Search Results (1,685)

The response of breast cancer to chemotherapy is strongly influenced by tumor–microenvironment interactions and the presence of cancer stem-like cells (CSCs), which are associated with therapy resistance. The aim of this study was to investigate whether modulation of adipocyte–tumor crosstalk by licorice root extract (LRE) could influence doxorubicin response in CSC-enriched MCF-7 mammospheres. MCF-7 cells were pretreated with conditioned media derived from preadipocytes, mature adipocytes, or LRE-treated mature adipocytes, or were directly exposed to LRE, and subsequently cultured as mammospheres. CSC enrichment was supported by increased expression of CD44 and CD133. Mature adipocyte-conditioned medium promoted the formation of larger and more compact spheroids, reduced intracellular reactive oxygen species (ROS) and apoptosis, and decreased doxorubicin uptake. In contrast, conditioned medium from LRE-treated adipocytes partially counteracted these effects by increasing oxidative stress, apoptotic response, and intracellular doxorubicin accumulation. Collectively, these findings support the involvement of adipocyte–tumor crosstalk in modulating functional parameters associated with doxorubicin responsiveness and suggest that modulation of the adipose microenvironment by LRE may represent a promising area for further investigation in the context of microenvironment-driven chemoresistance. Full article

Keloids are characterized by fibrosis and chronic inflammation, but links between mitochondrial dysfunction and keloid pathogenesis remain unclear. This study examined whether impaired PINK1/Parkin-dependent mitophagy is associated with mitochondrial DNA (mtDNA)-mediated innate immune activation and fibrosis in keloids, and evaluated mitochondrial transplantation as a potential therapeutic strategy. Primary keloid fibroblasts (KFs), normal skin fibroblasts (NFs), adipose-derived stem cells (ADSCs), human keloid tissues, and human keloid xenografts in immunodeficient BALB/c nude mice were analyzed using ultrastructural, molecular, and functional approaches. Freshly isolated NF-derived mitochondria (nMito) and ADSC-derived mitochondria (aMito) were compared at protein-equivalent doses. KFs exhibited mitochondrial abnormalities, impaired oxidative phosphorylation, increased reactive oxygen species, mtDNA leakage, and cGAS/STING pathway activation. Elevated PINK1 expression, reduced Parkin expression and p62 accumulation were consistent with impaired downstream mitophagic clearance. Both nMito and aMito were associated with improved mitochondrial function, changes in mitophagy-related markers, reduced cytosolic mtDNA and cGAS/STING signaling, and attenuated fibroblast activation, with greater aMito-associated changes in selected endpoints. In xenografts, intralesional administration of either mitochondria improved collagen organization and reduced fibrotic and inflammatory signaling. Together, these findings link altered PINK1/Parkin-dependent mitophagy to mtDNA-driven inflammation and fibrosis and support mitochondrial transplantation as a potential organelle-based therapeutic approach. Full article

Feline mesenchymal stem cells (MSCs) isolated from adipose (A-MSCs) and uterine (U-MSCs) tissues during routine ovariohysterectomy represent valuable therapeutic sources in veterinary regenerative medicine. While quantitative real-time PCR (qRT-PCR) remains the gold standard for transcriptional profiling in these cells, its analytical reliability relies on empirically validated reference genes. In this study, we assessed the expression stability of nine candidate reference genes (ACTB, B2M, GAPDH, GUSB, HMBS, HPRT1, RPL7, TBP, and YWHAZ) in feline A-MSCs and U-MSCs using the geNorm and NormFinder algorithms. Both computational approaches identified TBP and HMBS as the most stably expressed genes across both cell types, whereas the historically popular GAPDH was the least stable. All geNorm pairwise variation values were below the commonly used threshold of 0.15 in both A-MSCs and U-MSCs. To illustrate the impact of reference gene choice, we evaluated the pluripotency marker OCT4. Normalizing using TBP and HMBS revealed significant differences in OCT4 expression between A-MSCs and U-MSCs (p < 0.05). In contrast, applying the unstable GAPDH masked this difference. These findings identify TBP and HMBS as promising reference gene candidates for qRT-PCR normalization in P3 undifferentiated feline A-MSCs and U-MSCs. Full article

