Introduction: Mesenchymal stem cells (MSCs) play a significant role in shaping and regulating microenvironments. The factors influencing the glycolytic metabolic reprogramming of MSCs remain unknown. Methods: MSCs were isolated from human umbilical cords and identified using flow cytometry. Glycolysis-related gene expression, glucose consumption, and ATP and lactate production were measured in MSCs with HIF1A-AS3 overexpression or knockdown. The roles of HIF1A-AS3 in the glycolytic pathway and its downstream network were revealed by high-throughput sequencing. Results: HIF1A-AS3 enhanced the glycolytic level of MSCs by increasing glucose consumption, lactate production, ATP levels, and expression of key glycolytic enzymes (GLUT1, HK2, and PKM2). The downstream signaling pathway of HIF1A-AS3 might be associated with glycosylationrelated diseases. Discussion: It was demonstrated that HIF1A-AS3, a hypoxia-induced long non‑coding RNA, acted as a critical regulator of glycolytic reprogramming in MSCs. Additionally, HIF1A-AS3 upregulated a distinct subset of interferon‑stimulated genes, suggesting a potential link between metabolic reprogramming and immunomodulation in MSCs. Further mechanistic studies are warranted to elucidate how HIF1A-AS3 regulates these processes. Conclusion: The study reveals that HIF1A-AS3 is a regulator of the metabolic reprogramming of glycolysis in MSCs. These findings provide new insights into understanding the metabolic regulatory mechanism of MSCs.
Introduction/ Objective: Neutrophils are crucial for an effective innate immune response. However, overactivation is associated with chronic inflammation, as in systemic lupus erythematosus (SLE). Gingival mesenchymal stromal cells (GMSCs) are used as anti-inflammatory therapies because they possess strong immunomodulatory properties, primarily acting through paracrine mechanisms. GMSCs exhibit immunoplasticity in response to their microenvironment and adjust their secretion profile. This study aimed to investigate the effects of conditioned media derived from human gingival mesenchymal stromal cells (hGMSC-CM) or from cells pretreated with IFNγ (hGMSCγ- CM) on neutrophil antimicrobial and proinflammatory functions. METHODS:hGMSCs were isolated to meet the criteria of the International Society for Cell & Gene Therapy (ISCT). Neutrophil intracellular reactive oxygen species (ROS) production; neutrophil elastase (NE), cathepsin G (CG), and myeloperoxidase (MPO) enzymatic activities; induction of lytic and vital neutrophil extracellular traps (NETs); and bacterial uptake and engulfment were analyzed using flow cytometry, photometric methods, and fluorescence and confocal microscopy. RESULTS:Both hGMSC-derived CMs significantly reduced intracellular ROS levels, reduced the enzymatic activities of MPO, CG, and NE by decreasing degranulation, and diminished NET formation (DNA, LL37, and PAD4) through both the lytic and vital pathways, including NET production observed in an inflammatory environment using plasma from SLE patients. DISCUSSION:These inhibitory effects were more pronounced with hGMSCγ-CM. Interestingly, hGMSCγ-CM did not impair bacterial uptake, engulfment, or neutrophil microbicidal activity against Staphylococcus aureus and Escherichia coli, thereby maintaining the functional balance of neutrophils. CONCLUSION:These findings support the use of hGMSCγ-CM as a cell-free biological product for therapies targeting inflammation driven by neutrophil overactivation.
An abdominal aortic aneurysm (AAA) is a serious vascular disease characterized by progressive dilation of the abdominal aorta, resulting in an increased risk of rupture. AAA develops through multiple pathogenic mechanisms, including chronic inflammation, extracellular matrix (ECM) degradation, oxidative stress, and vascular smooth muscle cell (VSMC) apoptosis. Surgical intervention is the primary treatment for AAA; however, no effective pharmacological therapies are currently available to halt or reverse disease progression. This review evaluates the therapeutic potential of multipotent mesenchymal stromal/stem cells (MSCs) for AAA, with particular emphasis on their immunomodulatory and regenerative properties. Preclinical studies suggest that MSCs can attenuate AAA progression in animal models by modulating the inflammatory microenvironment, reducing matrix metalloproteinase activity, and promoting tissue repair. In addition, interactions between MSCs and various immune cells decrease inflammatory responses, thereby positively impacting the structure of the aortic wall. Collectively, the available evidence highlights the broad therapeutic potential of MSCs in modulating key molecular and cellular pathways involved in AAA progression. Given their ability to modulate the immune system and promote vascular regeneration, MSCs may have significant potential for developing disease-modifying therapies for AAA. By targeting key pathogenic processes involved in AAA progression, MSCs offer the potential to complement existing surgical management and address mechanisms that are not targeted by current therapies. However, additional well-designed laboratory-based and clinical studies are needed to optimize MSC-based therapeutic strategies and to establish their long-term efficacy and safety before clinical translation.
