Search Results (1,113)

Insect hemocytic cell lines offer substantial advantages over primary, in vivo hemocyte cultures, fundamentally transforming experimental approaches in cellular immunology and related fields. Selected Malacosoma disstria cell lines were characterized for optimal growth temperatures, morphogenesis, blebbing, extracellular enzyme profiles, and their interactions with material (polystyrene) and microbial (Bacillus subtilis) surfaces. The adhesive hemocyte lines UA-Md221 and Md108 showed optimal growth at 28 °C, whereas UA-Md203 and Md66 grew best at 21 °C, with Md66 tolerating 21–28 °C. Md108 demonstrated a broader temperature tolerance than other adherent cultures. Both Md108 and UA-Md221 adhered to polystyrene within 24 h post-subculturing, although protease-induced morphological changes in modified Grace’s medium continued through 48 h and 72 h, respectively. Culture quality was monitored by assessing the release of multiple enzymes, including alkaline and acid phosphatases, esterases and lipases, aminopeptidases, proteases, glycosidases, and hydrolases from the cell lines at 50% confluency in modified Grace’s medium. Fetal bovine serum showed elevated esterase lipase (C8) and phosphoamidase activities when diluted in Grace’s medium and phosphate buffered saline (PBS). Exposure to dead B. subtilis suspended in PBS induced quantitative and qualitative alterations in the enzyme secretion profiles of Md66 and Md108 cultures. We conclude that semi-quantitative assessments of hemocytic cell lines can provide valuable insights for the time window of each enzyme release, revealing immune and metabolic signaling patterns. Full article

Background/Objectives: Postoperative atrial fibrillation (POAF) is a common and serious complication after coronary artery bypass grafting (CABG), leading to increased morbidity and healthcare utilization. Although systemic inflammation is a well-established driver of POAF pathogenesis, no composite preoperative inflammatory biomarker has been validated for risk stratification in this population. This study aimed to evaluate the novel Inflammatory Burden Index (IBI)—the first composite biomarker combining acute-phase (C-reactive protein, CRP) and chronic cellular (neutrophil-to-lymphocyte ratio, NLR) inflammation—as a preoperative predictor of POAF after CABG. Methods: In this large retrospective cohort study, we included 3481 consecutive patients who underwent isolated CABG at a high-volume cardiac center between 2019 and 2024. Preoperative IBI was calculated as CRP (mg/dL) × NLR. The primary outcome was new-onset POAF within the first 7 postoperative days, confirmed by continuous telemetry on 12-lead ECG. Predictive performance was assessed using multivariable logistic regression, receiver operating characteristic (ROC) curve analysis (area under the curve, AUC), net reclassification improvement (NRI), integrated discrimination improvement (IDI), and internal validation via bootstrapping (1000 resamples). Results: POAF developed in 866 patients (24.9%). Patients with POAF exhibited significantly higher preoperative IBI levels (39.4 ± 18.6 vs. 26.3 ± 16.7, p < 0.01). In multivariable analysis adjusted for age, hypertension, left atrial diameter, and other clinical covariates, IBI emerged as a strong independent predictor of POAF (adjusted OR 1.041, 95% CI 1.036-1.046, p < 0.01). The IBI alone demonstrated moderate-to-good discriminative performance (AUC 0.72, 95% CI 0.70–0.74), significantly outperforming the Systemic Immune/Inflammation Index (SII; AUC 0.61, DeLong test p < 0.001) and providing superior reclassification (NRI 0.150, IDI 0.032) and model fit (lower AIC). Combining IBI with established clinical risk factors further improved predictive accuracy (combined AUC 0.74, specificity 72.4%). Tertile-based stratification revealed a clear graded relationship with POAF incidence (low IBI: 16.6%, medium: 21.3%, high: 35.1%; p = 0.02). Notably, the medium IBI stratum (11.18-25.44) displayed the highest discriminative power (AUC 0.87, 95% CI 0.85-0.88), with bootstrap validation confirming model stability (minimal bias, robust 95% CI). Conclusions: This study establishes the preoperative Inflammatory Burden Index (IBI) as the first validated composite inflammatory biomarker independently associated with POAF following CABG. Its superior performance over existing indices (SII), graded risk stratification, and peak accuracy in the moderate inflammation window highlight its potential for personalized preoperative risk assessment and targeted perioperative intervention strategies. Full article

