Search Results (348)

Sarcopenia, characterized by the progressive loss of skeletal muscle mass and function, is exacerbated by glucocorticoid exposure. Although there is growing interest in natural therapies for muscle atrophy, the effects of Quercus acuta Thunb. fruit extract (QA) on sarcopenia or glucocorticoid-induced muscle loss had not been previously investigated. QA is an evergreen oak known for its antioxidant and anti-inflammatory properties, with polyphenolic components reported to enhance oxidative and metabolic homeostasis in various tissues. Based on these properties, we hypothesized that QA could counteract muscle atrophy by modulating anabolic and catabolic signaling pathways. The research utilized both in vitro (C2C12 myotubes) and in vivo (ICR mice) models to assess QA’s effects. Daily oral administration of QA (100–200 mg/kg) was given to mice with dexamethasone (Dex)-induced muscle atrophy. Techniques included H&E staining to assess muscle mass and fiber cross-sectional area (CSA), Western blot, and ELISA analyses to investigate signaling pathways. Confocal imaging was also used to confirm cellular changes. In vitro QA treatment improved myotube integrity by increasing myogenic differentiation markers (MyoD, MyoG) and suppressing atrophy-related E3 ligases, specifically MuRF-1 and FBX32/Atrogin-1. Confocal imaging showed that QA inhibited the nuclear localization of FOXO1 and reduced FBX32 expression. In vivo, daily oral administration of QA significantly preserved gastrocnemius muscle mass and fiber cross-sectional area in Dex-treated mice. QA restored the IGF-1/PI3K/Akt signaling pathway and attenuated FOXO1-dependent proteolytic activation. Collectively, these findings demonstrate that QA possesses potent anti-atrophic and myoprotective effects mediated through the modulation of the IGF-1/Akt-FOXO axis. QA has potential as a novel natural therapeutic for preventing glucocorticoid-induced sarcopenia. Full article

Osteoarthritis (OA) is a debilitating joint disease affecting over 500 million people globally, characterized by cartilage degradation, chronic pain, and failed tissue repair. Neurogenic inflammation, driven by neuropeptides including Substance P (SP) and calcitonin gene-related peptide (CGRP), plays a key role in the pathogenesis of OA. This study explores the therapeutic potential of extracellular vesicles (EVs) derived from infrapatellar fat pad mesenchymal stem/stromal cells (IFP-MSCs) transduced with CGRP antagonist CGRP_8-37 (aCGRP IFP-MSC EVs). These EVs are enriched in anti-inflammatory miRNAs and proteins, and they express neprilysin (CD10), enabling SP degradation. Herein, several LncRNAs were identified, which have been known to interact with miRNAs that affect the knee joint homeostasis. Specifically, 11 LncRNAs (ZFAS1, EMX2OS, HOTAIRM1, RPS6KA2-AS1, DANCR, LINC-ROR, GACAT1, GNAS-AS1, HAR1A, OIP5-AS1, TERC) interact with miRNAs that promote cell proliferation, prevent apoptosis, and preserve homeostasis. In vitro, aCGRP IFP-MSC EVs downregulated pro-inflammatory markers (TNF, TLR4, MAPK8) in dorsal root ganglia and promoted chondrocyte gene expression consistent with anabolism and matrix remodeling. In vivo, intra-articular EV delivery attenuated pain behaviors, preserved the cartilage structure, restored PRG4+ stem/progenitor cell localization, and trended toward reduced SP levels. Histological analysis confirmed improved collagen organization and reduced matrix degradation. These findings suggest that aCGRP IFP-MSC EVs exert multimodal effects on neuroinflammation, cartilage regeneration, and joint homeostasis. This cell-free, gene-enhanced EV therapy offers a promising disease-modifying strategy for the treatment of OA, with the potential to address both structural changes and chronic pain associated with this disease. Full article

No pharmacological interventions exist that can restore or preserve auditory function in the mammalian cochlea. Auditory hair cells (HCs) do not spontaneously regenerate, leading to permanent hearing loss. In non-mammalian vertebrates, HC regeneration happens through proliferation and differentiation of their clonally related supporting cells (SCs). The present study supports the potential of quinoxaline (Qx), a nonsteroidal anti-inflammatory compound, to stimulate SC proliferation in the auditory sensory epithelium, a process that may prime the tissue for future HC regeneration. We synthesized a series of Qx derivatives by introducing various substitutions, ranging from hydrophilic to lipophilic. Seventy analogs were generated and tested in vitro and in vivo. Among those, only one (Qx-100) exhibited the best medicinal chemistry profile and was further modified to expand the structure–activity relationship of the chemotype, develop additional analogs, and optimize potency, bioavailability, and in vivo efficacy. Ten new lead variants were generated. Of those, Qx-294 and Qx-301 demonstrated promising in vitro Absorption, Distribution, Metabolism, and Excretion (ADME) profiles and were selected for further testing. Overall, both compounds were rapidly absorbed in zebrafish and mice and promoted cell proliferation in vitro and in vivo without signs of apoptosis, supporting their potential for sensory HC regeneration. Full article