Introduction: Mesenchymal stem cells (MSCs) possess immunomodulatory and tissuereparative properties and have shown therapeutic potential in acute lung injury (ALI). However, the mechanisms underlying their protective effects remain unclear. This study aimed to evaluate the effects of MSCs on bleomycin-induced ALI and to explore the underlying molecular mechanisms. Methods: A bleomycin-induced ALI mouse model was established, followed by human umbilical cord-derived MSC administration. Lung injury, pulmonary edema, neutrophil infiltration, and apoptosis of alveolar type II epithelial cells (AT2) were assessed. RNA-seq and molecular validation were performed using sorted AT2 cells to investigate the underlying mechanism. In vitro assays were carried out to examine the role of Ntrk2 in MSC-mediated epithelial repair. Results: MSC treatment alleviated lung injury, improved alveolar structure, reduced pulmonary edema, and decreased neutrophil infiltration and AT2 cell apoptosis. RNA-seq and molecular analyses indicated that MSCs regulated Ntrk2 expression by suppressing NF-κB signaling and enhancing STAT3 signaling, thereby promoting alveolar epithelial regeneration. In vitro, inhibition of Ntrk2 impaired the promotive effects of MSCs on A549 cell migration and alveolar organoid repair. Discussion: These findings suggest that MSCs protect against ALI by attenuating inflammatory injury and promoting alveolar epithelial repair, with the NF-κB/STAT3-Ntrk2 axis contributing to this process. Conclusion: MSC treatment ameliorated bleomycin-induced ALI and promoted alveolar epithelial regeneration, at least partly through the NF-κB/STAT3-Ntrk2 axis. These results provide further mechanistic support for MSC-based therapy in ALI.
Introduction: The cyclical growth of hair follicles depends on the periodic proliferation and differentiation of hair follicle stem cells (HFSCs). Previous studies have shown that extracellular vesicles (EVs) derived from mesenchymal stem cells (MSCs) and human hair outer root sheath cells (HHORSCs) can enhance hair follicle development, ameliorate androgenetic alopecia, and support the inductive capacity of dermal papilla cells. However, the regulatory effects of HHORSC-derived EVs (HHORSC-EVs) and human bone marrow mesenchymal stem cell-derived EVs (HBMMSCEVs) on HFSCs remain unclear. Methods: HFSCs were treated with HHORSC-EVs or HBMMSC-EVs for 10 days. The effects of EVs on HFSC proliferation, apoptosis, and differentiation were evaluated using a Cell Counting Kit- 8 assay, flow cytometry, and immunofluorescence staining, respectively. Results: EVs from both HHORSCs and HBMMSCs did not enhance hair follicle growth by improving the proliferation of HFSCs, but rather by inhibiting apoptosis. In addition, HBMMSC-EVs further promoted HFSC stemness maintenance by upregulating K15 and CD34 expression. Discussion: HFSCs may serve as a novel target for both HHORSC-EVs and HBMMSC-EVs, highlighting their therapeutic potential for hair loss disorders. Conclusion: This study offers new insights into developing EV-based therapies for hair loss disorders.
