Search Results (4,182)

Human mesenchymal stem cells (hMSCs) and their secreted extracellular vesicles (EVs) are promising therapeutics to treat degenerative or inflammatory diseases such as ischemic stroke and Alzheimer’s disease (AD). hMSC-EVs have the coveted ability to contain therapeutically relevant biomaterials; however, EV biogenesis is sensitive to the culture microenvironment in vitro. Recently, the demand for hMSC-EVs has increased dramatically, highlighting the need for scalable bioreactors for large-scale biomanufacturing. In this study, adipose-derived hMSCs were seeded in 2D plates, an ultralow-attachment (ULA) plates as static aggregates, a novel vertical wheel bioreactor (VWBR) as aggregates, and a spinner flask bioreactor (SFB). EV secretion was quantified and compared using ExtraPEG-based ultracentrifugation and nanoparticle tracking analysis. Compared to the 2D group, significantly higher total EV production and cell productivity in the bioreactors were observed, as well as the upregulation of EV biogenesis genes. Furthermore, there was increased EV production in the VWBR compared to the SFB and the static ULA control. Functional assessments demonstrated that EVs, when delivered via culture medium or hydrogel-based systems, significantly attenuated oxidative stress elevation, suppressed proinflammatory cytokine secretion (e.g., TNF-α) and gene expression, and inhibited nuclear factor kappa-light-chain-enhancer of activated B-cell (NF-κB) activation and neurodegenerative markers across in vitro assays. These findings suggest EV-mediated mitigation of oxidative and inflammatory pathways, potentially through modulation of the NF-κB signaling cascade. This study shows the influence of bioreactor types and their microenvironments on EV secretion in hMSCs and their applications in hMSC-EV production and bioengineering. Full article

Thermoresponsive hydrogels that undergo reversible sol-gel transitions near physiological temperatures are highly attractive for biomedical applications, such as injectable drug delivery and embolization therapies. In this study, a library of polyurethane-based hydrogels was synthesized via step-growth polymerization using polyethylene glycol (PEG) of varying molecular weights, different diisocyanates, and a series of functional diols derived from diethanolamine with increasing hydrophobicity. The resulting polymers exhibited sol–gel transition behaviors without the need for external crosslinkers, relying solely on non-covalent interactions. The thermal responsiveness was systematically investigated using UV–Vis turbidimetry, and the cloud point temperature (T_CP) was found to be tunable within a range of 26–49 °C by modulating the monomer composition. Statistical modeling identified PEG molecular weight and diol structure as the primary determinants of T_CP, while diisocyanate type and diol-to-PEG ratio had negligible effects. Only diethanolamine (DEA)-based polymers formed stable hydrogels above a critical gelation temperature (LCGT), attributed to enhanced intermolecular interactions via free amine groups. In vitro degradation assays confirmed good hydrolytic stability under physiological conditions over four weeks, with degradation profiles strongly influenced by the PEG chain length and hydrophobic content. These findings establish a structure–property framework for the rational design of injectable, thermoresponsive polyurethane hydrogels with tailored sol–gel behavior for biomedical applications. Full article

Cowpea (Vigna unguiculata L. Walp.) is a species with superior tolerance to drought stress compared to other legumes. It is a promising crop with increasing importance in the face of global climate changes. Local forms, well adapted to particular agro-climatic conditions, are useful germplasm resources. Five Bulgarian cowpea landraces, which had displayed differences in osmotic stress tolerance at the germination stage, were subjected to severe stress (15% PEG 6000 in Hoagland nutrient media) during 16 days at the vegetative growth stage (plants with expanded trifoliate leaves). All local forms responded to the imposed stress by biomass and leaf area diminution, a slight increase in leaf water deficit and electrolyte leakage, proline accumulation in roots and leaves, and an increase in root starch and leaf phenol content. Roots presented more pronounced metabolic changes than leaves, including increased total antioxidant activity, phenolic and carbohydrate content, and proline accumulation. Under osmotic stress, tight control of oxidative stress and concerted upregulation of superoxide dismutase, catalase, glutathione transferase, and peroxidase activities in leaves were registered along with changes in certain specific isoforms, while glutathione reductase activity diminished. Antioxidant enzyme activities had different changes in stressed roots, compared to leaves, and among genotypes. The accession most sensitive to osmotic stress at germination presented more symptoms of oxidative stress at the vegetative growth stage. Full article