The use of phytobiotics in aquafeeds is a promising strategy to enhance performance and resilience to disease. This study evaluated the protective role of Morus alba (MA) extract against Vibrio cholerae, integrating in vivo responses in Dormitator latifrons (growth, biochemical and enzymatic responses, haemato-immunology and tissue histopathology) with in vitro assessment of V. cholerae growth, virulence-associated gene expression and cellular morphology. D. latifrons juveniles were fed five diets (0, 5, 10, 15 and 20 g/kg feed; three tanks per treatment, 15 fish per tank) for eight weeks, followed by a 7-day challenge with V. cholerae. MA increased growth and feed utilisation (p < 0.05); the 20 g/kg group reached 27.57 g final weight with a feed conversion ratio of 1.24, and whole-body protein and lipid contents rose at higher doses. MA modulated plasma biochemistry and key digestive (amylase, lipase), metabolic (ALT, AST) and antioxidant (SOD, CAT, GPx) enzymes, and improved haematological profiles. Histology of the intestine, liver and spleen showed preserved architecture and reinforced mucosal features in supplemented fish, particularly at 15–20 g/kg. Post-challenge, supplemented groups exhibited higher survival/relative protection than controls, alongside lower transaminases and stronger antioxidant responses. In vitro, MA extract inhibited V. cholerae growth, attenuated virulence-associated gene (toxR, ompU) expression and induced marked morphological damage in planktonic cells. Multivariate analyses (Z-score heatmaps and PCA) linked immune–enzymatic improvements with growth and protection. Overall, 15–20 g/kg MA optimised immunophysiological status and disease resistance, supporting MA as a functional feed additive for sustainable aquaculture of D. latifrons. Full article

Genome modification of legumes, peas in particular, is accompanied by significant challenges. Establishing a reliable reporter system to identify tissue that expresses foreign DNA may help to optimize and develop transformation protocols for these species. The RUBY system, based on the synthesis of red betalain from tyrosine, offers a convenient solution for monitoring the efficiency of transgene introduction. To evaluate the effectiveness of RUBY application in pea tissue culture, we combined agrobacterial transformation with an in vitro cultivation system, inducing callus development. Transformed explants demonstrated RUBY pigmentation, but it disappeared during cultivation. We hypothesized that this issue is caused by tyrosine depletion. To check this suggestion, we tested whether tyrosine supplementation could maintain RUBY coloring. In the later stages, pigmentation still could not be preserved. However, our modified conditions increased the percent of colored shoot apex explants during the early cultivation stages. Thus, it is likely that some explants transformed with the RUBY cassette do not synthesize a sufficient amount of betalain due to the deficit of endogenous tyrosine. In this case, adding exogenous tyrosine would enhance betalain production and improve the detectability of tissues containing the RUBY cassette. These data can be used for the optimization of RUBY application conditions for peas and other species. Full article

Cell migration plays a central role in physiological processes such as wound healing, tissue regeneration, and immune responses, as well as in pathological conditions like chronic inflammation and tumor metastasis. Among the in vitro approaches to study this phenomenon, the conventional wound healing assay (scratch assay) has been widely used due to its simplicity and low cost. However, its limitations, including poor reproducibility, damage to the extracellular matrix (ECM), and lack of dynamic physiological conditions, have prompted the development of microfluidic alternatives. Scratch-on-a-chip platforms integrate engineering and microtechnology to provide standardized, non-destructive methods for wound generation, preserve ECM integrity, and allow precise control of the cellular microenvironment. These systems also enable miniaturization, reducing reagent and cell consumption, while facilitating the application of biochemical or physical stimuli and real-time monitoring. This review synthesizes advances reported in the literature, addressing the different wound induction strategies (enzymatic depletion, physical depletion, and physical exclusion), the role of ECM composition, and the impact of mechanical forces such as shear stress. Overall, scratch-on-a-chip assays emerge as promising tools that enhance reproducibility, better mimic in vivo conditions, and broaden applications for therapeutic testing and mechanistic studies in cell migration. Full article

