Search Results (1,336)

Adipose-derived stem cells (ADSCs) are widely used in regenerative medicine and are considered key effectors underlying the therapeutic efficacy of autologous fat grafting for scarring and skin fibrosis, yet clinical outcomes remain variable. This review examines how obesity alters the adipose microenvironment through chronic inflammation and metabolic dysfunction, resulting in epigenetic changes, mitochondrial impairment, oxidative stress, and premature cellular senescence in ADSCs. ADSCs from obese individuals exhibit reduced stemness, impaired differentiation, and a pro-inflammatory secretome with diminished regenerative capacity. While weight loss may partially reverse these effects, persistent epigenetic and functional memory limits full recovery. This review argues that donor metabolic status is a determinant of ADSC therapeutic potency and discusses key challenges and opportunities for improving regenerative outcomes. Full article

Background/Objectives: An ideal bone scaffold should promote bone cell growth and functional vascularization, hence the importance of imbuing biomaterials with pro-angiogenic cues. In this work, silica-based bioactive glasses, either pristine (SBA3) or doped with copper (SBA3_Cu), were embedded in poly(ε-caprolactone) (PCL), which was also used as a control. Methods: In vitro co-cultures of adipose-derived mesenchymal stem/stromal cells (ASCs) and human microvascular endothelial cells (HMEC-1s) were kept in α-MEM, MCDB131, and EndoGRO media to test the biomaterials. The co-cultures were visualized by immunofluorescence and SEM, while flow cytometry was performed to characterize cellular immunophenotype. The angiogenic potential was evaluated using conditioned media of co-cultures to perform a tubulogenesis assay and VEGF-A quantification. Results: Immunophenotypic analysis showed a significant decrease in the endothelial CD31+ cellular subset, whereas the OB-like cellular subset expressing CD105, CD73, CD90, and ALP increased in all culture media over time. In α-MEM, HMEC-1s were unable to form a capillary network independent of the substrates. A more organized network was visible when co-cultures were plated on PCL, in MCDB131 and EndoGRO, or if they were kept in EndoGRO on PCL/SBA3_Cu. The VEGF-A concentrations were similar in the conditioned media from co-cultures grown on PCL/SBA_Cu, in EndoGRO, and on PCL and PCL/SBA3, in MCDB131. Conclusions: The presence of copper did not promote the angiogenic potential of HMEC-1, likely due to the low concentration of released copper ions and the predominant osteoinductive effect of the other ions released by the bioglass. A re-evaluation of formulation and structure of bioglass scaffold could enhance the angiogenic potential. Full article

Background: Aging, especially after menopause, reduces the quantity and function of adult stem cells. Estrogen deficiency impairs proliferation, differentiation, and regenerative capacity. This study evaluated whether estrogen enhances endothelial differentiation of adipose-derived stromal cells (ADSCs) and improves therapeutic efficacy in critical limb ischemia (CLI). Methods: Male-derived ADSCs were assessed in vitro for endothelial differentiation using flow cytometry, biochemical assays, and angiogenesis analyses. Therapeutic effects were tested in a rat CLI model using endothelial-differentiated ADSCs (ED-ADSCs) with or without 17β-estradiol (E2). An ovariectomized (OVX) model examined estrogen deficiency and supplementation in vivo. Results: E2 promoted endothelial differentiation, increasing ERα/ERβ expression and activating PI3K/Akt/eNOS and MAPK signaling. This led to elevated VEGF expression, enhanced tube formation, and increased CD34⁺, KDR⁺, and CD31⁺ cell populations. In vivo, E2-pretreated ED-ADSCs significantly improved blood flow recovery. Estrogen deficiency reduced endothelial progenitor populations, which were restored by E2 supplementation. Conclusions: Estrogen modulates endothelial-associated characteristics and angiogenesis-related responses of ADSCs via ER-associated signaling, and may contribute to improved functional outcomes in ischemic conditions. E2 preconditioning may represent a potential strategy for stem cell-based therapy in estrogen-deficient settings. Full article