Advancements in bioinks and three-dimensional (3D) printing and bioprinting have significantly advanced cardiovascular tissue engineering by enabling the fabrication of biomimetic cardiac and vascular constructs. Traditional 3D printing has contributed to the development of acellular scaffolds, vascular grafts, and patient-specific cardiovascular models that support surgical planning and biomedical applications. In contrast, 3D bioprinting has emerged as a transformative biofabrication technology that allows for the spatially controlled deposition of living cells and biomaterials to construct functional tissues in vitro. Bioinks—derived from natural biomaterials such as collagen and decellularized matrix, synthetic polymers such as polyethylene glycol (PEG) and polycaprolactone (PCL), or hybrid combinations—have been engineered to replicate extracellular environments while offering tunable mechanical properties. These formulations ensure biocompatibility, appropriate mechanical strength, and high printing fidelity, thereby maintaining cell viability, structural integrity, and precise architectural resolution in the printed constructs. Advanced bioprinting modalities, including extrusion-based bioprinting (such as the FRESH technique), droplet/inkjet bioprinting, digital light processing (DLP), two-photon polymerization (TPP), and melt electrowriting (MEW), enable the fabrication of complex cardiovascular structures such as vascular patches, ventricle-like heart pumps, and perfusable vascular networks, demonstrating the feasibility of constructing functional cardiac tissues in vitro. This review highlights the respective strengths of these technologies—for example, extrusion’s ability to print high-cell-density bioinks and MEW’s ultrafine fiber resolution—as well as their limitations, including shear-induced cell stress in extrusion and limited throughput in TPP. The integration of optimized bioink formulations with appropriate printing and bioprinting platforms has significantly enhanced the replication of native cardiac and vascular architectures, thereby advancing the functional maturation of engineered cardiovascular constructs. Full article

Catalpa bungei C.A.Mey is an economically significant deciduous tree valued for timber production and landscaping applications. An efficient regeneration system is crucial for clonal propagation and serves as a foundation for future molecular breeding in C. bungei. This study established two in vitro regeneration pathways—indirect somatic embryogenesis and shoot organogenesis utilizing mature zygotic embryos as explants. Primary callus was induced from cotyledon, hypocotyl, and plumule explants. A high frequency (45.73%) of yellow-green compact callus was achieved on De-Klerk and Walton (DKW) medium supplemented with 2.0 mg/L 6-BA, 1.0 mg/L zeatin (ZT), and 0.1 mg/L NAA. Subsequent transfer to 1.5× Murashige and Skoog (MS) medium containing 1.5 mg/L 6-BA, 0.2 mg/L ZT, and 0.1 mg/L NAA yielded the highest embryogenic callus induction rate (16.67%). Embryogenic callus demonstrated bipotent potential, generating both adventitious shoots and somatic embryos under specific hormonal conditions. Histological analyses confirmed the typical developmental stages of somatic embryos, from globular to cotyledonary forms, validating the embryogenic origin of regenerated structures. Furthermore, hormone or osmotic additives such as abscisic acid (ABA), Phytagel, and polyethylene glycol 4000 (PEG4000) significantly enhanced somatic embryo induction, with Phytagel at 5.0 g/L achieving the highest rate (76.31%). For shoot organogenesis, the optimal hormonal combination of the 0.6 mg/L 6-BA, 0.4 mg/L KT, and 0.15 mg/L NAA achieved the highest bud induction rate (88.89%) and produced an average of 4.07 adventitious buds per explant. This study presents an efficient regeneration system for C. bungei, providing a practical platform for large-scale propagation and basis for biotechnological applications in woody plants. Full article

Natural polymers, such as gelatin, offer a sustainable, green, and versatile alternative for developing proton exchange membranes in low-temperature fuel cell applications. They provide a balance of biocompatibility, flexibility, and ionic conductivity. In this work, gelatin-based composite membranes are reported. The membranes were fabricated and modified with various additives, including ionic liquids (ILs), polyethylene glycol (PEG), and glycerol, to enhance their electrochemical and mechanical properties. The proton conductivity of the pure gelatin membrane was relatively low at 1.0368 × 10⁻⁴ Scm⁻¹; however, the incorporation of IL ([DEMA][OMs]) significantly improved it, with the gelatin/0.2 g IL membrane achieving the highest conductivity of 4.181 × 10⁻⁴ Scm⁻¹. The introduction of PEG and glycerol also contributed to enhanced conductivity and flexibility. The water uptake analysis revealed that IL-containing membranes exhibited superior hydration properties, with the highest water uptake recorded for the gelatin/0.2 g glycerol/0.2 g IL membrane, which was found to be very high (906.55%). The results showed that the combination of IL and PEG provided enhanced proton transport and mechanical stability (as examined visually), making these membranes promising candidates for fuel cell applications. Therefore, this study underscores the importance of bio-based materials by utilizing gelatin as a sustainable, biodegradable polymer, supporting the transition toward greener energy materials. The findings demonstrate that modifying gelatin with conductivity-enhancing and plasticizing agents can significantly improve its performance, paving the way for bio-based proton exchange membranes with improved efficiency and durability. Full article