BACKGROUND:This study aimed to investigate the therapeutic effect and potential mechanism of intravenous injection of human umbilical cord mesenchymal stem cells (hUCMSCs) on acute lung injury (ALI) in rats. METHODS:Wistar albino rats were used to establish an ALI animal model via intravenous oleic acid suspension injection. Rats in the low-dose and high-dose mesenchymal stem cell (MSC) groups were treated with 2.5×10^5 hUCMSCs and 5×10^5 hUCMSCs, respectively. After 24 hours of modelling, the rats were sacrificed and samples were collected. The lung coefficient was calculated, and lung tissue morphology was examined. The expression levels of TNF-α, IL-1β, IL-4, IL-6, and IL-10 in lung tissue and serum were assessed. Additionally, the expression of Claudin-5, Occludin, and ZO-1 in the lung was measured. RESULTS AND DISCUSSION:Compared with the control group, lung volume and lung coefficient significantly increased, and lung tissue exhibited pathological changes with an increased lung injury score. Levels of TNF-α, IL-1β, and IL-6 in lung tissue and serum significantly increased, while IL-4 and IL10 levels significantly decreased. The proportion of neutrophils markedly increased, and the expression of Claudin-5, Occludin, and ZO-1 in the lung significantly decreased in the model group. These results indicate that oleic acid can induce acute lung injury and activate inflammatory. Compared with the model group, both MSC-treated groups showed significant reductions in lung coefficient and lung injury score. Levels of TNF-α, IL-1β, and IL-6 in lung tissue and serum significantly decreased, while IL-4 and IL-10 levels significantly increased. Furthermore, the proportion of neutrophils decreased significantly, and the expression of Claudin-5, Occludin, and ZO-1 in the lung increased significantly in the two MSC-treated groups. The therapeutic effect can likely be ascribed to the immunomodulatory properties of hUCMSCs and capacity to mitigate barrier damage, as substantiated by the decreased levels of pro-inflammatory cytokines, along with the elevated expression of antiinflammatory factors and tight junction proteins in lung tissue. CONCLUSION:The hUCMSCs can effectively ameliorate oleic acid-induced ALI in rats by exerting anti-inflammatory effects and protecting the alveolar-capillary endothelial barrier.
INTRODUCTION:Pancreatic stellate cells (PSCs) play a central role in pancreatic physiology and disease, and the transition between their quiescent and activated states influences processes such as fibrosis and regeneration. However, the stem cell-like properties of PSCs remain unclear. This study aims to clarify the stem cell characteristics of PSCs and establish a method to maintain their quiescent state, thereby providing a new perspective on pancreatic regeneration. METHODS:PSCs from Lrat-cre; Rosa26-tdTomato mice were fate-traced, and immunofluorescence was used to assess Lrat, Oct4, and Nanog co-localization. Primary PSCs from C57BL/6 mice were cultured in mesenchymal stem cell (MSC) or DMEM/F12 medium. Lipid droplets, morphology, activation markers (α-SMA, collagen-I, fibronectin), and stemness-associated markers were analysed via qRT-PCR and immunofluorescence. RESULTS:Lrat co-localized with Oct4/Nanog in mice. MSC-cultured PSCs had more lipid droplets, a quiescent morphology, and lower levels of activation markers than DMEM/F12-cultured cells. They also expressed stemness-associated markers (Pdx1, Oct4, Nkx6.1, Ngn3, Sox2, Pax6). DISCUSSION:These results indicate that PSCs possess stemness and can be preserved under specific culture conditions. The ability of MSC medium to maintain PSCs in a quiescent and stem-like state provides a valuable model for studying their biology. CONCLUSION:PSCs exhibit stemness, and MSC medium maintains their quiescence, offering a novel experimental platform to study PSC biology and lineage plasticity.