Peripheral neuropathy (PN) is a debilitating condition characterized by chronic pain, numbness, and motor dysfunction, with limited treatment options. Ischemic stroke can cause central neuropathy, which may also induce PN. Human mesenchymal stem cells (hMSCs) have shown promise in therapeutic applications, but limitations in cell viability, immune response, and efficacy persist. Extracellular vesicles (EVs), which facilitate cell-free intercellular communication, offer a promising alternative for nerve regeneration. Electrical stimulation (ES) has emerged as a method to enhance EV secretion, and this study investigates its potential for promoting EV production from human adipose tissue-derived mesenchymal stem cells (hASCs) and human Schwann cells (hSCs). In this study, hASCs, hSCs, and lipopolysaccharide (LPS)-induced inflamed hSCs were subjected to one hour of low-frequency direct current (DC) electrical stimulation (100 mV/mL) for 7 days. EVs were isolated using differential ultracentrifugation and characterized through nanoparticle tracking analysis (NTA). Gene expression was analyzed via qRT-PCR to evaluate markers associated with EV biogenesis as well as pro- and anti-inflammatory cytokines. Our results demonstrate that ES significantly increases EV secretion from both hASCs and hSCs, with a notable upregulation of genes involved in both the endosomal sorting complex required for transport (ESCRT)-dependent and ESCRT-independent pathways of EV biogenesis. Additionally, ES modulates inflammation-related markers, promoting anti-inflammatory gene expression and reducing pro-inflammatory gene levels. Notably, LPS-induced hSCs exhibited a phenotype shift from myelinating to non-myelinating cells, producing EVs capable of modulating the inflammatory microenvironment. However, prolonged exposure to ES led to a decrease in EV secretion and changes in EV size distribution, suggesting potential cellular adaptation or membrane stress. This study highlights the potential of ES as a scalable, cell-free strategy to enhance EV production, offering new insights into its therapeutic applications for peripheral neuropathy and nerve regeneration. Full article

Background/Objective: Liver cancer is a global health challenge due to its resistance to most systemic therapies. Human mesenchymal and epithelial stem cells exhibit anti-proliferative and pro-apoptotic effects on some cancer cell lines, which can be used to support standard therapy. The aim of this study was to investigate the effects of conditioned media (CM) derived from mesenchymal adipose tissue-derived stem cells (hADSCs) and amniotic membrane-derived cells expressing both mesenchymal and epithelial markers (hACs) on HepG2 liver cancer cells in vitro and to identify possible similarities to the action of sorafenib, a standard drug in hepatocellular carcinoma (HCC) therapy. Methods: HepG2 cells were cultured with hADSC-derived CM (CM-hADSC) or hAC-derived CM (CM-hAC), either alone or with 7.5 μm sorafenib for 48 h. HepG2 cell viability and the expression of genes and/or proteins related to the apoptosis, cell-cycle, and Ras/Raf/MEK/ERK signaling pathway, were assessed. Results: The effect of sorafenib administration alone, consisting in reducing HepG2 cell viability, cell cycle inhibition, and reducing the expression of the alpha-fetoprotein (AFP) gene and most proteins related to the Ras/Raf/MEK/ERK pathway, was dominant over the effect of CM administered in combination with one of them. The effect of sorafenib on increasing the number of early apoptotic cells was not associated with increased Bax protein expression or with an increased proportion of cleaved forms of caspase-7 and caspase-9. The effect of CM-hAC was often different from that of CM-hADSC and consisted in enhancing the inhibitory effect of sorafenib at the cell cycle level and increasing the number of apoptotic cells and caspase mRNA expression, while activating the Ras/Raf/MEK/ERK signaling pathway and inducing AFP gene expression. Conclusions: Combined administration of sorafenib and some CM may produce the desired effects by enhancing the action of the drug. However, some of the effects of sorafenib and CM derived from different stem cell types were significantly different, which means that the role of stem cell secretome as a potential anticancer therapy requires further investigation, taking into account different types of cancer cell lines and a thorough analysis of the secretome composition. Full article

