Search Results (466)

Cancer metabolism is a dynamic and context-dependent process shaped by both tumor-intrinsic programs and microenvironmental cues. Capturing this complexity remains a major challenge, which limits the translation of metabolic insights into clinically actionable strategies. Patient-derived tumor organoids, together with emerging engineered platforms such as organ-on-chip systems, vascularized assembloids, and bioprinted tumor models, have opened new avenues for investigating tumor metabolism in physiologically relevant settings. These models enable the study of metabolic heterogeneity across tumor types, disease stages, and treatment conditions while preserving clinically relevant tumor features. Importantly, they provide functional platforms for ex vivo metabolic profiling, identification of metabolic vulnerabilities, and prediction of therapeutic responses. In this review, we discuss recent advances in the use of patient-derived and engineered 3D tumor models to characterize context-dependent metabolic states and treatment-induced metabolic rewiring. We first review technologies currently available to interrogate metabolism in these systems, including optical metabolic imaging, spatial metabolomics, isotope tracing, and bioenergetic profiling. We then discuss how 3D tumor models are used to investigate metabolic interactions within the tumor microenvironment (TME), including stromal and immune crosstalk, acidosis, nutrient availability, and circadian regulation. Finally, we critically examine current limitations, particularly the insufficient physiological relevance of standard organoid culture conditions for metabolic studies, and discuss how advanced engineering approaches and computational modeling may contribute to metabolism-driven functional precision oncology. Full article

Drug-induced liver injury (DILI) remains a major cause of acute liver failure and drug withdrawal from the market. Recently developed three-dimensional (3D) hepatic in vitro systems exhibit improved functionality and drug sensitivity compared with conventional two-dimensional cultures. These 3D models range from simple physiologic-like culture systems to advanced bioreactors with dynamic flow to provide sufficient nutrients and consistent drug exposure. However, whether dynamic perfusion improves sensitivity and reproducibility of hepatotoxicity testing remains unclear. Here, we developed a tailor-made perfusion platform to support volumetric bioprinted hepatic constructs for hepatotoxicity testing. The constructs consist of intrahepatic cholangiocyte organoids (ICOs) differentiated towards hepatocyte lineage and embedded in a gelatin methacryloyl bioresin. For toxicity evaluation, the hepatocyte-like ICO constructs were exposed to prolonged subtoxic acetaminophen treatment (10 mM, 7 days). The perfusion system effectively maintained and enhanced hepatocyte differentiation, evidenced by upregulated hepatic markers under perfused conditions compared to static controls. Testing of acetaminophen hepatotoxicity revealed that the perfused constructs displayed elevated cellular injury, with markedly higher liver injury markers relative to controls. Collectively, this study demonstrates the successful application of perfusion-based 3D model culture and highlights its potential as a more physiological platform for hepatotoxicity risk assessment in drug discovery and regenerative medicine. Full article

