Search Results (744)

Mesenchymal stromal cells (MSCs) are heterogeneous, multipotent cell populations that have become one of the most extensively investigated cell types in veterinary regenerative medicine. They can be isolated from a wide range of adult and perinatal tissues and have attracted considerable interest because of their immunomodulatory properties, broad secretory activity, and potential to promote tissue repair. However, increasing evidence indicates that MSCs should not be regarded as a uniform therapeutic product, as their biological behaviour is influenced by tissue source, donor characteristics, manufacturing conditions, and the recipient microenvironment. Likewise, their therapeutic effects are now understood to arise predominantly through dynamic interactions with the host, including inflammatory licensing, immunomodulation, secreted bioactive factors, extracellular vesicles, metabolic exchange, and responses to apoptotic cells, rather than through durable engraftment and direct tissue replacement alone. Although clinical evidence supports the use of selected MSC products in canine osteoarthritis and suggests promise for equine musculoskeletal disorders, most other veterinary applications remain preclinical or investigational. This review critically examines current knowledge of MSC biological identity, tissue-source diversity, molecular mechanisms, therapeutic applications, and comparative translational roles in companion and farm animals. It also discusses the challenges that continue to limit clinical translation, including biological heterogeneity, product characterization, potency assessment, manufacturing standardization, and regulatory considerations, and highlights future directions for the development of safe, effective, and evidence-based MSC therapies in veterinary medicine. Full article

Biodegradable alginate films containing silver nanoparticles (AgNPs) are being increasingly investigated as active agricultural materials (e.g., antimicrobial mulches) and active food packaging. However, since these materials ultimately degrade in soil after use, assessing their environmental compatibility and potential phytotoxicity upon release is crucial. The aim of this study was to synthesize films containing AgNPs in sodium alginate using xylose as a reducing agent and to determine their effect on culturable rhizosphere microorganisms and selected biochemical parameters in young arugula (Eruca vesicaria L. subsp. sativa) plants. Alginate films containing three nominal AgNP loadings (50, 100, and 150 mg L⁻¹) and a control film without AgNPs were synthesized. The films were cut into square pieces (4 cm²) and placed in 0.076 L multipots filled with peat substrate, into which arugula seeds were sown. During the experiment, the abundance of culturable rhizosphere bacteria and fungi was determined, and the young arugula plants were subjected to biochemical analyses. The results showed that the AgNP-containing films did not significantly affect the abundance of bacteria and fungi in the rhizosphere under the conditions tested. The tested films also did not markedly alter the measured parameters in the tissues of young arugula plants, including ascorbic acid, photosynthetic pigments, sugars, dietary protein, and glutathione. However, they reduced phenolic content, altered antioxidant activity, and led to detectable silver accumulation in plant tissues, especially at the highest nominal AgNP loading (150 mg L⁻¹). These findings indicate limited but selective biochemical effects during the early growth stage of arugula rather than a complete absence of plant response. Full article

Total flavonoids (TF) from Artemisia argyi, a traditional edible-medicinal Asteraceae herb, were extracted via a food-grade ternary deep eutectic solvent (TDES, Glycerol/Levulinic Acid/Xylitol = 1:1:1 molar ratio, 30% w/w water) under ultrasound assistance. A hybrid response surface methodology–genetic algorithm–artificial neural network (RSM-GA-ANN) model optimized parameters to deliver a maximum TF yield of 107.7 mg/g—1.5–1.8-fold higher than conventional hydroalcoholic extraction. Greenness was quantified by AGREEprep (score = 0.70) and MoGAPI (score = 80/100), confirming the method’s sustainability, with TDES retaining >84% extraction efficiency over three reuse cycles. Molecular dynamics simulations revealed TDES forms a more stable hydrogen bond network with plant cell walls (average H-bond lifetime: 101.279 ps vs. 46.698 ps for 50% ethanol), a finding validated by density functional theory calculations showing TDES establishes 7–9 hydrogen bonds with cellobiose (the cellulose repeating unit), far exceeding ethanol’s 1–2 hydrogen bonds. Purified TF exhibited potent radical-scavenging activity (DPPH IC₅₀: 0.176 mg/mL; ABTS IC₅₀: 0.159 mg/mL) and multipotent enzyme inhibition (α-glucosidase IC₅₀: 55.31 μg/mL; acetylcholinesterase IC₅₀: 0.618 mg/mL; pancreatic lipase IC₅₀: 0.125 mg/mL). TF also suppressed HCT116, A549, and HepG2 proliferation (IC₅₀ ≈ 50 μg/mL) and protected HepG2 cells against H₂O₂-induced oxidative damage. As an in silico probe, the predominant quantified flavonoid eupatilin (3.64 mg/g) docked to xanthine oxidase (−7.78 kcal/mol vs. allopurinol −6.86), offering a structural hypothesis for XO interaction without attributing mixture bioactivity to a single compound. This TDES-based platform offers a scalable, green route to valorize A. argyi for functional food and nutraceutical applications. Full article