Estrogen deficiency induces bone loss and is accompanied by lipid metabolic disturbances. This study investigated the protective effects of Larimichthys crocea-derived calcium-chelating peptides (LCP) in a leuprorelin-induced estrogen-deficient mouse model. LCP attenuated tibial trabecular deterioration, increased serum procollagen type I N-terminal propeptide (PINP), osteocalcin (OCN), and bone-specific alkaline phosphatase (BALP), and reduced C-terminal telopeptide of type I collagen (CTX-I). Histological analyses further showed increased trabecular area and OCN expression and reduced marrow adiposity. LCP also attenuated alterations in serum lipid profiles and circulating levels of leptin and adiponectin, while reducing adipogenesis-related gene expression in femoral tissue and white adipose tissue. Exploratory assays using group-pooled cultures of bone marrow mesenchymal stem cells (BMSCs) showed differentiation-related patterns that were directionally consistent with the in vivo findings, but these descriptive observations require confirmation using independent biological replicates. Correlation analysis revealed associations among bone mass, osteogenesis-related indicators, and lipid metabolism-related parameters. Collectively, LCP attenuated estrogen deficiency-induced bone loss together with favorable changes in indicators related to bone formation, adipogenesis, and lipid metabolism. These findings support further investigation of LCP as a food-derived nutritional ingredient for supporting bone health under estrogen-deficient conditions. Full article

Magnetic Particle Imaging (MPI) detects superparamagnetic nanoparticles, enabling bimodal contrast with MRI. Resovist^®/Ferucarbotran, still used in research, has clinical safety compatibility but suboptimal MPI performance due to many small 5 nm SPIO cores. Magnetic fractionation can enrich larger cores, improving MPI signal and supporting cellular imaging applications. To enable bimodal MRI/MPI and assess in vivo extracellular vesicle (EV) labelling, we characterized the imaging sensitivity and cell-labelling performance of VivoTrax, a commercial formulation similar to Resovist^®, and VivoTrax Plus, obtained by magnetic fractionation. VivoTrax Plus showed higher MRI transverse relaxivity and MPI sensitivity than VivoTrax, and both formulations displayed low toxicity toward adipose-derived stem cells (ASCs). VivoTrax Plus allowed MRI detection of small cell numbers, around 100 cells, in agarose phantoms with greater sensitivity than VivoTrax. However, EVs of 30–150 nm isolated from ASCs labelled with VivoTrax Plus did not retain SPIONs, whereas EVs from VivoTrax-labelled ASCs did. Preliminary in vivo experiments in SOD-G93A mice showed MRI-detectable signal voids in the brain after intranasal EV administration, suggesting EV migration to lesioned areas. Overall, magnetic fractionation improves SPION imaging sensitivity but may alter relevant biological properties. Full article

Tendinopathies represent a major clinical challenge. Vasculature, neuromuscular junctions, low cellularity, and slow extracellular matrix (ECM) turnover restrict endogenous repair and predispose injured tendons to fibrosis, mechanical weakness, and reinjury. Current therapeutic strategies including rehabilitation protocols, anti-inflammatory medications, platelet-rich plasma (PRP) injections, and surgical repair primarily address symptoms or structural deficits without correcting the underlying biological limitations of tendon healing. Cell-based therapies have emerged as a promising regenerative approach aimed at restoring tissue homeostasis through modulation of angiogenesis, collagen synthesis, immune responses, and tenogenic differentiation. Mesenchymal stem cells (MSCs), adipose-derived stem cells (ADSCs), tendon-derived stem cells (TDSCs), induced pluripotent stem cells (iPSCs), differentiated tenocytes, and extracellular vesicle (EV)-based products have demonstrated the ability to enhance vascularization, promote type I collagen remodeling, suppress excessive inflammation, and stimulate tenocyte lineage commitment. These effects are mediated through paracrine signaling, growth factor secretion, and activation of key pathways, including HIF-1α, TGF-β/SMAD, NF-κB, and PI3K/Akt signaling. Despite promising preclinical data, significant translational challenges remain, including limited cell survival at the injury site, variability in cell sources and dosing, immunogenicity, risk of misdifferentiation, and lack of standardization across clinical protocols. Emerging strategies such as genetic modification, hypoxic preconditioning, scaffold-based delivery systems, and extracellular vesicle engineering aim to enhance therapeutic efficacy and reproducibility. This review synthesizes current evidence on cell-based tendon repair, critically evaluates mechanistic insights, clinical trials, and translational barriers, and outlines future directions toward biologically informed regenerative therapies. Full article