Background: Apoptotic vesicles (ApoVs) derived from adipose stem cells (ASCs) have recently emerged as important mediators of tissue repair and are implicated in pathways relevant to hypertrophic scar (HS). Although ASCs exhibit potential in scar modulation, the therapeutic value of their apoptotic clearance products remains largely unexplored. Methods: In this study, we investigated the efficacy and mechanism of staurosporine (STS)-induced adipose stem cell derived apoptotic vesicles (ASCs-ApoVs) in mitigating HS. Western blot, RT-qPCR, and immunofluorescence were used to assess fibrotic markers including α-SMA, COL1A1, and COL3A1 and so on in hypertrophic scar derived fibroblasts (HS-fibroblasts). Results: ASCs-ApoVs significantly reduced profibrotic marker expression in HS-fibroblasts without short-term cytotoxicity. CDC20 down-regulation was identified as a critical target, through which ASCs-ApoVs suppressed Wnt/β-catenin signaling, as evidenced by the downregulation of β-catenin, c-MYC, Cyclin D1, and AXIN2. The efficacy of ASCs-ApoVs in hypertrophic scar regulation was also confirmed by the rabbit ear scar model. Furthermore, ASCs-ApoVs demonstrated notable structural and functional stability. Conclusions: In summary, our results established STS-induced ASCs-ApoVs as a potent multi-target strategy for hypertrophic scar regulation. Besides, the scalable production, functional stability, and favorable safety profile of ASCs-ApoVs underscore a strong promise for clinical translation. Full article

Preterm labor is a major cause of neonatal morbidity and mortality and is frequently driven by infection-associated inflammation that promotes premature uterine activation. In this study, we investigated the effects of adipose stem cell-derived extracellular vesicles (ASC-EVs) on macrophage-mediated inflammatory signaling in uterine smooth muscle cells (HUtSMCs). An in vitro model was established by treating HUtSMCs with conditioned media derived from LPS-stimulated RAW264.7 macrophages. Activation of signaling pathways was assessed by Western blotting and immunofluorescence, and functional responses were evaluated using calcium flux and collagen gel contraction assays. Conditioned media from LPS-stimulated macrophages induced robust activation of MAPK (ERK1/2 and JNK) and NF-κB signaling, accompanied by IκB degradation and nuclear translocation of phosphorylated p65, whereas ASC-EVs pretreatment significantly attenuated these responses and reduced the expression of pro-inflammatory cytokines, including IL-6, IL-8, and MCP-1. Furthermore, macrophage-conditioned media enhanced intracellular calcium flux and contractile activity in HUtSMCs, both of which were suppressed by ASC-EVs. Inhibition of TLR4 signaling in macrophages reduced the inflammatory potency of conditioned media, indicating a key upstream role of macrophage TLR4 activation. Collectively, these findings demonstrate that ASC-EVs suppress macrophage-mediated inflammatory activation and downstream contractile responses, suggesting their potential as a cell-free therapeutic strategy for preventing inflammation-associated preterm labor. Full article

Objective: Effectively eliminating xenoimmunogenicity and achieving recellularization in cardiac xenografts remains a critical challenge in developing an ideal implantable xenograft. We have previously demonstrated that the removal of major antigens, including Galα1-3Gal (α-Gal) epitope and non-human sialic acid N-glycolylneuraminic acid (Neu5Gc), using α-galactosidase and peptide N-glycosidase F (PNGase-F), enables a synergistic effect with decellularization, significantly reducing the expression of carbohydrate-binding lectins without altering the biomechanical properties of the graft. The aim of this study was to establish an effective method for in vitro recellularization by seeding human mesenchymal stem cells (MSCs) on decellularized cardiac xenografts that had undergone optimal xenoantigen removal using α-galactosidase and PNGase-F. Additionally, this study aimed to evaluate the potential for in vivo recellularization. Methods: Decellularized porcine pericardium scaffolds treated with both enzymes were further modified by forming a fibrin mesh on their surface and within their structure, followed by the attachment of heparin and human vascular endothelial growth factor to the mesh. Subsequently, the scaffolds were seeded with human adipose tissue-derived stem cells for 8 weeks. In vitro recellularization, differentiation, and extracellular matrix remodeling of decellularized and enzyme-treated xenografts were assessed using vimentin, calponin, fibronectin, CD31, VWF, and phalloidin staining. To evaluate the potential for in vivo recellularization, decellularized glutaraldehyde-crosslinked xenografts with anticalcification treatments were seeded with rat bone marrow MSCs and implanted into rats subcutaneously to evaluate cell infiltration and calcification via histology, von Kossa staining, and micro-computed tomography. Results: In decellularized xenografts treated with both enzymes, stronger signals were detected and mesenchymal cell infiltration into the tissue was significantly faster, leading to accelerated recellularization. This recellularization process was more pronounced as time went on, with greater cell infiltration and evidence of cell differentiation. An in vivo study showed that decellularization and anticalcification treatments revealed stronger vimentin staining in histological analysis. The recellularization for our biocompatible scaffolds exhibited a lower degree of calcification compared to the non-recellularized tissue. Conclusions: We successfully developed major xenoantigen-free scaffolds by demonstrating the safety and synergistic effect of α-galactosidase and PNGase-F treatments and proved, for the first time, the effectiveness of recellularization using a human MSC monoculture on xenoantigen-free scaffolds. Furthermore, there was potential for in vivo recellularization of our biocompatible scaffolds seeded with MSCs. Full article