To preliminarily elucidate the bidirectional metabolic effects caused by changes in PFKM expression, provide research clues for further investigation of the molecular mechanisms through which PFKM regulates porcine skeletal muscle metabolism and myogenic differentiation, and offer a reference for identifying candidate genes associated with meat production traits and studying gene regulation in pig breeding, this study compared skeletal muscle protein expression profiles between Large White and Bama pigs. Candidate regulatory factors related to muscle growth and energy metabolism were screened, and the effects of altered muscle-type phosphofructokinase (PFKM) expression on metabolic homeostasis and myogenic differentiation in porcine skeletal muscle satellite cells (SMSCs) were preliminarily evaluated. Longissimus dorsi muscle tissues from Large White and Bama pigs were analyzed using iTRAQ-based proteomics. A total of 2040 reliably quantified proteins were identified, of which 51 were relatively upregulated in Large White pigs and 73 were relatively upregulated in Bama pigs. Functional enrichment analysis showed that the differentially expressed proteins were mainly involved in glycolysis, mitochondrial energy metabolism, protein synthesis, and the regulation of muscle fiber structure and function. PFKM was therefore selected as a key candidate differentially expressed protein. Porcine SMSC models comprising a PFKM knockdown group (PFKM-KD), a PFKM overexpression group (PFKM-OE), and a normal control group (PFKM-CON) were subsequently established. Glucose consumption and lactate accumulation in the culture medium, ATP levels, reactive oxygen species (ROS), mitochondrial membrane potential, apoptosis, mitochondrial dynamics-related proteins, and myogenic differentiation markers were then examined. Compared with the PFKM-CON group, the PFKM-OE group showed significantly increased glucose consumption and lactate accumulation, together with significant increases in ROS levels, mitochondrial membrane potential, and apoptosis, whereas ATP levels were significantly reduced. In the PFKM-KD group, glucose consumption and lactate accumulation were significantly decreased, accompanied by reductions in mitochondrial membrane potential, ROS, ATP levels, and apoptosis. PFKM overexpression mainly induced oxidative stress, ATP depletion, and increased apoptosis, whereas PFKM knockdown primarily reduced mitochondrial membrane potential, ROS, and ATP levels, indicating a relatively low-metabolic state. Both treatments were accompanied by dysregulated expression of the mitochondrial dynamics-related proteins DRP1, MFN2, and OPA1, although their patterns of change were not identical. Western blotting and immunofluorescence consistently showed that the expression levels of the myogenic differentiation markers MyoD and MYH were significantly lower in both the PFKM-KD and PFKM-OE groups than in the control group, suggesting that either excessive or insufficient PFKM expression may impair the myogenic differentiation potential of SMSCs. In conclusion, changes in PFKM expression are closely associated with glycolysis-related metabolism, energy and redox homeostasis, mitochondrial function-related indicators, and myogenic differentiation capacity in porcine SMSCs. The normal biological function of PFKM may therefore depend on its expression being maintained within an appropriate range. Full article

Given the urgent need to regulate oxidative stress microenvironments in chronic wound healing, hydrogel dressings that simultaneously integrate antioxidant, antibacterial, mechanically adaptive, and biocompatible properties are highly desirable. In this study, a natural polymer-based composite hydrogel dressing loaded with zinc-quercetin nanozyme (Zn-Q) was designed. The gel skeleton was constructed via a dual network of photocrosslinked methacrylated silk fibroin (SilMA) and mercaptolated hyaluronic acid (HA-SH) via thiol-ene click chemistry, with the catalase (CAT)-like Zn-Q nanozyme encapsulated in situ within the network, thereby achieving synergy between chemical crosslinking and dynamic metal-polyphenol coordination. Systematic characterization revealed that Zn-Q nanozyme adopted a stable octahedral coordination configuration, and its continuous porous structure exposed abundant catalytically active sites. The composite hydrogels exhibited a highly interconnected, three-dimensional (3D) porous morphology, with swelling ratios that increased significantly with Zn-Q nanozyme content (up to around 1082%). Rheological and mechanical tests demonstrated that although incorporating the nanozyme reduced the storage modulus, the reversible physical crosslinks formed via hydrogen bonding and coordination interactions endowed the material with excellent tensile toughness and energy-dissipation capacity, exhibiting typical Mullins softening behavior. Functional evaluation showed that Zn-Q nanozyme conferred superior free radical scavenging capability to the hydrogels and exerted dose-dependent inhibition against both Staphylococcus aureus and Escherichia coli. Furthermore, the hydrogels exhibited favorable adhesion to various wet organs and heterogeneous material surfaces, with hemolysis rates below 5% and cell viability exceeding 100% after 3 days of culturing with fibroblasts, confirming their excellent hemocompatibility and cytocompatibility. This study provides an experimental basis for developing a new type of wound repair materials that integrate antioxidant, anti-infective, and mechanically adaptive properties, holding significant application potential in oxidative stress-related tissue repair fields. Full article