Advancing age is often accompanied by a gradual decline in musculoskeletal (MSK) health and integrity, making the skeleton not just a structural framework; but a dynamic indicator of biological ageing. Bone tissue undergoes continuous remodelling through tightly regulated interactions among osteocytes, osteoblasts, osteoclasts, progenitor cells or mesenchymal stem/stromal cells (MSCs) and growth factors. With advancing age, this balance shifts toward increased resorption, increased adipose tissue formation, reduced bone formation, and deterioration of bone microarchitecture. This eventually can lead to frailty, increased fracture risk and other age-related diseases (ARDs) due to ageing, cellular senescence and inflammaging. Interestingly, bone-derived changes often precede clinical manifestations of these ARDs, suggesting its potential use as an early biomarker for biological ageing and longevity risks. In our review, we explore the possibility of using the bone as a biomarker for ageing, followed by anti-ageing strategies to encourage bone longevity. We begin by discussing our current knowledge of the bone, the bone marrow (BM), changes in the BM with ageing and current methods of tracking bone ageing; both clinically and experimentally. We then explore studies that investigated the anti-ageing strategies, specifically focused on diet, exercises and lifestyle to shift towards enhanced bone health and longevity. Finally, we discuss the limitations of achieving this, what the future of this field may look like and what methods may be used to translate the idea of bone as a biomarker of ageing, from bench to bed. Full article

Perirenal adipose tissue (PRAT) has emerged as a clinically relevant endocrine organ connecting obesity to cardiovascular disease (CVD), chronic kidney disease, and certain malignancies. Its unique anatomical location, surrounding the kidneys, accounts for PRAT’s role in altering intrarenal haemodynamics and hydrostatic pressure. Accordingly, PRAT’s expansion is associated with the activation of the renin–angiotensin–aldosterone system (RAAS), further increasing blood pressure. Adipokine dysregulation, together with overexpression of miR-24-3p, miR-155, miR-146a, and miR-21 in PRAT, modulates inflammation and oxidative stress, leading to endothelial dysfunction and increased risk of atherosclerosis and hypertension in obesity. Imaging assessment of PRAT thickness through computed tomography, magnetic resonance, or ultrasound has also emerged as a complementary measure for the evaluation of CVD risk. Potential therapeutic strategies targeting PRAT include lifestyle interventions, antidiabetic agents, RAAS inhibitors, adipose tissue browning agents, NOD-like receptor protein 3 (NLRP3) inflammasome inhibitors, peroxisome proliferator-activated receptor gamma (PPARγ) agonists, and surgery. Currently, novel therapeutic interventions targeting PRAT activity in CVD, such as senotherapeutic strategies, bioengineering approaches aimed at enhancing adipose-derived mesenchymal stem cell (ADMSC) function, gut microbiota modulation, and colchicine and bone morphogenetic protein 4 (BMP4) administration, are also being explored. In light of these findings, PRAT’s clinical relevance extends beyond its energy storage role, highlighting it as a metabolically active fat depot. Its assessment and therapeutic modulation may complement existing cardiovascular prevention strategies, particularly in patients with obesity. Full article