Background: Dry eye disease (DED) is characterized by tear film instability and a hyperosmolar ocular surface, which significantly impacts ocular health. Artificial tear solutions (ATSs) have been effective frontline treatments for DED, yet current commercially available products often provide only temporary relief, necessitating frequent daily administration. Significant efforts have been made to develop next-generation ATSs that can provide prolonged protective effects for DED. High-molecular-weight sodium hyaluronate (HA) is more commonly used in multi-dose preservative ATSs due to its longer chain lengths and rheological properties that can provide an enhanced retention time and clinical comfort and effects. The current methods to evaluate ATSs have largely focused on human biocompatibility and rheological testing and often overlook the dynamic nature of cellular phenotypes or the protective mechanisms at a cellular level. Therefore, this study developed novel in vitro mammalian cell assays involving human corneal epithelial cells (HCECs) to comprehensively assess ATSs with HA for biocompatibility and efficacy. Methods: We evaluated cellular viability across varying severities in two distinct DED models: desiccation and hyperosmotic stress. Simultaneously, time-lapse imaging coupled with computational image analyses quantified subtle, yet significant, cellular morphological changes under these stress condition. Results: Our assays revealed that ATSs provide significant, yet varying, protection against mild, medium, and harsh desiccation stress, as well as hyperosmotic conditions. This study also made a key insight that was the observation that DED conditions induce drastic HCEC morphological changes, including significant cellular monolayer breakage, which were effectively mitigated by the ATS products used in this work. Conclusions: The assays presented here provide a robust standard for ATS testing, ultimately guiding the selection of more effective next-generation therapies and aiding in a greater understanding of DED pathogenesis. Full article

Extracellular vesicles (EVs) are membrane-bound nanoparticles released by almost all cell types into the extracellular space, acting as important mediators of intercellular communication by transferring proteins, lipids, and nucleic acids horizontally. EVs are generally classified into small EVs (<200 nm), medium/large EVs (>200 nm), microvesicles, and apoptotic bodies, with current classification methods focusing on physical properties, molecular composition, and cellular origin, as detailed in the MISEV2023 guidelines. EVs are highly promising for diagnostic and therapeutic applications due to their intrinsic biocompatibility, stability in biological fluids, capacity to carry diverse molecular cargo, and potential for drug delivery and functionalization to enable targeted delivery and tissue repair. This narrative review discusses the emerging roles of EVs across various medical fields, including obstetrics and gynecology, ophthalmology, otorhinolaryngology, urology, oncology, orthopedics, neurology, immunology, wound healing, chronic pain management, dermatology, and cardiology. In each discipline, EVs show potential as biomarkers for diagnosing physiological or pathological conditions and as carriers for targeted drug delivery and regenerative treatments. Exosomes, a major type of small EVs, have especially attracted attention as versatile nanocarriers for precision medicine. However, translation into clinical practice requires addressing key pitfalls, including the standardization of isolation and characterization protocols, dose definition, GMP-compliant large-scale production, and regulatory approval. Ongoing interdisciplinary collaboration across disciplines and thorough clinical testing will be essential to unlock the full biomedical potential of EVs and establish them as transformative tools in personalized healthcare. Full article

A CoCrNi-AlTi medium-entropy alloy was fabricated via laser powder bed fusion (L-PBF), and its microstructural evolution and mechanical response during aging at 500–900 °C for 1 h were systematically investigated. The as-built alloy exhibits a hierarchical microstructure consisting of elongated columnar grains and dislocation-rich cellular substructures, which is associated with an excellent strength–ductility combination (YS: 848 MPa, UTS: 1136 MPa, EF: 32.6%). Upon aging, a pronounced precipitation-hardening response is observed, with a peak hardness of 501 ± 7 HV and an ultimate tensile strength of 1429 MPa achieved at 800 °C. TEM and STEM-EDS analyses indicate that Ti preferentially segregates along dislocation networks and grain boundaries at early aging stages, promoting the heterogeneous nucleation of nanoscale Ni–Al–Ti–rich precipitates that effectively impede dislocation motion. At elevated aging temperatures, additional Cr-enriched regions with diffuse compositional partitioning are observed within the FCC matrix, occurring concurrently with the peak mechanical performance. Further aging at 900 °C leads to strength degradation, which is attributed to precipitate coarsening and recovery-induced dislocation annihilation. These results highlight the critical role of L-PBF-induced defect structures in governing precipitation behavior and the resulting strength–ductility trade-off during post-build heat treatment of CoCrNi-AlTi medium-entropy alloys. Full article