Bladder outlet obstruction (BOO) may lead to detrusor decompensation through progressive bladder remodeling. Most experimental studies rely on acutely induced BOO in rodents. Since progressive obstruction better reflects the condition, non-lethal models are needed to investigate chronic obstruction pathophysiology and evaluate regenerative therapies. This exploratory study aimed to evaluate (1) a progressive BOO model induced by an artificial urethral sphincter (AUS) in 2 dogs and (2) the systemic administration of autologous adipose-derived mesenchymal stem cells (ADMSCs) after obstruction release. Two intact male dogs underwent progressive BOO through gradual AUS inflation. Longitudinal assessment included telemetric urodynamic monitoring, urethral pressure profilometry, ultrasonography, post-void residual measurement, oxidative stress markers in serial blood samples and serial bladder biopsies for histology, transmission electron microscopy, immunohistochemistry, RT-qPCR and RNA sequencing (CCL2, CCR2, GFAP, VEGF, HGF). After AUS removal, one dog received three intravenous injections of 20 × 10⁶ PKH26-labelled autologous ADMSCs. BOO induced functional changes (increased detrusor pressure and urethral resistance, decreased urinary flow, prolonged voiding). No detrusor decompensation or fibrosis comparable to the human condition developed, encouraging refinement of this model. ADMSCs appeared to reach the bladder wall safely, but any influence on the glutathione redox system and CCL2 protein expression needs to be confirmed. Full article

Osteoporosis is an age-related metabolic bone disorder characterized by an imbalance between bone resorption and formation. Natural polyphenols have gained attention as potential complementary strategies for its prevention. In this study, we investigated the effects of a sustainable, polyphenol-rich extract from red grape pomace (GPE) on human mesenchymal stem cell (MSC) fate and its underlying mechanisms of action. We found that GPE significantly promoted osteogenic differentiation while suppressing adipogenic differentiation in canonical bone marrow-derived MSCs (BMSCs). This biological effect was preserved in adipose tissue-derived MSCs (AdMSCs) obtained from elderly patients (>65 years) at high cardiovascular risk. Mechanistically, GPE downregulated adipogenic markers (PPARγ, CD36 and FABP4) and enhanced osteogenic markers (RUNX2, ALP, OSX, BMP-2, OPN, COL1A1 and OCN). Moreover, GPE activated NRF2-dependent redox signaling, as evidenced by increased NRF2 nuclear translocation and transcriptional activity. Accordingly, GPE treatment significantly upregulated, or consistently increased, the expression of multiple NRF2 target genes, including HO-1, GPX, CAT, GCLC, and NQO1. Importantly, pharmacological inhibition of NRF2 attenuated GPE-induced ALP activity, confirming NRF2 as a key mediator of its osteogenic effects. Overall, grape pomace-derived polyphenols act as upstream modulators of redox-sensitive and osteogenic transcription factors, rebalancing MSC differentiation toward osteogenesis and mitigating age-related bone fragility. Full article

Smooth muscle (SM) dysfunction contributes to several pathological conditions, including atherosclerosis; current treatment strategies often fail to restore functional contractility. Adipose-derived stem cells (ADSCs) offer a promising cell source for regenerative medicine due to their accessibility and multipotency. Their differentiation into smooth muscle cells (SMC) is commonly driven by biochemical cues such as retinoic acid and transforming growth factor β; however, supporting this process with additional, non-invasive stimuli may enhance outcomes. Photobiomodulation (PBM) has emerged as a potential modulator of cellular metabolism, mitochondrial function and lineage commitment; however, its role in ADSCs to SMC differentiation remains insufficiently defined. ADSCs were irradiated with green (525 nm), near-infrared (825 nm) or dual wavelengths at 5 J/cm² and 10 J/cm² alongside the growth factors. Proliferation, cytotoxicity, mitochondrial membrane potential, collagen production, migration and smooth muscle marker expression were assessed. PBM induced a fluence-dependent biphasic response. 5 J/cm² fluences enhanced proliferation, mitochondrial activity, collagen deposition and organized SMC marker expression, whereas 10 J/cm² fluences lowered proliferation and membrane potential, reduced collagen and increased migration. PBM at 5 J/cm², especially greenlight, most effectively promoted ADSCs’ progression towards a SMC-like phenotype, with features consistent with a more contractile-like state. Full article