Oocyte quality is the primary determinant of success in assisted reproductive technologies (ART), and mitochondrial dysfunction is increasingly recognized as a central mediator of poor oocyte competence across advanced maternal age, recurrent implantation failure, polycystic ovary syndrome, endometriosis, and obesity. Chemical interventions improve the mitochondrial microenvironment but cannot restore depleted mitochondrial mass, while heterologous mitochondrial replacement remains constrained by ethical, legal, and biological limitations. This review examines the biological basis for mitochondrial intervention in oocytes, evaluates chemical and cellular therapeutic approaches, and assesses the evidence for autologous Adipose Stem Cell-derived Mitochondria ENergy Transfer (ASCENT). Mitochondria govern oocyte ATP production, calcium-mediated meiotic integrity, and redox homeostasis, and their disruption contributes to aneuploidy, fertilization failure, and embryonic arrest. Among cellular interventions, autologous adipose-derived stem cell mitochondrial transplantation offers minimally invasive tissue accessibility, morphological compatibility with oocyte mitochondria, robust membrane potential, and a preclinically validated Mito-ICSI delivery platform. Notably, ASCENT is currently the only autologous approach for which safety across three consecutive offspring generations has been reported in a mammalian model, with primary maternal origin of offspring mtDNA confirmed. Together, preclinical efficacy, transgenerational safety, and human proof-of-concept support progression toward a rigorously designed clinical trial, while ASC-derived mitochondria hold broader relevance in regenerative medicine. Full article

Objective: To evaluate whether human adipose-derived mesenchymal stem cells (ADSCs) protect against ᴅ-galactose (ᴅ-gal)-induced aging-like testicular injury and to investigate the involvement of the Keap1/Nrf2 pathway and NLRP3-associated pyroptosis. Methods: A mouse model of aging-like testicular injury was established by subcutaneous administration of ᴅ-gal for 8 weeks, followed by tail vein injection of ADSCs. Testicular morphology, blood–testis barrier (BTB) integrity, and senescence-associated markers (p16, p21) were assessed. In vitro, TM4 Sertoli cells were used to establish a senescence model and Tranwell co-cultured with ADSCs. Oxidative stress, inflammatory responses, and pyroptosis-related markers were evaluated using biochemical assays, immunofluorescence, Western blotting, and RT-qPCR. The involvement of the Keap1/Nrf2-NLRP3 axis was further examined using pharmacological inhibitors. Results: ADSC treatment significantly alleviated ᴅ-gal-induced testicular atrophy and histopathological injury, accompanied by reduced expression of the senescence markers p16 and p21 and partial restoration of BTB-related structures. ADSCs also attenuated oxidative stress, as evidenced by decreased ROS and MDA levels, increased SOD activity, and enhanced expression of Nrf2 and its downstream antioxidant targets, including HO-1 and NQO1. In parallel, ADSC administration suppressed NLRP3 activation, reduced caspase-1 cleavage, and lowered the expression of pro-inflammatory cytokines. In TM4 cells, inhibition of Nrf2 weakened the protective effects of ADSCs and was accompanied by reactivation of NLRP3-associated signaling, whereas inhibition of NLRP3 attenuated senescence- and inflammation-related changes without restoring Nrf2 activity. Conclusions: ADSCs alleviate ᴅ-gal-induced aging-like testicular injury, at least in part, by restoring redox balance, preserving BTB-associated structure, and suppressing NLRP3-associated pyroptosis through the Keap1/Nrf2 pathway. These findings suggest that ADSC-based therapy may represent a promising strategy for age-related male reproductive dysfunction, while further studies using genetic models and functional fertility endpoints are needed to confirm causality and translational relevance. Full article