The bone marrow stromal microenvironment is damaged in patients with acute leukemia. The aim of this study was to analyze changes associated with the extracellular matrix, mitochondrial function, and vesicular transport in the proteome of multipotent mesenchymal stromal cells (MSCs) in patients at the onset, in remission, before, and 1–3 months after allogeneic hematopoietic stem cell transplantation (allo-HSCT). The study included paired MSCs samples from the bone marrow of 12 patients at the onset and in remission of acute leukemia (4 ALL, 8 AML) and eight patients before and after allo-HSCT (4 ALL, 4 AML). MSCs from eight healthy donors were used as a control. The growth characteristics and the proteome subsets describing extracellular matrix, mitochondria, and vesicular formation were studied. The proteome of the patients’ MSCs differed significantly from that of the donor MSCs, both at the onset and in remission. Changes noted in the composition of extracellular matrix proteins may affect cell adhesion and access to growth factors. Significant changes were revealed in proteins affecting mitochondrial function. Vesicular transport proteins also differed between the donor and patient groups. Unexpectedly, no differences were found between the MSCs of donors and patients before and after allo-HSCT. Full article

Janus nanoparticles (JNPs), named after the bi-faced Roman God has emerged as a hot topic in the present era of nanoscience because of their chemical, structural and physical uniqueness. These particles stand out in the crowd as they are composed of two or more faces with contrasting functional properties in the same molecule. The need for two or more functions in a single molecule that can be used in chemical, biological or physical fields, such as delivering drugs combined with imaging, is the need of the hour and JNPs open gates for addressing this issue as these self-tailored particles have found their application in various in vivo and in vitro domains. However, despite their vivid application, a larger sector of application still needs to be explored. Synthesis methods involve masking, self-assembly and microfluidics, and comparative analysis of these methods, listing their pros and cons, would assist in overcoming the difficulties in the commercialisation of these particles. Moreover, the systematic analysis of differences in their structure with reference to their functionality and characterisation methods would lead us to a better understanding of the subject. This review discusses the various synthesis strategies and their comparison, the anisotropic nature of the JNPs conferring various distinguished properties, their application in emulsion stabilisation, bio-imaging, drug-targeting, drug-delivery, and biosensing domains, the characterisation methods involved, challenges and future aspects. The future aspects section maps a few hypotheses that might be useful in expanding the horizons of usage. The novelty of this review lies in the critical analyses of the synthesis methods and characterisation. Each type of JNP has been analysed for its advantages and limitations, and probable hypotheses to address the existing challenges have been jotted down. Full article