Mesenchymal stromal/stem cells (MSCs) are increasingly explored for immune-mediated diseases, yet standardized analytical readouts that capture coordinated immunomodulatory output across complementary secretory pathways remain limited. Here, we report the feasibility of an HPLC-based multi-analyte secretome characterization panel that quantifies two small-molecule outputs—adenosine and kynurenine—alongside two immunomodulatory proteins—interleukin-10 (IL-10) and transforming growth factor-beta (TGF-β)—in conditioned media from canine adipose-derived MSCs (cAD-MSCs). Canine immune-mediated hemolytic anemia (IMHA) was used as a disease context to motivate the selection of these analytes, given the pro-inflammatory cytokine environment characteristic of this condition. Three independent cAD-MSC lines were evaluated under baseline conditions and following cytokine stimulation with recombinant interferon-gamma (IFN-γ; 100 ng/mL) and tumor necrosis factor-alpha (TNF-α; 50 ng/mL), referred to herein as inflammatory priming or licensing. Conditioned media were collected at 72 h for metabolite analysis and 48 h for protein analysis, and quantified by HPLC using external calibration and peak integration. Across all three lines, licensing produced directionally consistent increases: mean adenosine increased 2.3-fold, mean kynurenine increased 3.1-fold, mean IL-10 increased 1.6-fold, and mean TGF-β increased 1.7-fold compared with unlicensed controls. Metabolite measurements for adenosine and kynurenine are reported with full chromatographic selectivity data; IL-10 and TGF-β measurements by reversed-phase HPLC with UV detection are presented as exploratory/semi-quantitative outputs and will require orthogonal confirmation (e.g., immunoassay) in future work. These findings are preliminary, derived from three independent donor lines with no comparator group, and are intended to support feasibility of the analytical framework rather than establish definitive performance specifications. Collectively, the data support the potential of a multi-analyte HPLC-based characterization panel to capture licensing-responsive secretory shifts across mechanistically complementary pathways, providing a foundation for expanded development and validation. Full article

Protease-activated receptor-1 (PAR-1) is a key regulator of mesenchymal stem cell (MSC) migration and tissue integration. Dabigatran, a direct thrombin inhibitor widely used as a non-vitamin K oral anticoagulant (NOAC), may affect PAR-1-mediated signaling pathways. This study investigated the effects of dabigatran on cell viability, apoptosis, and PAR-1 activity in adipose-derived MSCs (ADMSCs) in vitro. ADMSCs were exposed to five concentrations of dabigatran etexilate with thrombin activation. Cell viability was assessed using the MTT assay, apoptosis and morphological changes were evaluated via acridine orange/propidium iodide staining, and PAR-1 expression was analyzed by immunofluorescence. Results showed that high dabigatran concentration significantly reduced cell viability and induced apoptotic morphological changes. In contrast, lower, non-cytotoxic concentrations preserved normal fibroblastic morphology and maintained cell viability while reducing PAR-1 surface expression compared with thrombin-activated controls. These findings indicate that dabigatran at non-cytotoxic doses can modulate PAR-1 activity without compromising ADMSC survival. In conclusion, dabigatran influences MSC-related cellular functions beyond its anticoagulant properties. Full article

Ferroptosis of mesenchymal stem cells (MSCs) is a critical bottleneck restricting the efficiency of ruminant biological breeding. Phloretin, a natural bioactive polyphenol, exhibits potential ferroptosis-inhibitory activity. However, the regulatory effects and underlying mechanisms of phloretin on ruminant MSCs remain poorly understood. This study aimed to investigate the effects of phloretin on ferroptosis and elucidate its underlying molecular mechanisms. Herein, we isolated and cultured adipose-derived mesenchymal stem cells (AD-MSCs) from adipose tissue of a 9-day-old Leizhou goat and established a ferroptosis model in these cells using RSL3. We detected cell viability, proliferation, migration, ferroptosis-related indexes and key protein expression. The results showed that phloretin (25 and 50 μM) dose-dependently inhibited ferroptosis in goat AD-MSCs, reducing intracellular ferrous ion (Fe²⁺), reactive oxygen species (ROS) and lipid peroxidation levels, restoring glutathione content, and ameliorating mitochondrial structural damage. Mechanistically, phloretin exerted its anti-ferroptosis effects through direct antioxidant activity, activation of the Nrf2/HO-1/GPX4 signaling pathway and Fe²⁺ chelation. Nrf2 and GPX4 were key targets in this process. These results provide preliminary in vitro evidence and a theoretical basis for the potential application of phloretin in future research related to meat goat production and ruminant breeding. Full article