Plant secondary metabolites remain indispensable for pharmaceuticals, nutraceuticals, and cosmeceuticals, yet conventional plant culture systems are increasingly limited by inconsistent yields, poor scalability, and inadequate capacity for real-time process monitoring. Microfluidic technologies and organ-on-a-chip (OoC) platforms, originally developed for mammalian biology, are now emerging as powerful tools to overcome these constraints. These systems enable laminar flow, precise gradient generation, single-cell resolution, and biosensor integration, providing unprecedented control over the cellular microenvironment and supporting non-destructive, real-time metabolomic monitoring. While recent reviews have surveyed plant microfluidics broadly covering developmental biology, single-cell phenotyping, and root–microbe interactions, this review provides, to our knowledge, the first synthesis focused specifically on organ-on-a-chip approaches for plant secondary metabolite biosynthesis and real-time metabolomic monitoring. Advances in device fabrication, including PDMS, paper-based, hydrogel, and thermoplastic materials, surface engineering, gradient-based elicitation strategies, and integration of optical, electrochemical, and mass spectrometric detection systems have also been critically examined. Special emphasis is placed on root-on-a-chip, shoot meristem, protoplast, callus, and 3D organoid platforms for studying cell wall mechanics, vacuolar dynamics, cytoskeletal responses, and signalling cascades. However, challenges remain in long-term culture stability and scalability; nonetheless, these technologies offer a roadmap toward programmable ‘plant biosynthetic factories’ to produce high-value natural products. Full article

Background: Subclinical mastitis is a major challenge in dairy production, contributing to antimicrobial use, production losses, and persistent infection pressure while often remaining undetected due to the absence of clinical signs. Using somatic cell count (SCC) data to distinguish newly infected from chronically infected cows is epidemiologically important, yet little is known about how infection dynamics relate to bacterial diversity in these groups. The present study aimed (i) to quantify the herd-level association between newly and chronically subclinically infected cows based on SCC, and (ii) to compare the abundance distribution of culturable bacteria in these groups using ecological diversity indices. Methods: We combined longitudinal Dairy Herd Improvement (DHI) data from 88 Danish dairy herds with quarter-level microbiological culture and MALDI-TOF MS identification from 1738 cows. Results: A strong positive herd-level correlation was observed between newly and chronically infected cows (Spearman’s r = 0.76, p < 0.001), indicating substantial shared variation without implying causality. Both Simpson (D) and Shannon–Wiener (H′) indices differed significantly between newly and chronically infected cows, demonstrating distinct bacterial abundance distributions. Non-aureus staphylococci dominated both groups, whereas Staphylococcus aureus and Streptococcus uberis were markedly more prevalent in chronically infected cows. Conclusion: the findings show that newly and chronically subclinical mastitis represent distinct epidemiological and ecological states, underscoring the need to combine SCC data with species-specific diagnostics to support effective herd-level mastitis control. Full article

Colorectal cancer (CRC) exhibits a highly heterogeneous tumor immune microenvironment (TME), ranging from “immune-inflamed” to “immune-desert” or “immune-excluded” phenotypes. Understanding how immune cell composition, cytolytic activity (CYT) and genomic alternations shape tumor-immune interactions is critical for improving immunotherapy outcomes. We analyzed TCGA-COAD and TCGA-READ datasets to evaluate immune competency, CYT, immune subtypes, microsatellite instability (MSI), and genomic instability, including somatic mutations, copy number aberrations (CNAs), and chromothriptic events. Immune cell infiltration was correlated with CYT levels, immune checkpoint expression, and immune-related gene signatures. Immune-competent (IC) tumors were predominantly CYT-high, enriched in stromal and immune scores, and exhibited distinct TME characteristics compared with immune-deficient (ID) tumors. IC/CYT-high tumors expressed higher levels of immune checkpoints (PD-1, PD-L1, CTLA-4, IDO1/2, LAG-3) and cytokines/chemokines (C1QA/B/C, CXCL9/10/11, CXCL13). Differences in immune infiltration were observed across tumors with significant mutations and copy number alterations. No prognostic difference was observed between CYT-high and CYT-low patients, indicating that CYT reflects immune activation rather than clinical outcome. Functionally, stimulated CD8⁺ T cells exhibited cytotoxicity activity against MSI-high (HCT-116) and microsatellite-stable (HT-29) CRC cells, with MSI-H cells showing higher sensitivity. Dynamic 3D co-culture demonstrated tumor-guided T cell infiltration and retention of CD8 expression, and co-culture was associated with moderate upregulation of cytotoxicity-related genes GZMA and PRF1 within the system. Cytotoxic activity decreased at lower effector-to-target ratios, highlighting the importance of effector dose. Overall, these findings link CYT, immune competency, MSI status, and genomic instability to T cell cytotoxic responses, providing insights into tumor-immune interactions, and suggest potential associations relevant for immunotherapy research in CRC. Full article