Early hematopoietic injury is a critical determinant of hematopoietic acute radiation syndrome (H-ARS), yet biologically relevant cellular endpoints and optimal evaluation windows for therapeutic screening remain poorly defined. Here, we combined high-dimensional mass cytometry (CyTOF) and flow cytometry to characterize early hematopoietic stem and progenitor cell (HSPC) remodeling following 6.5 Gy total-body irradiation. Radiation exposure induced rapid bone marrow injury, with substantial cellular loss and reduced viability occurring within hours after irradiation. CyTOF analysis revealed that radiation-induced injury was characterized by selective remodeling rather than uniform depletion of the HSPC compartment. While long-term hematopoietic stem cells (LT-HSCs) were relatively preserved, short-term HSCs (ST-HSCs), multipotent progenitors (MPPs), and megakaryocyte–erythroid progenitors (MEPs) exhibited marked reductions during the early phase after irradiation. Importantly, surviving cells retained partial differentiation capacity during this period, indicating that the early post-irradiation phase represents a biologically informative window for therapeutic evaluation. These radiosensitive populations were subsequently validated using a simplified flow cytometry platform and remained detectable under short-term in vitro culture conditions. Collectively, our findings identify key radiosensitive HSPC subsets and establish an early hematopoietic injury framework that integrates optimal evaluation timing with practical cellular endpoints for H-ARS therapeutic screening and radiomitigator development. Full article

Mesenchymal stromal cells (MSCs) are a versatile platform for regenerative medicine and gene delivery because they combine multipotency, immunoregulatory activity, and injury-directed trafficking. Translation is nevertheless limited by donor- and tissue-dependent heterogeneity, variable biodistribution, and engineering-related risks. This review evaluates genetically modified MSCs as medicinal products rather than as a general MSC class. We compare self-inactivating lentiviral (SIN-LV) transduction, which provides efficient and durable expression and has limited early clinical experience, with targeted genome editing, which can define the integration locus and copy number but remains constrained by variable precise knock-in efficiency, off-target and double-strand-break-associated effects, manufacturing cost, and the absence of long-term clinical safety data. We integrate preclinical and clinical evidence with GMP-compatible manufacturing, potency testing, genomic surveillance, and release criteria. Particular attention is given to safe-harbor integration and B2M/CIITA-based hypoimmunogenic designs as strategies to reduce engineering-related batch variability and HLA-dependent donor variability. Together, these developments support a transition from empirically optimized MSC preparations toward molecularly defined cellular medicines with predefined genotype, expression, potency, and safety attributes. Full article

Background/Objectives: Mesenchymal stem cells (MSCs), due to their regenerative and multipotent properties, have emerged as promising therapeutic agents in tissue repair and regeneration. These biological characteristics might contribute to optimized anastomotic healing and to a reduction in postoperative complications following digestive surgery. The present study aimed to evaluate whether intraperitoneal or perianastomotic administration of MSCs provides superior healing outcomes in colonic anastomoses in Wistar rats. Methods: MSCs were isolated from inguinal adipose tissue harvested from 2 Wistar rats. Thirty male Wistar rats were allocated to 3 groups: (i) the control group, with regular anastomosis, (ii) peri-anastomotic injection of MSCs, and (iii) intraperitoneal injection of MSCs. The animals were sacrificed on postoperative day 14. The evaluated outcomes included clinical evolution, adhesion index, histological characteristics, and tissue hydroxyproline content. Results: The incidence of anastomotic leakage and the mortality rate were 0%. Therefore, the present study primarily demonstrates changes in surrogate markers of healing, including inflammatory response, collagen deposition, adhesion formation, and hydroxyproline content. The adhesion index was similar in the groups receiving MSC administration (p = 0.05); however, intraperitoneal administration demonstrated superior outcomes when compared to standard anastomosis in reducing adhesion formation (p = 0.002). Histopathological analysis showed a decreased inflammatory process and an increased collagen deposition at the anastomotic site following MSC administration (p < 0.05). Moreover, tissue hydroxyproline levels were significantly increased after both perianastomotic (0.831 ± 0.02, p < 0.05) and intraperitoneal (0.54 ± 0.02, p < 0.05) MSC administration compared with the control group (0.251 ± 0.006). Conclusions: These results suggest that MSC administration may improve histological and biochemical markers associated with colonic anastomotic healing in a non-ischemic experimental model. The experimental model used is suitable for further studies aimed at determining the optimal indications, routes of administration, and adjunctive agents that may potentiate the effects of MSCs. Full article

