiviral discovery, therapeutic development, and precision medicine. Full article

Prostate cancer remains a major clinical challenge, partly due to tumour–immune interactions that contribute to immune evasion and therapeutic resistance. The CD47–SIRPα axis is a key macrophage-associated immune recognition pathway; and materials-oriented platforms that allow preliminary investigation of tumour–macrophage interaction patterns remain valuable. Here, we report an optically traceable biohybrid nanoplatform based on CD47 aptamer-functionalized fluorescent carboxyl-functional MMES-POSS hybrid nanoparticles. The nanoparticles were synthesized within 5 min using UV-induced free-radical emulsion polymerization in an ethanol–water system and exhibited spherical morphology, SEM-derived dry-state mean diameters below 100 nm, negative surface charge, and retained fluorescence due to RITC encapsulation within the crosslinked hybrid matrix; however, DLS measurements revealed pronounced aggregation and polydispersity in aqueous dispersion. SEM and EDX provided complementary evidence of nanoparticle morphology and elemental composition, whereas zeta potential and Nanodrop measurements provided evidence consistent with surface functionalization, and FTIR confirmed retention of the core POSS, carbonyl, and RITC-associated chemistry. In a PC-3/THP-1 macrophage co-culture model, Annexin V–FITC/PI flow cytometry showed a concentration-dependent redistribution of cell populations, with the most pronounced early apoptotic enrichment observed at 2 µg/mL. However, pairwise exploratory comparisons among concentrations did not reach statistical significance, and this is discussed as a limitation of the present exploratory dose–response characterization. These findings indicate preliminary interaction-associated response patterns rather than definitive receptor-specific targeting or therapeutic CD47–SIRPα blockade. Overall, this modular POSS-based biohybrid interface provides a materials-oriented platform for probing prostate cancer–macrophage interaction profiles and guiding future mechanistic validation studies. Full article

Chitosan and calcium phosphate (CaP) are attractive bioactive coating materials for titanium (Ti) orthopedic implants, but coating approaches must provide uniform deposition, adequate adhesion, and cytocompatibility. This proof-of-concept study evaluated whether CaP microshells could be incorporated into electrosprayed chitosan coatings bonded to silanized Ti substrates without compromising coating properties. CaP microshells were synthesized using carbon microsphere templates and added to chitosan electrospray solutions at 0.25, 0.5, and 1.0 wt% relative to chitosan. Electrospray parameters were adjusted, and coatings were characterized by scanning electron microscopy, energy dispersive X-ray spectroscopy, Fourier transform infrared spectroscopy, tensile adhesion testing, water contact angle measurements, and W-20-17 bone marrow stromal cell culture. Increasing capillary diameter and reducing pressure enabled stable deposition of uniform composite coatings containing up to 1.0 wt% CaP. CaP microshells were distributed across the coating surfaces and throughout the coating thickness. Silanization significantly increased coating adhesion compared with non-silanized Ti, while CaP incorporation up to 1.0 wt% did not significantly reduce bond strength. All coatings were hydrophilic and supported viable cell attachment and growth over five days. These findings support the feasibility of electrosprayed chitosan–CaP microshell coatings as adhesive, cytocompatible bioactive coating platforms for Ti implant materials. Full article

Cardiac-related pain, including angina pectoris and pain accompanying myocardial infarction, remains clinically important, and existing treatments are inadequate for some patients. Animal and primary-tissue studies provide important mechanistic evidence, but species differences and limited access to patient-matched human neuro-cardiac tissue constrain translation. This review synthesizes the following separate advances relevant to future human iPSC-based cardiac-pain modeling: molecular mechanisms that may be reconstructed in vitro; platforms ranging from two-dimensional cultures to proposed sensory-innervated three-dimensional and microfluidic systems; and potential applications in target validation, patient-specific modeling, and analgesic screening. We distinguish direct human iPSC evidence from findings obtained in animal, primary-cell, cardiac-only, autonomic-neuron, or non-cardiac pain models. We also discuss maturation, standardization, and regulatory challenges. iPSC technologies may complement existing models and support mechanism-focused, patient-stratified research, but direct validation in human sensory–neuron–cardiac systems remains limited. Full article