The toxicity of micro- and nanoplastics in aquatic life is well documented, yet limited information is available on their effects in humans; moreover, most in vitro nanotoxicology studies rely on cancer cells. This study examined the effects of pristine and aged polystyrene micro- and nanoparticles on human dental pulp stem cells. While both particle sizes were internalized by the cells, primarily through endocytosis, they did not affect cell viability. In contrast, leachates from particles, aged for one month in culture medium, significantly reduced viability, indicating that toxicity arises from degradation byproducts rather than the particles themselves. Atomic force microscopy confirmed surface changes in aged plastics. Both particle sizes disorganized the cytoskeleton, leading to reduced actomyosin cortex integrity. Gene expression analysis revealed that leachates and aged particles activated inflammatory pathways, markedly increasing IL-8 and TGF-β1 expression, while also decreasing SOD levels associated with oxidative stress. No notable effects were observed on genes related to stemness or senescence. These results suggest that, while pristine micro- and nanoplastics may be relatively inert, their degradation products pose greater toxicological risks to human health. The findings highlight the importance of considering leachate toxicity in plastic pollution studies and demonstrate the value of stem cell-based models for evaluating the cellular and molecular impacts of environmental contaminants on human health. Full article

Bioreactors used for the maturation of cell-seeded tissue-engineered scaffolds should essentially mimic the dynamic in vivo environments experienced by the native tissues they intend to substitute. In addition to perfusion of growth medium to facilitate continuous mass transfer, application of appropriate mechanical stimulation is important to enhance cellular responses in scaffolds for tissues such as tendons, skin, and cardiac muscle that experience dynamic loading. This study focuses on the development of a multi-modal custom bioreactor capable of applying cyclic tensile stimulation and perfusion within physiologically relevant ranges while minimizing shear stress detrimental to cells seeded on scaffolds. To validate the bioreactor design and operation, we assessed the effects of tensile stimulation (0.1 Hz, 2000 cycles/day) and perfusion (media flow rate = 0.15 mL/min) over 21 days on the biofunctional performance of 3D-bioplotted polycaprolactone (PCL) auxetic scaffolds with a representative design (missing-rib pattern) characterized by negative Poisson’s ratio similar to the aforementioned soft tissues. The scaffold had a tensile yield strain of 9.14%, yield strength of 0.25 MPa, elastic modulus of 2.85 MPa, and ultimate tensile strength (UTS) of 1.32 MPa. The application of perfusion and tensile stimulation (0–5% cyclic strain) for 21 days did not adversely affect the yield strength and elastic modulus of the scaffold but affected its UTS (22.5% decrease) compared to the control cultured without perfusion or stimulation. Notably, it resulted in significantly improved fibroblast cellular responses (DNA = 29 µg/g sample and collagen = 371.78 µg/g sample) compared to the control (7.52 µg/g sample and 163.51 µg/g sample, respectively). These results validate the bioreactor system operation and the ability of multi-modal stimulation to control biofunctional responses of auxetic scaffolds, which will serve as the basis for future studies that will optimize auxetic scaffold design and dynamic culture parameters for NPR tissue-specific applications. Full article