Adult bone marrow-derived mesenchymal stem cells (BMSCs) are multipotent progenitors capable of differentiating into diverse cell lineages, including osteogenic, chondrogenic, adipogenic, and neuronal lineages. In BMSCs, intracellular glutathione (GSH) is a critical determinant of stemness maintenance and differentiation outcomes. However, how intracellular GSH homeostasis is regulated during BMSC-to-neuron differentiation remains unclear. In neurons, GSH synthesis critically depends on cysteine uptake mediated by the excitatory amino acid carrier 1 (EAAC1). Here, we investigated the expression, subcellular localization, and functional contribution of EAAC1 and its regulatory protein, glutamate transporter-associated protein 3-18 (GTRAP3-18) in mouse BMSCs and neuron-like BMSCs generated by Notch intracellular domain-based induction (NICD-3F BMSCs). BMSCs exhibited higher intracellular GSH levels than NICD-3F BMSCs, despite comparable levels of EAAC1 protein. In contrast, EAAC1-dependent cysteine uptake and plasma membrane localization of EAAC1 were markedly reduced in BMSCs, indicating differentiation-dependent regulation of EAAC1 trafficking. Treatment with the xCT inhibitor erastin reduced intracellular GSH levels in both BMSCs and NICD-3F BMSCs. GTRAP3-18 expression was high in BMSCs and significantly reduced in NICD-3F BMSCs. Notably, GTRAP3-18 knockout decreased intracellular GSH levels in BMSCs without altering total EAAC1 protein or intracellular cysteine levels, whereas in NICD-3F BMSCs, both GSH and EAAC1 protein levels were increased. These findings demonstrate lineage-dependent divergence in GSH regulatory mechanisms and reveal previously unrecognized functions of GTRAP3-18 in redox control during stem–to–neuron differentiation. Full article

Mesenchymal progenitor cells (MPCs) play a significant role in articular cartilage homeostasis and regeneration. Yet, the functional dynamics and molecular characteristics of MPCs may differ significantly across various pathological conditions. Hence, this study comprehensively investigates the biological and molecular characteristics of MPCs isolated from articular cartilage of patients with osteoarthritis (OA) and rheumatoid arthritis (RA), aiming to uncover disease-specific differences that could offer insights into targeted regenerative therapies. Using flow cytometry, gene expression analysis, and in vitro differentiation assays, we assessed the phenotype, growth potential, senescence, cytogenetic instability, and chondrogenic potential to delineate molecular pathways uniquely active in each disease context. Phenotypically, both OA and RA-MPCs retained markers of mesenchymal stem cells (MSCs), but OA-derived MPCs exhibited higher fold expression of progenitor markers (OCT-4, NANOG, SOX-2, and SSEA-4), suggesting a more activated state. Functionally, OA-MPCs demonstrated increased growth kinetics (higher proliferation rate and decreased population doubling time) with a significant shift towards adipogenic lineages (increased fold expression of LPL, AP2, and PPAR-γ). However, there were no differences in the osteogenic and chondrogenic potential. Gene expression analysis revealed upregulation of genes involved in extracellular matrix production and cartilage development (COL2-α1, ACAN, FGFR3, TGF-β3, ANXA6, CNTN1, MATN1, TGF-β1, VIM, and SOX9) in 3D cultures compared with 2D or monolayer cultures. Collectively, these findings demonstrate that, while multipotent MPCs are present in both OA and RA articular cartilage, they can exhibit fundamentally altered biological behaviors and molecular signatures reflective of the local disease microenvironment. Understanding these differences is critical for optimizing cell-based therapeutic strategies tailored to each condition and may facilitate the development of novel interventions targeting endogenous progenitor cells for cartilage repair. Full article