Sugarcane (Saccharum spp. hybrid) is the most important sugar crop. However, its genetic improvement is severely constrained by genotype-dependent regeneration recalcitrance and lengthy tissue culture cycles. Here, we established a simple, efficient, and genotype-flexible in planta transformation system using two-leaf-stage plantlets. By systematic optimization of three key parameters: Agrobacterium cell density (OD₆₀₀ = 0.5), dark incubation duration (2 weeks), and infection frequency (two rounds), we achieved a maximum transformation efficiency of 51.2%. The protocol was successfully applied to eight diverse sugarcane varieties, with transformation efficiencies ranging from 33.3% to 51.2%, demonstrating broad genotype applicability. Using this optimized system, we introduced Sh4CL13 into sugarcane. Transgenic lines exhibited a 3.25-fold increase in Sh4CL13 transcript levels and a 13.7-fold elevation in chlorogenic acid (CGA) content compared to controls. Feeding bioassays with Mythimna separata larvae revealed that transgenic lines significantly prolonged larval developmental duration, reduced pupal weight, and decreased adult emergence rates. This in planta transformation system bypasses tissue culture, offers a practical platform for functional genomics and molecular breeding in sugarcane and potentially other monocot crops. Full article

The adult liver exhibits remarkable regenerative capacity in vivo; however, primary hepatocytes (PHs), the principal functional cells of the liver, swiftly forfeit their proliferative potential and specialized hepatic functions when isolated from their native microenvironment and cultured in vitro. While PHs are the benchmark for studying hepatic physiology, xenobiotic metabolism, and toxicological responses, their rapid dedifferentiation resulting in the loss of hepatic functions significantly limits their further application. Recent studies suggest three converging strategies to address the challenge of long-term maintenance and expansion of PHs. First, defined chemical and growth factor-based protocols can temporarily induce hepatocytes into a proliferative, progenitor-like state, followed by a maturation phase that restores differentiated hepatic functions. In addition, the inhibition of signaling pathways linked to cellular stress responses and identity loss can postpone dedifferentiation and preserve drug-metabolizing activity for prolonged durations, thereby enhancing disease modeling and toxicology studies. Finally, three-dimensional (3D) culture platforms generally improve hepatocyte maturation and functional stability, but these are often not scalable due to matrix dependence, technical complexity, handling requirements, and cost. This review thoroughly examines innovative methodological advancements designed to facilitate the proliferation and prolonged viability of healthy PHs, focusing on their translational relevance and clinical applicability. Full article

In drug-induced liver injury (DILI) prediction field, animal models and in vitro cell models are most commonly used. However, animal models require long experimental timelines and may exhibit species-specific differences compared with humans, whereas conventional two-dimensional (2D) cell culture models lack cell-to-cell and cell-to-extracellular matrix (ECM) interaction. Liver organoid models and liver organ-on-a-chip can better simulate the human liver microenvironment; however, the construction of liver organoids requires a long cycle and high costs, while liver organ-on-a-chip systems demand specialized equipment and professional technicians. Herein, we selected the human C3A cell line, characterized by its low cost and facile culture conditions to establish an in vitro three-dimensional (3D) liver model. Briefly, C3A cells were embedded in Matrigel and cultured for 7 days to allow model maturation. Compared with their 2D-cultured cell model, the established 3D model exhibited elevated mRNA expression levels of drug-metabolizing cytochrome P450 enzymes (CYPs). Moreover, the model displayed robust expression of key hepatic biomarkers, as well as bile duct biomarkers. To evaluate the model’s applicability for DILI prediction, we performed toxicity assessments using a panel of six well-characterized hepatotoxicants and three non-hepatotoxic compounds. Notably, the 3D C3A model achieved a sensitivity of 83.3%, specificity of 100%, and overall accuracy of 88.9%. Furthermore, treatment of this model with chlorpromazine, a well-characterized cholangiotoxic agent, resulted in suppressed expression of the bile duct biomarker cytokeratin 19 (CK19) and bile salt export pump (BSEP), accompanied by impaired bile acid transport capacity. Taken together, this study provided a simple, low-cost, easy to culture, and more readily scalable 3D hepatic model in comparison with conventional 2D primary human hepatocyte (PHHs) models and other advanced 3D liver models. Notably, the model displayed dual hepatic and biliary characteristics, supporting predictions of both DILI and drug-induced bile duct injury. It provided a promising in vitro platform for assessing drug-induced hepatobiliary toxicity, with potential to reduce reliance on animal experiments and accelerate early-stage screening of novel pharmaceutical candidates. Full article