Lead (Pb) contamination severely threatens plant health and biodiversity, particularly in mining-affected ecosystems. The phytohormone, salicylic acid (SA), plays a crucial role in regulating plant stress responses. Here, the effect of SA supplementation on the in vitro response of Cistus heterophyllus subsp. carthaginensis, a critically endangered Mediterranean shrub, to Pb stress (50 μM Pb(NO₃)₂) was evaluated. SA dose pretreatment (100 μM) was selected based on phenolic accumulation in leaf tissues. Physiological and biochemical parameters—including mineral content, photosynthetic performance, total phenolics, and antioxidant activity—were quantitatively analyzed. SA pretreatment markedly reduced Pb accumulation (25%) while promoting Fe (73%), K (29%), and Mn (15%) uptake. It also alleviated Pb-induced photosynthetic impairment, preserved chloroplast integrity, increased chlorophyll content, and reduced the accumulation of lipid peroxidation products. Furthermore, SA promoted the accumulation of phenolic compounds—such as flavonoids, (+)-catechin, gallic acid, and hydroxycinnamic acid derivatives—in Pb-treated shoots, resulting in increased antioxidant capacity, as reflected by DPPH and FRAP assays, and protection against lipid autooxidation. However, no differential effect of SA pretreatment on DNA protection against oxidative damage was observed. Overall, SA acted as an effective priming agent, maintaining mineral homeostasis, photosynthetic stability, and antioxidant defense under Pb stress. These findings highlight its potential for enhancing plant resilience to Pb toxicity and for supporting the conservation and reintroduction of C. heterophyllus in contaminated habitats. Full article

Implantable scaffolds are increasingly recognized as transformative tools in regenerative medicine, offering the potential to prevent or mitigate tissue degeneration. Osteoarthritis is a widespread degenerative joint disease that often progresses from early focal lesions to severe joint damage, creating substantial clinical and socioeconomic burdens. Preventive strategies for early-stage lesions remain limited. This study reports the design and development of a functional polymeric scaffold intended to support early tissue regeneration and potentially prevent lesion progression. The scaffold consists of an electrospun poly (ε-caprolactone) nanofibrous membrane enriched with glycosaminoglycans, including hyaluronic acid and chondroitin sulfate, to mimic essential features of the cartilage extracellular matrix and provide a supportive microenvironment. Complete structural, physicochemical, and mechanical characterization was performed to assess the scaffold architecture, stability, hydration properties, and suitability for tissue environments. In vitro investigations were conducted to evaluate cytocompatibility and the interaction of the scaffold with relevant cell types. The scaffold is designed as a potential future preventive strategy to support cartilage integrity and limit disease progression. This approach represents a promising strategy to preserve joint integrity and function, addressing a critical unmet clinical need and enabling translation toward clinical application. Full article

Despite the substantial global impact of ischemic stroke, current therapeutic options remain limited and only partially effective. To advance neuroprotective strategies that could improve the safety and efficacy of existing treatments while preserving brain tissue, we synthesized and evaluated seven new nitrones (MC3, MC5, MC7) and oximes (MC1, MC2, MC4, MC6) derived from different neuroactive steroids—ethisterone (MC1–3), mifepristone (MC4–5) and stanolone (MC6–7)—in an in vitro model of cerebral ischemia. Overall, these derivatives exhibited neuroprotective and antioxidant effects superior to those of the reference compounds cholesteronitrone ChN2, α-tert-butyl nitrone (PBN) and N-acetylcysteine (NAC). Notably, nitrones showed greater neuroprotective, anti-necrotic, and antioxidant potency than their corresponding oximes, regardless of the degree of molecular conjugation. Among them, the stanolone-derived nitrone MC7, which lacks conjugated double bonds, displayed the most balanced and robust profile, consistently enhancing cell viability, reducing necrotic cell death, and suppressing superoxide anion production. Consequently, MC7 has been selected as a promising lead compound for further in vivo studies of cerebral ischemia. Full article