Retroviruses are immunosuppressive, and there is evidence that a highly conserved immunosuppressive domain (isu domain) in their transmembrane envelope protein contributes to this activity. Studies have shown that inactivated retroviruses, their purified transmembrane envelope proteins, and synthetic peptides corresponding to the isu domain inhibit mitogen-triggered proliferation of peripheral blood mononuclear cells (PBMCs) and modulate their cytokine and gene expression. This has been demonstrated for human immunodeficiency virus type 1 (HIV-1), as well as for beta- and gammaretroviruses and for both exogenous and endogenous retroviruses, including syncytins. In the case of HIV-1, homopolymers of its isu peptide stimulated an increased release of IL-10, IL-6, and other cytokines from human PBMCs. Up-regulated genes included IL-6, IL-8, and IL-10, as well as MMP-1, TREM-1, and IL-1β. In vivo, in a mouse tumor model, tumor cells that were unable to induce tumors in immunocompetent animals gained the ability to do so when expressing the transmembrane envelope protein or the isu domain of various retroviruses on their surface. Here, we demonstrate that the transmembrane envelope protein p15E of PERV can modulate cytokine expression in human PBMCs. Human 293 cells were transfected with four constructs that express a portion of p15E, including the isu domain, and were cultured in the presence of a selection medium containing hygromycin. The p15E-expressing cells were co-cultured with human PBMCs, leading to the release of IL-6 and IL-10 protein and the modulation of multiple cytokines and other markers, including IL-6, IL-10, IFN-α, TNF-α, MMP1, and SEPP1. Similar, but more pronounced, effects were observed when PERV-producing 293 and pig cells were used in parallel; both expressed higher levels of p15E. Additionally, p15E expression reduced MHC class I expression, and preliminary data indicate that p15E expression could have a protective effect against cellular cytotoxicity. This finding underscores the need for further research to elucidate the dynamics of p15E expression and its immunosuppressive activity. It also contributes to the understanding of the immunosuppressive properties of pathogenic retroviruses. Furthermore, expressing the immunosuppressive p15E of PERV on the surface of a pig xenotransplant may reduce the need for pharmaceutical immunosuppressants. Full article

The creation of a specific culture medium for colorectal organoids in 2011 heralded a new era in human primary cultures by enabling the indefinite expansion of normal and pathological epithelial organoids. The original formula has been used ever since, with only minor, lab-specific modifications. The goal of culturing organoids from different tissues has relied on saving and propagating the pluripotent stem cell. The “magic bullet” and all its subsequent derivatives have pursued this goal. Consequently, agonist and antagonist signals are chronically activated in the organoid medium, forcing organoid cells (as well as any other co-cultured cellular model) into constrained signaling pathways. This extremely artificial condition is often overlooked in experimental approaches and may bias the results. Furthermore, some molecules in the organoid medium have unpredictable off-target effects that significantly impact the behavior and maturation of certain cell populations. Nicotinamide, gastrin and PGE2 inhibit immune responses. SB202190, A83-01 and vanadate (from advanced DMEM-F12) modify intracellular signaling. N-AcetylCysteine and Primocin modify the redox response and mitochondrial metabolism, respectively. Thus, the unintentional addition of these molecules to the organoid medium introduces biases under specific experimental settings. While the original organoid medium formula is the gold standard for propagating organoids in vitro, more focused, reliable conditions are necessary for specific organoid-based tests. Full article

Glucocorticoids are widely applied intra-articularly to alleviate inflammation and pain in osteoarthritis (OA). However, repeated administration and high local concentrations can lead to crystal deposition on the cartilage surface, contributing to chondrocyte damage and extracellular matrix (ECM) degradation, potentially accelerating OA progression. Calcium-dependent mechanosensors play a critical role in mediating catabolic responses in chondrocytes, but it remains unclear whether glucocorticoids affect chondrocyte mechanosensitivity or biomechanical properties. This in vitro study examined the dose-dependent effects of triamcinolone acetonide (TA) on chondrocyte biomechanics and mechanosensitivity. Primary human chondrocytes (N = 23) were cultured for one week with TA (2 µM–2 mM) or control medium. Cytoskeletal organization was visualized by F-actin staining (N = 6), and cellular elasticity (N = 5) was quantified via atomic force microscopy (AFM). Mechanotransduction was analyzed by Ca²⁺ imaging (Fluo-4 AM) upon AFM-based indentation (500 nN). Expression of matrix-related and mechanosensitive genes (N = 9) was assessed by qPCR. TA exposure induced a concentration-dependent reorganization of the F-actin cytoskeleton, pronounced at 0.2 mM, accompanied by a significant increase in the elastic modulus (p < 0.001). TA further augmented Ca²⁺ fluorescence intensity under basal conditions and during mechanical stimulation. Blocking cationic mechanosensitive channels with GsMtx4 (N = 3) markedly reduced the TA-evoked Ca²⁺ influx (p < 0.0001). Significant reduction in MMP1 was observed on the transcriptional level (N = 9) after TA-treatment (p < 0.05). In summary, TA enhances chondrocyte stiffness through cytoskeletal condensation and amplifies Ca²⁺-dependent mechanotransduction but reduces MMP1 expression, indicating a dual biomechanical response of chondrocytes to OA under exposure of potent corticosteroid. Full article