Umbilical cord mesenchymal stem cell-derived extracellular vesicles (UCMSC-EVs) hold strong promise for regenerative medicine, yet their intrinsic size heterogeneity remains a critical barrier to clinical translation, as it obscures molecular and functional specialization within bulk EV preparations. Here, we pioneer the application of the automated EXODUS platform to directly fractionate EVs from cell culture supernatants, resolving bulk UCMSC-EVs into three size-defined subpopulations. By integrating this platform with high-resolution mass spectrometry, we systematically characterize the molecular and functional landscapes of these UCMSC-EV size subpopulations. We demonstrate that EV size is tightly linked to distinct biogenetic origins, biomolecular corona composition, and functional programs: smaller EVs are enriched in exosome-associated proteins, ECM–glycan interfaces, and corona-associated molecules, and preferentially engage endocytosis- and phagosome-related pathways, whereas larger EVs exhibit ectosomal signatures. These findings identify EV size as a critical determinant of molecular architecture and biological function, providing insight into size-dependent EV heterogeneity and informing the rational design and optimization of UCMSC-EV-based therapeutic strategies. Full article

Cardiovascular diseases (CVDs) remain a leading cause of mortality worldwide, and ageing is strongly associated with progressive arterial stiffening. Age-related alterations in extracellular matrix (ECM) mechanics influence vascular smooth muscle cell (VSMC) behaviour, while sympathetic innervation represents an additional regulator of vascular homeostasis. However, how neural signalling interacts with ageing-associated mechanical conditions to regulate VSMC behaviour remains unclear. In this study, an in vitro sympathetic neuron–VSMC co-culture model was established to investigate neuro-mechanical regulation. Primary rat sympathetic neurons and A7r5 VSMCs were cultured on glass or polydimethylsiloxane (PDMS) substrates with defined stiffness (20 and 130 kPa), representing healthy and ageing-associated stiffened arterial environments, respectively. VSMC behaviour was assessed through analysis of cell area, proliferation, migration, cellular Young’s modulus (YM), and DNA damage marker γH2AX. Sympathetic neuronal co-culture was associated with reduced VSMC spreading and decreased γH2AX levels. Under the conditions tested, neural signalling exerted limited effects on cell proliferation and migration. In contrast, increased substrate stiffness promoted cell proliferation and elevated YM. Both neuronal input and substrate stiffness were associated with increased cellular YM. Together, these findings indicate that neural and mechanical cues may jointly influence VSMC behaviour within ageing-associated mechanical environments. This co-culture system provides a controllable platform for studying neuro-mechanical interactions in vascular biology. Full article

Gliomas, and glioblastoma in particular, remain difficult to model because molecular heterogeneity, diffuse invasion, blood–brain and blood–tumour barrier effects, immune suppression and repeated therapeutic escape converge in the same disease. Two-dimensional cultures, glioma stem cell (GSC) systems, acute tumour slices and animal models remain indispensable for mechanistic research, pharmacology and in vivo validation. Glioma organoids are complementary research platforms, not components of routine diagnostic or treatment procedures. This review links model construction, microenvironmental validation, treatment perturbation and evidence-graded interpretation. We compare patient-derived glioma organoids, GSC-derived organoids, brain organoid–glioma co-cultures, genetically engineered brain tumour organoids, and vascular-associated, immune-cell-containing and chip-based platforms. We distinguish phenotypic resemblance from physiological fidelity, tumour-intrinsic drug sensitivity from delivery competence, and proof-of-concept activity from demonstrated clinical utility. We also examine temozolomide resistance, radiotherapy, targeted and combination therapy, antiangiogenic treatment, tumour-treating fields, immune-cell therapy, oncolytic viruses, multi-omic quality control and prospective validation. Organoids should not substitute for animal models or clinical trials. Their most defensible role is to provide a patient-derived functional layer between mechanism, regimen ranking and molecular tumour-board interpretation, with claims limited by assay reproducibility, clinically achievable exposure and outcome linkage. Full article

Oral cancer, predominantly represented by oral squamous cell carcinoma (OSCC), remains a major global health burden due to its aggressive clinical behavior, high recurrence rates, and limited improvement in survival over recent decades. Despite advances in treatment modalities, patient outcomes remain poor largely due to late diagnosis, therapeutic resistance, and profound tumor heterogeneity. In particular, metabolic reprogramming has emerged as a central hallmark of oral carcinogenesis, enabling tumor cells to adapt to hypoxic and nutrient-deprived microenvironments while promoting proliferation, invasion, and treatment resistance. Traditional experimental models, including two-dimensional cell cultures and in vivo animal models, have provided important mechanistic insights. However, they fail to fully recapitulate the metabolic, structural, and cellular complexity of human tumors. Consequently, there is growing interest in more physiologically relevant platforms, such as three-dimensional spheroids, organoids, and patient-derived models, which better preserve tumor architecture and microenvironmental interactions. In parallel, multi-omics approaches are increasingly being integrated to dissect the molecular and metabolic complexity of oral cancer at unprecedented resolutions, while nanotechnology-based systems are emerging as promising tools for targeted drug delivery and improved therapeutic precision. This review discusses experimental models in oral cancer research, focusing on their ability to capture metabolic alterations and tumor heterogeneity. We further highlight their translational potential together with multi-omics integration and nanotechnology-based strategies for improving biomarker discovery, therapeutic stratification, and the development of more effective treatment approaches. Full article