The high incidence of cardiovascular disease and the early failure of bioprosthetic valves due to calcification have driven the development of anti-calcification technologies. As a new storage technology, drying treatment is expected to delay the calcification process by reducing glutaraldehyde residues. However, the effects of drying treatment on the mechanical properties and valve functions of bovine pericardial materials are still unclear. The objective of this study is to evaluate the influence of drying and rehydration treatments on the mechanical integrity and geometric properties of bovine pericardium and the hemodynamic performance of bioprosthetic valves made with these tissues. Cross-linked bovine pericardial samples (n = 15) were divided into three groups—wet (control group progressed with normal glutaraldehyde), dehydrated (ethanol–glycerol dehydration), and rehydration (saline immersion) groups—and the geometric stability and nonlinear mechanical behaviors of the materials were analyzed via thickness measurements and uniaxial and biaxial tensile tests. Quantitative results showed that thickness remained stable across groups (wet: 0.356 ± 0.052 mm; dry: 0.361 ± 0.053 mm; rehydrated: 0.361 ± 0.053 mm, p > 0.05). Elastic modulus values were preserved (wet: 12.5 ± 1.8 MPa; dry: 13.1 ± 2.0 MPa; rehydrated: 12.7 ± 1.9 MPa, p > 0.05), and anisotropy ratio showed no significant changes (1.53 ± 0.06 vs. 1.57 ± 0.07, p > 0.05). The hemodynamic performance of bioprosthetic valves made with these materials was evaluated in vitro using a pulsating flow simulation. Hemodynamic parameters demonstrated excellent preservation: effective orifice area (wet: 2.625 ± 0.11 cm²; rehydrated: 2.585 ± 0.12 cm², Δ = 1.5%, p = 0.32) and regurgitation fraction (wet: 39.35 ± 2.9%; rehydrated: 42.78 ± 3.2%, p = 0.15) showed no statistically significant differences. The geometric properties of the material were not significantly changed by the drying treatment, and the material maintained its nonlinear viscoelastic characteristics and anisotropy. The rehydrated bioprosthetic valves did not differ significantly from those in the wet group in terms of the effective orifice area, regurgitation fraction, and transvalvular pressure difference, and the hemodynamic performance remained stable. Full article

Oxidative stress compromises follicle survival during in vitro culture. Natural antioxidants may reduce cellular damage and preserve tissue integrity. This study evaluated the ethanolic extract from Punica granatum L. (EE-PG) on bovine ovarian tissue cultured in vitro. Bovine ovarian (n = 24) fragments were cultured for 6 days in αMEM⁺ medium with or without EE-PG at 10, 50, or 100 µg/mL. At the end of the in vitro culture, the medium was used to evaluate antioxidant capacity (DPPH and ABTS assays), while the fragments were collected for morphological and biochemical analyses. HPLC-UV-Vis confirmed the presence of α-punicalagin in the pure EE-PG. At 100 µg/mL, EE-PG showed the strongest effects: it had higher antioxidant capacity, preserved follicle morphology and ultrastructure, and promoted follicle activation. At 50 and 100 µg/mL, the extract also reduced malondialdehyde (MDA) and increased thiol levels, indicating protection against lipid peroxidation. In contrast, 10 µg/mL had little effect. Follicle and oocyte diameters were not significantly altered, but the collagen I/III ratio increased at higher concentrations, suggesting extracellular matrix remodeling. Together, these findings demonstrate that EE-PG protects bovine preantral follicles from oxidative stress, maintains redox balance, and preserves tissue integrity. These results reinforce the potential of Punica granatum L. extract as a natural antioxidant in reproductive biotechnologies and fertility preservation. Full article

(1) Background: Most robotic MIS platforms lack native haptic feedback, leaving surgeons to infer tissue loads from vision alone—an especially risky limitation in esophageal procedures. (2) Methods: We develop a sensorless, image-only force-estimation pipeline that maps endoscopic video to tool–tissue forces using a lightweight EfficientNetV2B0 CNN. The model is trained on 9691 labeled frames from in vitro esophageal experiments and validated against an FT300 load cell. For intraoperative feasibility, the system is deployed as a plug-in on PARA-SILSROB, consuming the existing laparoscope feed and driving a commercial haptic device. The runtime processes every 10th frame of a 60 FPS stream (≈6 Hz updates) with ~15–20 ms per-prediction latency. (3) Results: On held-out tests, the model achieves MAE = 0.017 N and MSE = 0.0004 N², outperforming a recurrent CNN baseline while maintaining real-time performance on commodity hardware. Integrated evaluations confirm stable operation at the deployed update rate and low latency compatible with closed-loop haptics. (4) Conclusions: By avoiding distal force sensors and preserving sterile workflow, the approach is readily translatable and retrofit-friendly for current robotic platforms. The results support the practical feasibility of real-time, sensorless force feedback for robotic esophagectomy and related MIS tasks, with potential to reduce tissue trauma and enhance operative safety. Full article