Introduction/ Objective: Spasticity following spinal cord injury (SCI) remains a challenging condition with limited effective treatments. Transplantation of stem cells to replace the lost GABAergic neurons represents a promising therapeutic strategy. Our previous work has confirmed that cerebrospinal fluid-contacting neurons (CSF-cNs) exhibit characteristics of neural stem cells and can differentiate into GABAergic neurons. This study aims to investigate whether transplanted CSF-cNs can differentiate into GABAergic neurons and alleviate spasticity following SCI. METHODS:CSF-cNs were isolated and purified from adult mice in vitro. Following induction culture, differentiation was characterized by immunofluorescence. One week post-SCI, the CSF-cNs were transplanted into the injured area. The survival and differentiation of transplanted cells were assessed via immunofluorescence. Early progression of spasticity was assessed using the hindlimb grasp reflex. Late-stage spasticity relief was evaluated through electrophysiological and gait footprint analysis. RESULTS:Transplanted CSF-cNs survived within the SCI region, with 58% of GFP⁺ cells co-expressing NeuN. These GFP⁺ cells also expressed GABA, the GABA-synthesizing enzyme glutamate decarboxylase (GAD65/67), and the vesicular GABA transporter (VGAT). Within three weeks post-transplantation, the hindlimb grip reflex scores in the transplant group were significantly improved (P < 0.05). By 6 and 12 weeks, the H-reflex rate-dependent depression (RDD) in the transplant group was significantly superior to that of the control group (P < 0.05). Additionally, the stride length, step width, and hindlimb coordination were markedly improved in the transplant group compared to controls. DISCUSSION:This study provides the first experimental evidence that transplanted CSF-cNs survive and differentiate into GABAergic neurons within the SCI microenvironment. Simultaneously, CSFcN transplantation effectively alleviates spasticity following SCI. This outcome aligns with previous findings using stem cell-derived GABAergic progenitor cells for spasticity treatment. Therefore, we hypothesize that the alleviation of spasticity observed in this study may be associated with the differentiation of CSF-cNs into GABAergic neurons. Admittedly, this correlation requires further investigation. Additionally, although both behavioral and electrophysiological results demonstrated significant symptomatic relief, recovery remained incomplete. This indicates that even if CSF-cNs differentiate into GABAergic neurons and supplement the deficient inhibitory neurons in the damaged spinal cord circuits, this process is insufficient to completely reconstruct the complex spinal cord network. Future investigations will explore mechanisms beyond cell replacement, such as the potential role of transplanted cells in modulating synaptic efficacy (for instance, by restoring KCC2 transporter function). Overall, the evidence presented in this study suggests that CSF-cN transplantation represents a viable strategy for alleviating spasticity following SCI. CONCLUSION:Transplanted CSF-cNs can differentiate into GABAergic neurons and that their transplantation can alleviate spasticity in mice following SCI.
Innovations in ear reconstruction in the developing medical field have resulted in various advantages and disadvantages. In addition to rib autologous ear cartilage reconstruction, cartilage differentiation is achieved using stem cell technologies, or cartilage is constructed using tissueengineering-based systems from various natural and artificial materials. Intercalarily, autologous rib ear cartilage reconstruction is routinely used; it is a field that is constantly being developed, from stem cells and cell culture systems to natural and artificial biomaterials and tissue engineering-based systems. It is estimated that 3D scaffold systems created using tissue engineering approaches will continue to be popular for a long time in terms of both shaping and cellularization. The differentiation potential of stem cells, the mechanical strength, biocompatibility, and biodegradability of scaffolds, and especially the use of exosome engineering as a biosignal molecule, which has become popular recently, bring another perspective on cartilage regeneration. The nanostructures of exosomes carry very important information between cells in terms of their anti-inflammatory activities and especially their effects on the proliferation and differentiation activities of stem cells. Especially modified or loaded exosome designs allow different approaches to be created in this field, both for therapeutics and diagnosis. This review offers a regenerative medicine perspective on ear reconstruction studies, addressing the advantages and disadvantages of experimental and clinical designs, including current innovations and artificial organ designs, such as stem cell-exosome strategies. The complexity of the production process, the characterization of biomaterials, the need for standardization of in vitro and in vivo protocols, ethical concerns, and high costs necessitate controlled and comparative analyses of these high-tech products/methods before they are transferred to clinical use.
Tissue Engineering (TE) represents an effective approach for addressing tissue or organ defects and functional impairments. Cells, scaffolds, and growth factors, as the three elements of TE, play an important role in tissue repair. In recent years, there has been growing interest in Wharton's Jelly (WJ), obtained from the umbilical cord of postpartum medical waste. Mesenchymal stem cells, extracellular matrix, and growth factors derived from WJ have been widely used in TE. WJ has shown the potential to treat a wide range of diseases in clinical applications. This review discusses the mechanisms of action of WJ, focusing on their applications in the field of tissue damage repair. We also highlight the advantages, current limitations, and future directions of WJ-based therapies, including challenges related to standardization, scalability, long-term safety, and clinical translation.