Bladder cancer is the fifth most common cancer in the United States, causing approximately 17,000 deaths annually. Due to its vast genetic and molecular heterogeneity, presentation, prognosis, and therapeutic response vary greatly between individuals. To improve patient outcomes, there is a need for better drug-screening platforms. The genetic heterogeneity of bladder cancer often leads to chemotherapy resistance or low response rates. Moreover, chemotherapies are often contraindicated in patients with select comorbidities. Organoids offer a better option to replicate the tumor microenvironment than traditional 2D cell cultures, improving drug development and personalized therapy. In this study, we bioprinted gelatin-methacrylol (GelMA)-based organoids containing bladder cancer cell lines of different grades to model muscle-invasive bladder cancer. In the organoids, we observed distinct grade-dependent tumor proliferation and progression dynamics. Treatment with standard-of-care chemotherapies revealed a grade-dependent tumor response consistent with in vivo patient data, highlighting the suitability of these organoids for rapid, reliable drug testing. Lastly, we used the organoids to test LCI139, a novel small-molecule inhibitor of PI3K, CDK4/6, and CDK9 designed for the treatment of epithelial cancers, underscoring the potential of our model to evaluate the efficacy of newly developed drugs. The ability to quickly biofabricate reproducible bladder cancer organoids that are adaptable to different tumor grades represents a novel strategy to create an in vitro platform with strong potential to predict treatment outcomes of bladder cancer patients. Full article

Background/Objectives: Systemic inflammation and circulating proinflammatory cytokines can impair blood–brain barrier (BBB) integrity. Many in vitro BBB models lack the complexity to fully recapitulate systemic inflammation and its long-term effects on the BBB. This study aimed to develop a hollow-fiber flow-based dynamic in vitro (DIV) BBB model for investigating prolonged proinflammatory responses under physiological flow conditions. Methods: Culture conditions for hCMEC/D3 in a DIV model (Flocel) were optimized by varying serum concentrations over seven weeks. Barrier integrity (transendothelial electrical resistance (TEER), permeability studies), proliferation, metabolism (NMR spectroscopy) and molecular changes (high-throughput qPCR) were assessed. Optimized triple-cultures with hCMEC/D3, human primary astrocytes and pericytes were established. After four weeks of barrier establishment, the triple-cultures were exposed to TNF-α, IL-1β and IFN-γ (0.1 ng/mL or 10 ng/mL each) for two weeks. The inflammatory response was assessed with a multiplex cytokine array. Results: Reduced serum concentration (0.25% FBS) decreased proliferation, promoted aerobic respiration, and altered tight junction and transporter gene expression, accompanied by moderately improved barrier integrity compared with 1% or 5% FBS. In optimized triple-cultures, cytokine exposure induced concentration-dependent secretion of IL-6, IL-8, and MCP-1 and changes in mRNA levels, with minor effects on barrier integrity. Sustained cytokine release over two weeks demonstrated stable induction of inflammatory responses at the BBB. Conclusions: An organotypic DIV model of the human BBB, incorporating hCMEC/D3, human primary astrocytes and pericytes was successfully established. By enabling long-term exposure to physiologically relevant cytokine concentrations under flow conditions, this model may provide a platform to investigate functional and molecular BBB responses in inflammation-driven disease progression. Full article