Visceral leishmaniasis, caused by Leishmania donovani, is characterized by profound alterations in hepatic immune responses that promote parasite persistence. However, the lack of physiologically relevant in vitro systems limits investigation of tissue-level host–pathogen interactions. An immunocompetent three-dimensional hepatic organoid model incorporating bone marrow-derived macrophages was established to investigate liver-stage immune responses to L. donovani infection. Integrated cytokine profiling and transcriptomic analyses were benchmarked against infected mouse liver tissue. Macrophages acquired functional characteristics consistent with a Kupffer cell-like phenotype and contributed to immune homeostasis within the co-culture. Infection induced a conserved immune response characterized by inflammatory and chemotactic activation. This response was accompanied by coordinated suppression of antimicrobial pathways, including the IL-12–IFN-γ axis and interferon-stimulated genes. These immune alterations were largely shared between the organoid model and infected liver tissue, although systemic metabolic reprogramming and multicellular complexity were not fully recapitulated. Our findings identify a conserved pattern of hepatic immune dysregulation in which inflammatory activation is uncoupled from effective antimicrobial signaling, potentially facilitating parasite persistence. This immunocompetent hepatic organoid platform provides a physiologically relevant system for investigating host–pathogen interactions and evaluating host-directed therapeutic strategies for visceral leishmaniasis. Full article

Yeast-assisted mixed-culture fermentations have gained attention for their ability to enhance fermentation efficiency and modulate metabolite production. There is little knowledge on the impact of magnetic nanoparticle-assisted immobilization on yeast–yeast interactions during alcoholic fermentation. The co-culture platforms of Saccharomyces cerevisiae and Kluyveromyces marxianus, immobilized on chitosan-coated manganese ferrite nanoparticles, were applied in the fermentation of sugarcane molasses and sugarcane juice in the present study. The chitosan-coated MnFe₂O₄ nanoparticles were prepared using a one-step coprecipitation reaction followed by hydrothermal treatment and were characterized by X-ray diffraction, Fourier transform infrared spectroscopy, vibrating sample magnetometry, and scanning electron microscopy. An immobilized co-culture system has been shown to provide faster sugar consumption and higher ethanol production than non-immobilized-cell fermentations. Operational stability of the immobilized biomass and higher ethanol production by reuse were confirmed by repeated fermentation cycles. Bioproduction of volatile metabolites varies among monoculture, co-culture, non-immobilized-cell, and immobilized systems, with yeast interactions and magnetic immobilization also affecting secondary metabolite production during fermentation. The findings confirm that magnetic nanoparticle-assisted co-culture fermentation would be an attractive nanobiotechnological tool for enhancing alcoholic fermentation and shed new light on yeast interactions with the nanostructured system when combined with the immobilized solution. Full article

Brain cancer is a highly aggressive disease with limited treatment options, highlighting the need for reliable preclinical models for drug discovery. This study aimed to isolate and characterize Saudi patient-derived primary brain cancer cells and assess the anticancer activity of novel compounds developed in-house. Sixteen tumor samples from Saudi patients were processed to establish primary brain cancer cultures and One Normal Tissue (Control). The cells were successfully isolated and maintained under optimized conditions, with their morphology and growth characteristics monitored. Molecular analysis confirmed the expression of key tumor and neural markers. The anticancer activity of selected compounds, KCO69, KCO70, and KCO129, was tested at various concentrations using the MTT and CellTiter-Glo Luminescent Cell Viability Assay. All compounds caused a concentration-dependent reduction in cell viability, with the strongest effects seen at 25 µM. Among them, compound 70 showed the most significant antiproliferative activity, while compounds KCO69 and KCO129 exhibited moderate effects. Variability in treatment response among cultures reflected the inherent heterogeneity of patient-derived tumors. Overall, establishing primary brain cancer cell models from Saudi patients offers a valuable platform for preclinical drug screening and supports further research on these compounds as potential therapies for brain cancer. Full article