Human adipose biology is strongly influenced by three-dimensional (3D) architecture, cell–cell interactions, and local oxygen availability maintained over a long-term culture period, features that are not reproduced in traditional two-dimensional (2D) culture systems. To address this gap, we established a long-term human adipose-derived stem cell (hASC) spheroid model using elastin-like polypeptide–polyethyleneimine (ELP-PEI) coating. The ELP-PEI coating facilitated stable spheroid formation and sustained adipogenic differentiation over 56 days. As spheroids enlarged and matured, they exhibited hallmark features of adipocytes, including lipid accumulation, morphological compaction, and transition out of the proliferative state. Glucose uptake increased during maturation and declined as spheroids became larger. This reduction coincided with a marked rise in hypoxia-inducible factor-1α (HIF-1α) expression, indicating the emergence of a hypoxic microenvironment within larger spheroids. Notably, inhibiting HIF-1α restored insulin-stimulated glucose uptake, demonstrating that hypoxia was the primary driver of impaired insulin responsiveness in late-stage spheroids. These findings position ELP-PEI-supported hASC spheroids as a practical and physiologically relevant platform for studying human adipocyte biology, particularly the development and reversibility of hypoxia-associated metabolic dysfunction. This model offers new opportunities for mechanistic studies and for evaluating therapeutic strategies targeting insulin resistance and adipose tissue pathology. Full article

Bone regeneration requires tight coordination between mesenchymal stem cells (MSCs), immune signaling, and extracellular matrix remodeling. Yet, how atypical immune receptors contribute to this process remains unclear. Here, we identify Toll-like receptor 10 (TLR10) as a key regulator of osteogenic differentiation in human adipose-derived MSCs. Herein, ASC/TERT1 MSCs were engineered to overexpress or silence TLR10 using lentiviral vectors, and osteogenic differentiation (0–14 days) was assessed by metabolic assays—RT-qPCR of COL1A2, ALPL and BGLAP—Alizarin Red S staining, and quantitative mass spectrometry. Enhancing TLR10 expression promoted osteogenic gene programs, extracellular matrix organization, metabolic adaptation, and robust matrix mineralization, whereas TLR10 suppression maintained proliferative states and impaired osteoblast maturation. Proteomic analyses revealed that TLR10 selectively activates osteogenic, ECM-remodeling, and vitamin D-responsive pathways, while restraining programs antagonistic to differentiation. Notably, active vitamin D induced TLR10 expression and partially restored osteogenesis in TLR10-deficient cells, indicating that TLR10 is associated with vitamin D-driven bone formation. Together, beyond its established role in innate immunity, TLR10 emerges as a vitamin D-responsive regulator of mesenchymal stem cell osteogenesis, highlighting a potential therapeutic axis to enhance bone regeneration and osteogenic outcomes. Full article

Glucagon-like peptide-1 (GLP-1) is an incretin hormone involved in glucose regulation. Glucagon-like peptide-1 receptor agonists (GLP-1 RAs) are widely used in the treatment of type 2 diabetes mellitus and obesity, as well as in cardiovascular risk reduction. Recent evidence suggests that GLP-1 receptors are expressed in cutaneous tissues; however, their role in skin homeostasis and aging remains insufficiently clarified. This review summarizes recent experimental and clinical studies examining the effects of GLP-1 receptor agonists on skin homeostasis, wound healing, regeneration, and aging processes. Experimental data indicate that GLP-1 RAs may promote wound healing through modulation of inflammatory pathways, enhancement of keratinocyte migration, improved microvascular perfusion, and support of fibroblast function. Antioxidant and cytoprotective mechanisms have also been described. Conversely, rapid weight loss associated with GLP-1 RA therapy has been linked to structural facial changes, including reduction in dermal white adipose tissue and decreased collagen synthesis, which may clinically resemble accelerated skin aging. Mechanistic findings suggest heterogeneous and pathway-dependent effects. Overall, the impact of GLP-1 receptor agonists on skin biology appears multifaceted, and further well-designed clinical studies are required to determine their precise dermatological implications. Full article