Introduction: Prolonged fasting induces histological and ultrastructural changes of the intestinal mucosa that may reduce absorption in malnourished patients with high risk of refeeding syndrome. Endocrine function of the intestinal mucosa may be affected by starvation with potential implications for nutritional support. Objective: The present study aims to evaluate the expression of gastrointestinal hormones in duodenal enteroendocrine cells (EECs) of patients after a long starvation period and to assess the changes in EEC hormonal expression after enteral refeeding in the same individuals. Methods: This was an observational prospective controlled study. Adult patients submitted to endoscopic gastrostomy (PEG) with an ingestion below 50% of daily needs for at least one month were enrolled. Duodenal biopsies were collected before gastrostomy (T0) and after 3–6 months of PEG feeding (T1). Biopsies underwent immunohistochemical analysis for chromogranin-A (CgA), neurotensin and incretin (GLP-1 and GIP) tissue expression. Normal duodenum biopsies were used as controls. Results: A total of 30 patients (16 men/14 women) aged 67.1 ± 13.5 years were included, and 14 patients completed follow-up at both periods (46.7%). Malnutrition was diagnosed in all patients according to GLIM criteria. T0 tissue expression defined by median stained area for CgA, GLP-1, and GIP were significantly higher in patients compared to controls (CgA: 1.04% vs. 0.41%; GLP-1: 0.17% vs. 0.03%; GIP: 0.19% vs. 0.03%) (p < 0.001) without differences for neurotensin (0.01%) (p = 0.96). T1 hormonal tissue expression was not significantly reduced after 3–6 months of enteral refeeding (p > 0.05). Conclusions: Prolonged fasting induces increased expression of incretins and chromogranin-A in the duodenum that probably reflect an adaptative response to maintain the anabolic insulin effect under nutritional deficiency. Hormonal expression does not normalize after PEG refeeding during a short period. Full article

Angiotensin-converting enzyme 2 (ACE2) is a key mediator of SARS-CoV-2 host cell entry, making it an attractive target for drug delivery strategies. Nafamostat (NM), a multifunctional agent with antiviral and anti-inflammatory properties, holds promise for COVID-19 treatment. In this study, we developed PLGA-PEG nanoparticles encapsulating NM (NM-PP NPs) and further conjugated them with specific ACE2 decoys (CTC-445.2d or SI5α) to generate NM-PP-Pro/Pep NPs. Both unmodified and ACE2-decoy-modified NPs exhibited uniform size distributions (diameter < 200 nm) and negative surface charges, as confirmed by dynamic light scattering and zeta potential measurements. The nanoparticles maintained structural integrity for at least 18 days at 4 °C and room temperature. In vitro release studies revealed sustained and controlled NM release kinetics. Notably, NM-PP-Pro NPs displayed potent antiviral activity, with an IC₅₀ < 0.05 nM against wild-type SARS-CoV-2 and remained effective against the D614G variant (IC₅₀ = 2 nM). These results underscore the potential of NM-PP-Pro NPs as a versatile n;anotherapeutic platform for targeting SARS-CoV-2 and its emerging variants. Full article

The growing threat of antimicrobial resistance drives the need for innovative and multifunctional therapeutic systems. In this study, a controlled-release system based on a bioactive film composed of gelatin, bacterial cellulose (BC), sericin, citric acid, PEG 400, and nisin was developed for topical applications in infected wound treatment. BC membranes were produced using Komagataeibacter xylinus and enzymatically treated to optimize dispersion within the polymer matrix. The resulting system exhibited a semi-rigid, homogeneous morphology with appropriate visual characteristics for dermatological use. Microbiological assays demonstrated significant antimicrobial activity against Gram-positive (Staphylococcus aureus) and resistant Gram-negative strains (Escherichia coli and Enterobacter cloacae), attributed to the synergistic action of nisin and citric acid, which enhanced bacterial outer membrane permeability. The antioxidant capacity was confirmed through DPPH radical scavenging assays, indicating a progressive release of bioactive compounds over time. Scanning electron microscopy (SEM) analyses revealed good integration of biopolymers within the matrix. These results suggest that the strategic combination of natural biopolymers and antimicrobial agents produced a functional system with improved mechanical properties, a broadened antimicrobial spectrum, and promising potential as a bioactive wound dressing for the treatment of infected skin lesions. Full article