Background: Secondary lymphedema is a chronic complication of oncologic surgery and radiotherapy for which effective disease-modifying therapies remain lacking. While mesenchymal stem cells (MSCs) have shown partial benefit in experimental models, direct functional comparison with lineage-committed lymphatic endothelial progenitor cells (LEPCs) remains limited. Methods: Secondary lymphedema was induced in C57BL/6J mice by circumferential tail skin excision with disruption of superficial lymphatics. Animals received intradermal phosphate-buffered saline (PBS), MSCs, or adipose-derived LEPCs on postoperative days 1 and 7. Lymphatic function was longitudinally quantified using IVIS-based near-infrared indocyanine green (ICG) imaging with standardized region-of-interest analysis. Tail diameter was measured serially throughout follow-up as a complementary morphometric parameter. Tissue remodeling was assessed by Picrosirius Red staining and qualitative LYVE-1 immunofluorescence. Statistical analysis incorporated mixed-effects modeling to evaluate treatment group, sex, time, and their interaction terms, with estimation of effect sizes and 95% confidence intervals. Results: LEPC-treated mice demonstrated higher IVIS signal intensity than MSC- and PBS-treated animals, consistent with improved lymphatic transport at the injury site. Longitudinal tail diameter analysis showed a more favorable temporal profile in the LEPC group relative to controls, with exploratory analysis suggesting potential variations in temporal profiles between sexes that warrant further investigation in larger cohorts. Histologic analysis showed reduced non-tissue/void area and a more favorable qualitative pattern of LYVE-1 staining in LEPC-treated tissue. Conclusions: LEPC administration enhanced functional and structural recovery in a murine model of secondary lymphedema. These findings support further translational evaluation of LEPC-based approaches as a regenerative strategy for secondary lymphedema. Full article

Background and Objectives: Autologous fat transplantation using fat alone can lead to postoperative complications. Laminin–alginate beads have been investigated as carriers for adipose mesenchymal stem cells (ASCs), with the potential to modulate cell release kinetics, influence macrophage responses, and thereby affect the maintenance of graft volume. We hypothesized that encapsulating ASCs in laminin–alginate beads would allow gradual ASC release into the transplantation site and influence the macrophage-related microenvironment during fat graft maturation. Materials and Methods: ASCs were isolated from two female enhanced green fluorescent protein (eGFP) transgenic rats. For SC (ASC + fat) grafts, ASCs were freely mixed with 1 mL of mature adipose tissue. For bead-encapsulated cell (BEC) (laminin–alginate + ASC + fat) grafts, ASCs were first encapsulated in laminin–alginate beads and subsequently mixed with 1 mL of fat tissue. The fat graft was subcutaneously injected into the backs of the rats, which were subsequently euthanized at weeks 4, 6, and 8. The grafts were then harvested for histological and immunofluorescence analyses. Results: The histological analysis indicated that the number of eGFP-ASCs in the BEC grafts gradually increased between weeks 4 and 8, whereas that in the SC grafts decreased over the same period. The BEC grafts also exhibited a numerically higher degree of M2 macrophage infiltration. Conclusions: Laminin–alginate bead encapsulation was associated with prolonged persistence of eGFP-ASCs within the graft area. Although the macrophage-related findings were not statistically significant, they suggest a potential interaction between gradual ASC release and the local graft microenvironment. Full article

Urine-derived stem cells (UDSCs) have emerged as a promising cell source for regenerative medicine due to their non-invasive procurement, high proliferative capacity, and potential relevance to kidney-specific repair. Unlike conventional mesenchymal stem cells (MSCs) obtained from bone marrow or adipose tissue, UDSCs originate from multiple regions of the urinary tract and exhibit a unique biological profile that combines MSC characteristics with features of renal progenitor populations. This review provides a comprehensive overview of the current understanding of UDSC biology, including their origin, isolation strategies, morphology, immunophenotypic characteristics, differentiation potential, and secretory profile. Particular attention is given to the expression of renal lineage-associated markers and pluripotency-related factors that may contribute to their regenerative capacity. The bioactive mediators of UDSCs regulate inflammation, oxidative stress, angiogenesis, and extracellular matrix remodeling, thereby influencing key pathways implicated in chronic kidney disease (CKD)-associated fibrosis. Furthermore, the intrinsic renal progenitor signature of UDSCs may provide advantages in renal homing and tissue-specific repair compared with conventional MSC populations. Despite encouraging preclinical findings, significant challenges remain, including cellular heterogeneity, inconsistent isolation efficiency, lack of standardized characterization criteria, and limited clinical validation. Collectively, current evidence positions UDSCs as a biologically distinct and therapeutically attractive platform for kidney regeneration. Full article