Although nanotechnology is increasingly applied in plant tissue culture in many parts of Europe, its use in the Balkans remains limited, revealing a regional gap with untapped potential for advancing in vitro propagation and preservation of woody plant species. Building upon a recently published regional review covering 2001–2024, which analyzed in vitro biotechnology progress in nine Balkan countries, this paper introduces the concept of a “nano gap”, referring to the limited connection between existing nanotechnology research potential and its use in in vitro woody plant biotechnology. In Serbia, Greece, Bulgaria, Croatia, and Albania, significant progress has been made in optimizing micropropagation and in vitro conservation strategies by introducing temporary immersion systems, synthetic seed technology, adapting genotype-specific sterilization and multiplication protocols, and modifying established cryopreservation methods for regional woody species. However, the integration of nanotechnology into these systems remains largely unexplored. To date, there are no published results or validated applications for nano-enhanced media or nanoscale delivery systems for micropropagation and in vitro conservation of woody species. The limited integration of nanotechnology may be due to insufficient funding, lack of specialized infrastructure, and limited interdisciplinary expertise. Nevertheless, many Balkan countries possess growing capacities in nano-applications within agriculture and environmental sciences and are ready to advance toward interdisciplinary research and innovation. By mapping both scientific readiness and structural barriers, this review provides a strategic framework for bridging the “nano gap” and offers a novel regional perspective with broader implications for European research policy, sustainable agriculture, biodiversity preservation, and green innovation. Full article

Dry Eye Disease (DED) is a multifactorial condition of the ocular surface, with one potential cause being damage from eye drops containing preservatives such as benzalkonium chloride (BAC). Current treatments for DED are unsatisfactory; therefore, it is worth exploring new therapies based on the secretome derived from stem cells. Human stem cells are important sources of growth factors and cytokines that promote tissue regeneration. The secretome of these cells can be obtained in vitro in conditioned medium (CM). The aim of the study was to evaluate the effect of CM derived from adipose-derived stem cells (hADSCs) and amniotic membrane-derived cells expressing mesenchymal and/or epithelial markers on limbal stem cells (LSCs) damaged by BAC, focusing on their regenerative potential. The study used two experimental models: the first focused on neutralizing the toxic effects of BAC when each CM was administered concurrently, and the second on the therapeutic effects of CM after prior cell damage by BAC. The effects of CM on LSCs were assessed, including apoptosis, cell cycle progression, proliferation, migration, and inflammation. CM from ADSCs and amniotic cells were shown to significantly reduce BAC-induced damage to LSCs. All tested CM promoted LSC regeneration, although their efficacy varied among treatments. The application of CM during BAC exposure yielded stronger and more consistent benefits than post-injury treatment. Full article

Radiation-induced lung inflammation (RILI) is a major complication of thoracic radiotherapy, characterized by excessive inflammation and subsequent fibrosis that compromise pulmonary function and treatment outcomes. This study explores the pharmacological properties of a newly synthesized Lipoxin A4 analogue (CYNC-2) to mitigate RILI by modulating the AMP-activated protein kinase (AMPK)/NOD-like receptor family pyrin domain containing 3(NLRP3) inflammasome pathway. A murine RILI model was established in mice by delivering a single high-dose (ablative) X-ray irradiation to the left lung. Mice in the treatment group received CYNC-2 via tail-vein injection three times per week for 2 weeks. The effects of CYNC-2 on RILI were evaluated histological, immunohistochemical analysis of lung tissues, cytokine profiling, lung function testing using a FlexiVent system, and micro-computed tomography (micro-CT) imaging of lung damage. In parallel, two human lung cell lines—L132 (normal bronchial epithelial cells) and A549 (lung carcinoma cells)—were irradiated with 6 Gy X-rays and treated with CYNC-2 to assess cell viability and changes in AMPK/NLRP3 pathway markers via qPCR and immunofluorescence. Lung tissue sample from patients who underwent thoracic radiotherapy were also examined to validate key findings. CYNC-2 activated AMPK and inhibited mTOR signaling, which suppressed NLRP3 inflammasome activation and led to reduced secretion of pro-inflammatory cytokines (IL-1β, IL-6, and TGF-β1). In vitro, CYNC-2 mitigated radiation-induced inflammatory responses and preserved cellular viability. Overall, CYNC-2 effectively dampened acute pulmonary in the RILI model. These findings suggest that targeting the AMPK/NLRP3 inflammasome pathway via a stable LXA4 analogue such as CYNC-2 is a promising therapeutic strategy to improve clinical outcomes for patients receiving thoracic radiation therapy. Full article