Hair follicle (HF) stem cells are multipotent adult stem cells that play a key role in the hair follicle cycle. However, it remains poorly understood how the outer root sheath (ORS)—specifically, the stem cells in the bulge region of the hair follicle—promotes skin repair. This study aims to investigate the role of bulge stem cells in tissue growth and repair and to determine whether the ORS of transplanted hair follicles can facilitate skin repair. We further seek to elucidate the mechanisms by which bulge stem cells contribute to hair follicle development, regeneration, and skin wound healing. In this study, hair follicle samples were obtained from neonatal mice using microdissection. Hair follicle morphology was assessed by Sirius red staining, H&E staining, and transmission electron microscopy. Immunofluorescence staining was used to detect changes in CD34 and SOX9 protein expression. Additionally, microdissection-based hair follicle transplantation and Western blotting were employed to investigate protein activation and inhibition in the Wnt/β-catenin signaling pathway. The results show that the hair follicle bulge, inner root sheath, and dermal papilla all increase in size as hair follicles grow, with each structure growing relatively rapidly on day 7. Treatment with Teplinovivint effectively inhibits the expression of Wnt/β-catenin signaling pathway-related proteins and hair follicle stem cell markers. Damaged hair follicle tissues cultured in vitro are capable of self-repair. At the transplantation site, the skin gradually closes as the outer root sheath wound heals. In contrast, the central region of the outer root sheath becomes progressively filled with numerous dividing cells and extracellular matrix. The inner portion of the outer root sheath is densely populated with cells, and the markers CD34 and SOX9 are also widely distributed. This indicates that activation of the Wnt/β-catenin signaling pathway enhances the proliferation and differentiation of hair follicle stem cells, thereby promoting hair follicle growth, repair of damaged follicles, and healing of skin wounds. Furthermore, this study demonstrates the feasibility of using transplanted outer root sheath (ORS) to repair skin wounds—specifically, the potential to achieve large-scale hair regeneration from a limited number of hair follicle stem cells—providing a new approach for the clinical treatment of skin injury disorders. Nevertheless, achieving long-term, stable, and scalable clinical translation of ORS stem cells for hair follicle regeneration remains a major challenge. Full article

Nervous system and corneal disorders are major causes of permanent disability worldwide, largely due to the limited regenerative capacity of ectoderm-derived tissues. Therefore, the development of accessible and ethically acceptable cell-based therapies promoting the repair and regeneration of these tissues is of considerable translational importance. In this study, we aimed to comparatively evaluate the ectodermal differentiation potential of human adipose-derived stem cells (ADSCs) and human amniotic epithelial cells (hAECs) in vitro, with hAECs serving as a reference cell population with established ectodermal plasticity. Primary ADSCs and hAECs were characterized phenotypically using flow cytometry and functional differentiation assays. Cells were subjected to a directed ectodermal differentiation protocol and assessed via morphological analysis, immunostaining for ectoderm-associated proteins, and RT-qPCR analysis of lineage-specific genes. ADSCs exhibited morphological changes following differentiation, including a more epithelial-like phenotype and an increased nucleus-to-cytoplasm ratio. Immunostaining revealed the induction of nestin and OTX2 expression after differentiation, which was particularly pronounced in ADSCs. Gene expression analysis demonstrated statistically significant upregulation of the ectoderm-related genes EN2, SOX1, and PAX6 exclusively in hAECs. Results suggest that in ADSCs the differentiation process was only partially activated. In conclusion, our findings further support the suitability of hAECs as a reference cell line for studies investigating ectodermal differentiation protocols, while also demonstrating that ADSCs exhibit a limited but detectable capacity for acquiring ectoderm-specific characteristics under defined in vitro culture conditions. Full article