INTRODUCTION:Amniotic Fluid Cells (AFCs) have emerged as a promising source for regenerative medicine due to their multipotent properties and accessibility. This study aimed to quantify pluripotency-associated transcript profiles in primary amniocyte cultures and evaluate their associations with gestational age, maternal age, and culture dynamics. METHODS:This study analyzed the expression of ten pluripotency-associated genes (OCT3/OCT4, SOX2, KLF4, MYC, KIT, NANOG, DPPA3, DPPA5, FUT4, and SALL4) in primary AFC cultures derived from 17 pregnancies using quantitative real-time PCR (2-ΔΔCt; ACTB-normalized). Samples were evaluated according to gestational age, maternal age, and fetal sex; culture expandability was assessed based on the maximum passage achieved. AFC cultures were serially subcultured, and RNA was isolated independently at each available passage to quantify longitudinal passage-associated expression changes. Spearman's rank correlation assessed associations between gestational age and maternal age (continuous variables) and expression of SOX2, DPPA5, KIT, NANOG, and SALL4, with Benjamini-Hochberg FDR adjustment (q = 0.05). RESULTS:All investigated markers were detectable across the AFC cultures. In group-based analyses, SOX2, DPPA3, and DPPA5 expression levels were higher in cultures derived from lower gestational ages, while SOX2 and DPPA5 were higher in younger maternal-age groups (p < 0.05). Culture expandability showed positive associations with KIT and selected markers. Serial-passaging analyses suggested that KIT expression was maintained in cultures achieving extended passaging, whereas SOX2 and DPPA5 expression tended to decrease with increasing passage number. DISCUSSION:These findings indicate that developmental timing and in vitro culture dynamics are associated with variability in pluripotency-marker expression in primary amniocyte cultures. Transcript- level profiling may therefore help contextualize inter-individual heterogeneity and passagerelated shifts relevant to stem cell biology and standardized culture selection. CONCLUSION:AFC pluripotency-marker transcripts vary with gestational age, maternal age, and passaging. These transcript-level findings provide a quantitative reference but require protein/functional validation in larger, standardized cohorts.
INTRODUCTION:CAR T therapy is used in various hematological malignancies that involve genetic modification of autologous T cells to target tumor cells. The aim of this meta-analysis was to evaluate the efficacy and safety of CAR T cell therapy in haematological malignancies with relation to response rates, survival outcomes, and treatment-related adverse events. METHODS:This meta-analysis followed PRISMA guidelines. 428 records were identified from Pub- Med, Embase, Web of Science, Scopus, Cochrane Library, and Google Scholar for studies between 2019 and 2024. Clinical trials and observational studies evaluating CAR T therapy in haematological malignancies were included. Study quality was assessed using the Newcastle Ottawa Scale and ROBINS I tool. Random effects models were used, and heterogeneity was measured with the I² statistic. RESULTS:Twelve studies met the inclusion criteria. CAR T therapy showed high ORR and CR in Bcell malignancies, with ORR reaching 90.5% in R/R B NHL and 81% remission in B ALL. CRS was common but mainly mild to moderate, while neurotoxicity and GvHD were less frequent. Metaanalysis demonstrated substantial heterogeneity for ORR, CR, and CRS. Most studies were rated good quality, though moderate bias related to missing data and design differences was observed. DISCUSSION:Meta-analysis shows CAR T therapy achieves high ORR and durable remission in B ALL and DLBCL. CD19-directed therapy was promising. Compared to myeloid cell malignancies, B-cell malignancies showed better response. These findings point towards optimisation of protocol and appropriate patient selection apart from the toxicity management, which is required to improve the outcomes. CONCLUSION:CAR T therapy shows high response rates in haematological malignancies with controllable toxicity, thus establishing its role in relapsed and refractory disease. It highlights the need for improved durability, access, and constant clinical application.