Background: Medical device-associated infections represent a major component of healthcare-associated infections. Biofilm formation promotes microbial persistence on device surfaces, reduces antimicrobial susceptibility, and contributes to multidrug resistance (MDR), complicating diagnosis and treatment. Materials and Method: This study investigated biofilm production and antimicrobial resistance in microorganisms recovered from 100 indwelling and implantable medical devices, including urinary and venous catheters, urethral stents, catheter tips, and orthopedic or prosthetic materials, collected at a tertiary-care hospital (AOU “San Giovanni di Dio e Ruggi d’Aragona”, Salerno, Italy). Microbiological cultures were performed using direct and enrichment methods. Microbial identification was carried out by MALDI-TOF MS, antimicrobial susceptibility testing by VITEK^® (bioMérieux, Marcy-l'Étoile, France) 2 according to EUCAST criteria, and biofilm production was assessed using the crystal violet tissue culture plate assay. MDR status was defined according to international guidelines. Results: Microbial growth was detected in the majority of analized devices, frequently with polymicrobial contamination. Within the study cohort, coagulase-negative staphylococci (CoNS) were the most frequently recovered microorganisms (20%), followed by Klebsiella pneumoniae (10%), Candida albicans (9%), Staphylococcus aureus (9%), Enterococcus faecalis (8%), and Escherichia coli (8%). A significant association was observed between multidrug resistance and biofilm production, with MDR isolates showing a markedly higher likelihood of being biofilm producers compared with non-MDR isolates (OR 9.50; 95% CI 2.72–42.96; p < 0.005). Biofilm formation also differed significantly among device types (p = 0.028). Conclusions: These findings indicate a high prevalence of biofilm-producing MDR microorganisms among isolated recovered from medical devices in our cohort and highlight a significant association between MDR phenotype and biofilm production. These results provide a microbiological characterization of device-associated isolates that may support future studies on infection dynamics and control strategies. Full article

Stem cells, with their self-renewal and multi-lineage differentiation potential, hold promise for tissue repair and intractable diseases treatment. Yet clinical translation of stem cell therapies has long been hindered by insufficient scalable stem cell manufacturing, stemness loss and functional decline in 2D expansion, and poor post-transplantation cell retention, unregulated fate control. Programmable microcarriers (MCs) paired with 3D dynamic culture offer an emerging strategy to address these bottlenecks and enable stem cell fate regulation. In this review, we systematically review advanced MC fabrication strategies for stem cell fate regulation, comparing features of emerging technologies (microfluidics, electrospraying, in-air microfluidics, integrated in situ functionalization) and their implications for programmable MC control and scalable manufacturing. We analyze how MCs modulate stem cell behaviors (adhesion, proliferation, stemness maintenance, differentiation) via synergistic static physicochemical cues and dynamic stimuli-responsive properties. We map the latest advances in functionalized MC-mediated stem cell therapy across osteochondral defects, autoimmune, skin, ophthalmic and neurodegenerative diseases. Finally, we pinpoint unresolved challenges for clinical translation of MC–stem cell system and outline key future research directions. This review offers a systematic roadmap for advancing programmable MC fabrication, clinical-grade stem cell biomanufacturing, and precise cell therapy development. Full article

Time-domain dynamic full-field optical coherence tomography (D-FFOCT) is a powerful label-free imaging modality that enables functional visualization of cellular activity in living tissues with subcellular resolution. However, its sensitivity remains a major limitation for imaging highly scattering three-dimensional (3D) biological models such as retinal organoids, where incoherent background and inefficient optical flux distribution reduce dynamic contrast and limit imaging depth. In this work, we introduce a ratio-free optical configuration for time-domain D-FFOCT that enables continuous tuning of the sample-to-reference field ratio while minimizing photon losses and suppressing parasitic reflections. This polarization-based architecture allows optimal redistribution of optical flux according to sample scattering conditions and improves sensitivity under both power-limited and dose-limited conditions. Compared with conventional non-polarizing beam splitter configurations, the proposed approach provides a $\sqrt{2}$-fold (3 dB) sensitivity improvement through optical optimization alone. In addition, we investigate for the first time the use of partial-field illumination (PFI) in time-domain D-FFOCT to reduce incoherent background arising from multiple scattering. In retinal organoids imaged at 120 $\mathsf{\mu }$m depth, PFI yields up to a 14.5-fold (23.2 dB) increase in dynamic signal sensitivity, while preserving functional contrast. When combined, ratio-free detection and PFI provide a cumulative sensitivity improvement of 20.5-fold (26.2 dB). These gains enable improved cellular-scale visualization in retinal organoids, including cell-resolved imaging within rosette regions, as well as improved detection of intracellular dynamics in Müller glial cell cultures. This work establishes a practical framework for sensitivity optimization in D-FFOCT and expands its potential for functional imaging, disease modeling, and live-cell monitoring in complex biological systems. Full article