Acetyl phosphate (AcP) is a microbial metabolite acting as a link between cell metabolism and signaling, providing the survival of bacteria in the host. AcP was also identified as an intermediate of pyruvate oxidation in mammalian mitochondria and was found in the human blood in some severe pathologies. The possible contribution of circulating AcP to the maintenance of the physiological or pathological states of the body has not been studied. Since AcP can function as a donor of phosphate groups, we have examined in vitro the influence of AcP on calcium signaling in mitochondria and cells by measuring the membrane potential and the calcium retention capacity of mitochondria by selective electrodes and by assaying the cell calcium signaling by Fura-2AM fluorescent radiometry. AcP was shown to induce a concentration-dependent increase in the mitochondrial resistance to calcium ion loading both in the control and in the presence of ADP. This effect was especially pronounced when mitochondria were incubated in a phosphate-free medium; under these conditions, AcP strongly raised the membrane potential and increased the rate of calcium uptake and the calcium retention capacity several times. Moreover, AcP induced similar changes in human cells when calcium signaling was activated by ATP, to a greater extent in neuroblastoma cells than in astrocytes. In the presence of AcP, a tendency for an increase in the amplitude and a decrease in the continuance of the ATP-induced calcium response was observed. These changes are probably associated with the activation of calcium buffering by mitochondria due to the delivery of phosphate during the hydrolysis of AcP. The results show that AcP is involved in the regulation of the Ca²⁺ balance in cells by activating the accumulation of calcium ions by mitochondria, especially under phosphate deficiency. A shift in calcium signaling mediated by AcP supplementation may be caused by hyperphosphatemia, which is now considered as one of basic contributors to cellular dysfunction and progression of various diseases, including sepsis. Full article

Anthocyanin is a flavonoid known for its strong antioxidant and anti-inflammatory activities in both in vitro and in vivo systems. This study investigated whether anthocyanin supplementation could improve the developmental competence, hatching rate, and the expression of development- and proliferation-related markers in ICR mouse blastocysts cultured in vitro. Mouse embryos were cultured in KSOM medium supplemented with 2, 4, or 8 μM anthocyanin. Among these, 4 μM was selected as the working concentration within the tested range. Morphological assessment was used to evaluate blastocyst development and hatching, while quantitative real-time polymerase chain reaction (qPCR) was performed to measure the expression of GLUT4 and PI3K. Anthocyanin supplementation significantly enhanced blastocyst quality, as reflected by higher developmental competence and increased hatching rates compared with the control group. In addition, anthocyanin-treated blastocysts displayed elevated mRNA expression of GLUT4 and PI3K, indicating a potential association with enhanced metabolic readiness and cellular proliferation. Overall, these findings indicate that anthocyanin supports embryo quality during preimplantation development in vitro, with potential relevance to implantation-related processes. Further research is needed to clarify the underlying mechanisms and explore the potential applications of anthocyanin in reproductive medicine. Full article