This study proposes a rigid-flexible neural optrode integrated with anti-bending SU-8 optical waveguides and locally soft peptide-functionalized microelectrodes to address the challenges of precise implantation and long-term biocompatibility in traditional neural interfaces. Fabricated via microelectromechanical systems (MEMS) technology, the optrode features a PBK/PPS/(PHE)₂ trilayer electrochemical modification that suppresses photoelectrochemical (PEC) noise by 63% and enhances charge storage capacity by 51 times. A polyethylene glycol (PEG)-enabled temporary rigid layer ensures precise implantation while allowing post-implantation restoration of flexibility and enabling positioning adjustment. In vitro tests demonstrate efficient light transmission through SU-8 waveguides in agar gel and a 63% reduction in PEC noise peaks. Biocompatibility analysis reveals that peptide-coated PI substrates improve cell viability by 32.5–37.1% compared to rigid silicon controls. In vivo validation in crucian carp midbrain successfully records local field potential (LFP) signals (60–80 μV), thereby confirming the optrode’s sensitivity and stability. This design provides a low-damage and high-resolution tool for neural circuit analysis. It also lays a technical foundation for future applications in monitoring neuronal activity and researching neurodegenerative diseases with high spatiotemporal resolution. Full article

SARS-CoV-2 entry into host cells involves multiple steps and is a highly orchestrated process. Both the host protease TMPRSS2 and the HR1/HR2 segment within the spike (S) protein play a crucial role in promoting viral invasion. Herein, we report a series of bifunctional SARS-CoV-2 entry inhibitors formed by covalently linking a TMPRSS2 inhibitor, Camostat (Cm), and an HR1-targeting peptide fusion inhibitor IPB19 via a poly (ethylene glycol) (PEG) linker. Among them, IP4X and IP4Z display potent inhibitory activities against SARS-CoV-2 with similar IC₅₀ values of 0.16 μM and 0.17 μM, respectively. The efficacy surpassed that of their parent inhibitors by approximately 28-fold relative to Camostat and 15-fold relative to IPB19. We confirm that IP4X and IP4Z exhibit a dual-targeting mechanism by binding to both TMPRSS2 and HR1 region of S protein. These findings highlight the potential of the bifunctional inhibitors for further development as a novel multitarget therapy against SARS-CoV-2 infection and enrich the understanding of S-mediated entry of SARS-CoV-2 into host cells. Full article

Welsh onion (Allium fistulosum L.), a globally significant vegetable, flavoring agent, and phytomedicine resource, has remained unavailable with established transient expression platforms for functional genomic investigations. To address this critical methodological limitation, we present systematically optimized protocols for both Agrobacterium-mediated hairy root transformation and protoplast transient expression systems, achieving significant advances in transformation efficiency for this species. Through systematic optimization of key parameters, including Agrobacterium rhizogenes (A. rhizogenes) strain selection (with Ar.Qual demonstrating superior performance), explant type efficacy, bacterial suspension optical density (OD₆₀₀ = 0.3), and acetosyringone induction concentration (100 μM), we established a highly efficient stem disc infection methodology, achieving 88.75% hairy root induction efficiency. Subsequent optimization of protoplast isolation protocols identified the optimal enzymatic digestion conditions: 6-h dark digestion of young leaves using 1.0% (w/v) Cellulase R-10, 0.7% (w/v) Macerozyme R-10, and 0.4 M mannitol, yielding 3.3 × 10⁶ viable protoplasts g⁻¹ FW with 90% viability. System functionality validation through PEG-mediated transient transformation demonstrated successful green fluorescent protein (GFP) reporter gene expression, confirmed by fluorescence microscopy. As the first documented transient expression platforms for Welsh onion, these protocols enable essential molecular investigations, including in planta promoter activity profiling, subcellular protein localization, and CRISPR-based genome-editing validation. This methodological breakthrough overcomes previous technical constraints in Welsh onion molecular biology, providing critical tools for accelerated gene functional characterization in this agriculturally important species. Full article