Scientific theories survive on institutional fitness, not empirical merit alone. Under Soviet Stalinism, Vygotsky and Luria’s cultural-historical psychology was suppressed while Leontiev’s Activity Theory flourished because it aligned with Marxist-Pavlovian materialism. A game-theoretic framework formalizes this dynamic through three coupled mechanisms: a researcher utility function (U_r = αT + βR − γC), a state utility function (U_s(e) = δI(e) − εD(e) − κ(e)), and a replicator dynamic for institutional selection. Under sufficiently high punishment coefficients, the unique Nash equilibrium is aligned with the ideologically safe theory regardless of empirical truth, and the replicator dynamics drive empirically stronger theories to extinction in the institutional population. Classical findings on conformity and obedience from Sherif, Asch, Festinger, Schachter, and Milgram supply the foundations for the model’s parameters. This pattern—termed here as epistemological selection pressure—explains the Vygotsky case. Because the model assumes severe punishment, active enforcement, complete information, and a binary choice, it applies most directly to authoritarian science; contemporary liberal institutions correspond to the low-punishment regime in which the same model predicts that empirical merit can prevail, so the mechanism is expected to recur only in attenuated form within specific high-pressure domains where scientific truth and institutional power remain entangled. Full article

In this narrative review, the systematic tailoring of cell-derived secretomes for regenerative medicine is comprehensively discussed. The review begins by evaluating a diverse array of cell sources, including platelets, endothelial cells, osteoblasts, chondrocytes, tenocytes, and mesenchymal stem cells (MSCs). From this foundation, we critically analyze how modulating cell culture conditions can be used to directly influence the therapeutic and paracrine profiles of the resulting secretomes. Specifically, we address the impact of the following critical parameters on secretome fabrication: (i) Culture Medium Supplementation: The use of defined media, supplements, and chemical priming agents. (ii) Cell Cultivation Formats: A comparative analysis of 2D monolayer cultures versus 3D spheroids. (iii) Culture Duration and Fluidics: The effects of culture time alongside static versus dynamic culture systems. (iv) Microenvironmental Stress: The influence of varying oxygen levels (for example, hypoxia) and induced oxidative stress on secretory output. Full article

The extracellular matrix (ECM) regulates skeletal muscle development through biochemical signaling and mechanical interactions. While Matrigel supplementation is commonly used to enhance engineered muscle formation, the contribution of specific ECM proteins remain incompletely defined in 3D systems. Here, we evaluated the effects of laminin and fibronectin supplementation on myogenic differentiation in collagen type I hydrogels and assessed their influence on passive tissue compaction and alignment in 3D constructs. Two-dimensional collagen hydrogels supplemented with increasing concentrations (0–100 µg/mL) of laminin or fibronectin were screened to maximize the myoblast fusion index. These concentrations were incorporated into 3D myocyte-seeded hydrogels cultured between flexible posts to quantify passive compaction forces via cantilever mechanics. Fibronectin supplementation (10 µg/mL) resulted in significantly greater early post displacement and sustained passive compaction compared to laminin-supplemented and unsupplemented controls. Constructs cultured under tension between posts exhibited enhanced alignment, with fibronectin further increasing the proportion of fibers oriented within 0–20° of the tension axis. Together, these findings demonstrate that fibronectin enhances early passive compaction dynamics and tension-mediated alignment in collagen-based skeletal muscle constructs. These results provide insight into how specific ECM components influence 3D tissue organization and may inform the design of engineered muscle models for regenerative applications. Full article