Microbial contamination of food is a crucial cause of food spoilage and foodborne diseases, posing a severe threat to global public health. Although chemical preservatives are effective, their potential hazards to human health and the environment, coupled with the growing demand for “clean label” products, have driven the search for natural alternatives. Lactic acid bacteria (LAB), recognized as the Generally Recognized as Safe (GRAS) microorganisms, have emerged as the promising bio-preservatives due to their safety, effectiveness, and multifunctionality. This review systematically summarized the core antimicrobial properties of LAB, including their inhibitory spectrum against foodborne pathogens, spoilage microorganisms, viruses, parasites, and their ability to degrade toxic substances such as mycotoxins, pesticides, and heavy metals. Key inhibitory mechanisms of LAB are highlighted, encompassing the production of antimicrobial metabolites, leading to metabolism disruption and cell membrane damage, nutrition and niche competition, quorum-sensing interference, and anti-biofilm formation. Furthermore, recent advances in LAB applications in preserving various food matrices (meat, dairy products, fruits and vegetables, cereals) are integrated, including their roles in enhancing food sensory quality, extending shelf life, and retaining nutritional value. The review also discusses critical factors influencing LAB’s inhibitory activity (medium composition, culture conditions, ionic components, pathway regulator, etc.) and the challenges associated with the application of LAB. Finally, future research directions are outlined, including the novel LAB and metabolites exploration, AI-driven cultural condition optimization, genetic engineering application, nano-encapsulation and active packaging development, and building up the LAB-based cellular factories. In conclusion, LAB and their antimicrobial metabolites hold great promise as green and safe food preservatives. This review is to provide comprehensive theoretical support for the rational improvement and efficient application of LAB-based natural food preservatives, contributing to the development of a safer and more sustainable food processing and preservation systems. Full article

Metamaterials (MMs)-based terahertz (THz) biosensors hold promise for clinical diagnosis, featuring label-free operation, simple, rapid detection, low cost, and multi-cell-type discrimination. However, liquid around cells causes severe interference to sensitive detection. Most existing MMs-based cell biosensors detect dead cells without culture medium (losing original morphology), hindering stable, sensitive multi-cell discrimination. Here, a terahertz biosensor composed of a microcavity and MMs can be used to detect and discriminate multiple cell types within suspension. Its detection mechanism relies on cellular size (radius)/density in suspension, which induces effective permittivity (ε_eff) differences. By designing MMs’ split rings with luxuriant gaps, the biosensor achieves a theoretical sensitivity of ~328 GHz/RIU, enabling sensitive responses to suspended cells. It shows a robust, increasing frequency shift (610–660 GHz) over 72 h of cell apoptosis. Moreover, it discriminates nerve cells, glioblastoma (GBM) cells, and their 1:1 mixture with obviously distinct frequency responses (~650, ~630, ~620 GHz), which suggests effective and reliable multi-cell-type recognition. Overall, this study and its measurement method should pave the way for metamaterial-based terahertz biosensors for living cell detection and discrimination, and this technology may inspire further innovations in tumor investigation and treatment. Full article

The FDA plans to gradually replace animal testing with organoid and organ-on-a-chip technologies for drug safety assessment, driving surging demand for gut-on-a-chip in food and drug safety evaluation and highlighting the need for efficient, precise chip designs. Oxygen gradients are central to these devices because they shape epithelial metabolism, microbial co-culture, and overall gut homeostasis. We coupled machine learning with finite element analysis to build a parametric COMSOL Multiphysics model linking channel geometry, transport coefficients, and cellular oxygen uptake to the resulting oxygen field. For numerical prediction, three models—Random Forest (RF), XGBoost, and MLP—were employed, with XGBoost achieving the highest accuracy (RMSE = 1.68%). SHAP analysis revealed that medium flow rate (39.7%), external flux (26.9%), and cellular oxygen consumption rate (24.8%) contributed most importantly to the prediction. For oxygen distribution mapping, an innovative Boundary-Guided Generative Network (BG-Net) model was employed, yielding an average concentration error of 0.012 mol/m³ (~4.8%), PSNR of 33.71 dB, and SSIM of 0.9220, demonstrating excellent image quality. Ablation experiment verified the necessity of each architectural component of BG-Net. This pipeline offers quantitative, data-driven guidance for tuning oxygen gradients in gut-on-a-chip. Future work will explore extensions including real experimental data integration, real-time prediction, and multi-task scenarios. Full article