Background: Human dental pulp stem cells (hDPSCs) are a promising source of multipotent mesenchymal stem cells (MSCs) for regenerative neurology because of their inherent neurogenic potential. However, robust and reproducible protocols for driving their terminal neuronal maturation in a fully defined, xeno-free environment are lacking. Methods: hDPSCs were isolated from a donor tooth and characterized for mesenchymal (CD105, CD90, CD73, CD13) and stemness-associated markers (SOX2, Oct3/4 and Nanog). Cells were differentiated in a novel, fully chemically defined medium 1% ITS medium (ITS: Insulin, Transferrin, Selenium) supplemented with glial cell line-derived neurotrophic factor (GDNF) or brain-derived neurotrophic factor (BDNF). Neuronal commitment and partial maturation were assessed via immunofluorescence, Western blot, and RT-PCR for markers such as NeuN (Neuronal nuclei) and NF-M (Neurofilament medium chain), and functionally by whole-cell patch-clamp electrophysiology. Results: Although undifferentiated hDPSCs expressed neural progenitor markers (βIII-tubulin and Nestin), only GDNF treatment in a chemically defined medium significantly upregulated mature neuronal markers (NeuN and NF-M) and downregulated mesenchymal markers. Importantly, GDNF-treated cells exhibited key functional changes, including hyperpolarized resting membrane potentials, increased membrane capacitance, and elevated input resistance, which are electrophysiological hallmarks of neural precursor or early neuronal maturation, compared to control cells cultured in medium containing fetal bovine serum (FBS). Although action potentials were not elicited, this represents a significant advancement toward achieving a functional neuronal state. Conclusion: This study demonstrates that a fully chemically defined medium enables GDNF to drive hDPSCs beyond the neural progenitor state towards a partially mature neuronal phenotype. This defined medium protocol eliminates serum variability, enhances reproducibility, and provides a critical step towards standardizing hDPSC-derived neuronal cells for disease modeling and cell-based therapy. Full article

Mesenchymal stromal cells (MSCs) are multipotent cells characterized by their regenerative capacity and strong immunomodulatory properties. In recent years, MSC-based therapy has attracted significant attention as a potential treatment for a wide range of immune-mediated and degenerative diseases. The therapeutic effects of MSCs are primarily mediated through paracrine signaling and secretion of cytokines that regulate immune responses and promote tissue repair. This review focuses on five key cytokines involved in MSC immunomodulation: interleukin-6 (IL-6), interleukin-10 (IL-10), transforming growth factor-beta (TGF-β), tumor necrosis factor-alpha (TNF-α), and interleukin-1 beta (IL-1β). These cytokines interact within a complex signaling network that allows MSCs to suppress excessive inflammation and restore immune balance. The role of MSC therapy is examined in several clinically relevant conditions, including systemic lupus erythematosus, systemic sclerosis, ischemic stroke, spinal cord injury, diabetes mellitus, and female infertility. Across these diseases, MSCs demonstrate the ability to inhibit pro-inflammatory immune cell activity, promote regulatory immune phenotypes, reduce oxidative stress, and stimulate regeneration through the secretion of growth factors and extracellular vesicles. Despite promising experimental and early clinical findings, several limitations remain, including variability in MSC sources, limited cell survival after transplantation, and the need for optimized dosing strategies. Overall, MSC therapy represents a multifunctional therapeutic approach combining immunomodulation, anti-inflammatory activity, and regenerative support. Further research is required to better understand cytokine interactions, improve standardization of MSC-based treatments, and enhance clinical efficacy across diverse pathological conditions. Full article

Mesenchymal stem/stromal cells (MSCs) are multipotent progenitor cells with potent immunomodulatory properties, making them attractive candidates for treating inflammatory and autoimmune diseases. A key mediator of MSC-induced immunosuppression is programmed death-ligand 1 (PD-L1), a checkpoint molecule that interacts with PD-1 on immune cells to regulate immune responses and promote tolerance. This review synthesizes current evidence on the role of PD-L1 expression in MSCs, emphasizing its effects on both the innate and adaptive immune systems, its therapeutic potential, and its utility as a biomarker for MSC potency and clinical efficacy. We examine how PD-L1 modulates T cell activation, dendritic cell maturation, macrophage polarization, and cytokine profiles, including its role in exosomal contexts. Additionally, we highlight its synergistic interactions with other immune checkpoints and discuss its dual function as both a therapeutic effector and a dynamic biomarker. Finally, we explore its relevance in clinical contexts such as autoimmune diseases, graft-versus-host disease, sepsis, and transplantation and conclude with a discussion of challenges and future directions in harnessing PD-L1 for MSC-based therapies. Full article