Heated tobacco products (HTPs) frequently induce user discomfort due to high mainstream smoke temperatures. To address this challenge and improve the inhalation experience, this experiment designed and prepared a cage-shaped braided tube as the cooling section of the filter for HTPs. The thermal, cooling, suction resistance, and smoke composition properties of the filter were tested and analyzed. Thermal analysis (DSC/TG) revealed a 116.53 J/g increase in endothermic enthalpy for PEG-impregnated samples, accompanied by maintained thermal stability (decomposition temperature ≈ approximately 350 °C). The 0.8 wt% Carbon Nanotube (CNT) composite achieved exceptional thermal conductivity (0.597 W/m·K), representing a 521% improvement over untreated controls. The braided tube optimal performance (3 mm inner diameter, 30% PEG/0.8% CNT) reduced the highest smoke temperature to 47.8 °C while maintaining acceptable suction resistance (68.5 Pa, 56.4% reduction vs. commercial IQOS filters). GC-MS analysis confirmed negligible alterations in smoke composition (p > 0.05). This innovation offers an effective thermal management solution that does not compromise sensory experiences. Full article

Drought stress limits seedling growth, hindering morphological development and population establishment. Artocarpus nanchuanensis, a critically endangered species endemic to the karst regions of southwest China, exhibits poor population structure and limited natural regeneration in the wild, with water deficit during the seedling stage identified as a major factor contributing to its endangered status. Elucidating the physiological and molecular mechanisms underlying drought tolerance in A. nanchuanensis seedlings is essential for improving their drought adaptability and facilitating population recovery. In this study, 72 two-year-old seedlings were divided into two groups: drought (PEG) and ethephon (PEG + Ethephon), and subjected to drought-rehydration experiments. The results showed that exogenous application of 100 mg·L⁻¹ ethephon significantly improved stomatal conductance and photosynthetic pigment content in A. nanchuanensis seedlings. Under drought stress, the PEG + Ethephon group exhibited rapid stomatal closure, maintaining water balance and higher photosynthetic pigment levels. After rehydration, the PEG + Ethephon group significantly outperformed the PEG group in terms of photosynthetic rate. Ethephon treatment reduced H₂O₂ and MDA levels, enhanced antioxidant enzyme activity (SOD, CAT, POD, GR), and increased osmotic regulator activity (soluble sugars, soluble proteins, and proline), improving ROS-scavenging capacity and reducing oxidative damage. Ethephon application significantly enhanced ethylene accumulation in seedlings, while drought stress stimulated the concentrations of key ethylene biosynthetic enzymes (SAMS, ACS, and ACO), thereby further contributing to improved drought resistance. Transcriptomic data revealed that drought stress significantly upregulated key ethylene biosynthesis genes, with expression levels increasing with stress duration and rapidly decreasing after rehydration. WGCNA analysis identified eight key drought-resistance genes, providing valuable targets for future research. This study provides the first mechanistic insight into the physiological and molecular responses of A. nanchuanensis seedlings to drought and rehydration, underscoring the central role of endogenous ethylene in drought tolerance. Ethephon treatment effectively enhanced ethylene accumulation and biosynthetic enzyme activity, thereby improving drought adaptability. These findings lay a theoretical foundation for subsequent molecular functional studies and the conservation biology of this endangered species. Full article

Background/Objectives: Stroke survivors with dysphagia are usually fed with different feeding routes ranging from oral to percutaneous endoscopic gastrostomy (PEG). However, the impact of the feeding route on the gastric myoelectric activity (GMA) is little-studied. This work examined the effect of feeding route on GMA changes in stroke survivors with dysphagia. Methods: This study included 50 patients (20% women) who were divided into three groups based on their feeding route: an oral group (n = 20), a nasogastric group (NGT) (n = 20), and a PEG group (n = 10). For all participants, a nutritional assessment was conducted, and the GMA was measured using a transcutaneous multichannel electrogastrogram (EGG) with a water load satiety test before and after water loading. The EGG-related parameters used in the analysis included the average power distribution by frequency region and the average dominant frequency (ADF). Results: The study sample experienced ischemic stroke (66%) or hemorrhagic stroke (34%). At the baseline phase, the PEG group exhibited significantly longer periods of normogastria compared to the NGT and oral groups. Moreover, protein intake was significantly higher in the PEG tube feeding group compared to the other groups. Based on the type of stroke, the ischemic stroke group showed significantly higher tachygastria periods during postprandial EGG recording (p = 0.022). The energy and protein consumptions were significantly higher in the hemorrhagic stroke group (p = 0.001, p = 0.028, respectively). Conclusions: The GMA pattern is distinctive for the type of stroke. The PEG feeding route showed more periods with normogastria and the best protein intake. Full article