Mesenchymal stem cells (MSCs) are a heterogeneous cell population that can be isolated from various tissues, including adipose tissue and bone marrow. Their ease of isolation and robust ex vivo expansion make MSCs an attractive candidate for regenerative and therapeutic applications, particularly in tissue repair and angiogenesis. Increasing evidence indicates that the therapeutic effects of MSCs are largely mediated through the secretion of extracellular vesicles, especially mesenchymal stem cell-derived exosomes (MSC-Exos). These exosomes contain diverse bioactive molecules, including proteins, nucleic acids, and lipids, which contribute to their regenerative, immunomodulatory, and therapeutic potential. Preclinical studies have demonstrated that MSC-Exos promote tissue repair, regulate immune responses, and alleviate pathological conditions associated with cardiovascular diseases, neurological disorders, musculoskeletal injuries, and immune-mediated diseases. The outcomes of these studies suggest that MSC-Exos represent a promising and effective therapeutic modality for a broad range of diseases. Compared with conventional MSC-based therapies, MSCExos offer several advantages, including reduced immunogenicity, lower tumorigenic risk, improved stability, and easier storage and handling, making them particularly suitable for clinical translation. This review provides a comprehensive overview of the current clinical and translational landscape of MSC-Exos therapy, highlighting their therapeutic applications, underlying mechanisms, and key challenges that must be addressed to facilitate successful clinical implementation. Collectively, the emerging evidence underscores the potential of MSC-Exos to advance regenerative medicine and suggests that exosome-based therapies may represent a transformative approach for the treatment of diverse human diseases.
Mesenchymal stromal cells (MSCs) are multipotent cells of mesodermal origin capable of self-renewal and multilineage differentiation. Characterized by low immunogenicity and tropism toward injury sites, MSCs exhibit critical properties including hematopoietic support, immunomodulation, and tissue regeneration. These unique attributes position MSCs as promising therapeutic tools for hematologic diseases, where disruption of the bone marrow niche impairs normal hematopoiesis. Co-transplantation of MSCs with hematopoietic stem cells (HSCs) facilitates HSC homing to the bone marrow niche and significantly improves post-transplant hematopoietic reconstitution. Furthermore, MSCs show considerable therapeutic potential in both prophylaxis and management of graftversus- host disease (GVHD), a major complication of allogeneic transplantation. The therapeutic mechanisms of MSCs have evolved from an initial focus on engraftment to a broader understanding of their paracrine actions via the "hit-and-run" mechanism, wherein MSCs exert functions through secreted factors and extracellular vesicles before host clearance. Despite these advances, clinical translation faces significant challenges, including poor homing efficiency, cellular heterogeneity, culture-induced senescence, and vulnerability to inflammatory and oxidative stress. This review summarizes clinical applications of MSCs in aplastic anemia, leukemia, and co-transplantation with HSCs, while critically evaluating the balance between therapeutic efficacy and potential risks. Additionally, we discuss emerging bioengineering strategies designed to overcome current limitations and enhance MSC therapeutic potency for next-generation cell therapies in hematologic diseases.
Retinal degenerative diseases like AMD and DR lead to irreversible vision loss through disrupted homeostasis. Mesenchymal Stem Cell-derived Exosomes (MSC-Exo), as a cell-free therapy, demonstrate significant potential in restoring retinal balance via multi-target regulation. This review elucidates the protective effects of MSC-Exo on key retinal cells, including pigment epithelium, endothelial cells, Müller glia, microglia, and ganglion cells, and specifically examines its role in modulating shared cellular pathways such as PI3K/AKT-Nrf2. It highlights a therapeutic cascade from vascular repair and anti-inflammation to neuroprotection, which coordinates intercellular interactions and promotes holistic retinal recovery. Finally, the review discusses the challenges and future clinical prospects of MSC-Exo-based therapies.
INTRODUCTION:Sepsis is a life-threatening condition and ranks among the leading causes of death worldwide. Bone marrow mesenchymal stem cells (BMSCs) have shown promise as a therapeutic strategy for sepsis due to their anti-inflammatory and immune-regulatory properties. However, the precise molecular mechanisms by which BMSCs exert these beneficial effects in sepsis are not yet fully elucidated. METHODS:We developed an in vitro macrophage injury model by co-culturing injured macrophages with BMSCs to evaluate pyroptosis. In vivo, we induced a sepsis model and subsequently transplanted BMSCs. The expressions of proteins involved in the NLRP3 (nod-like receptor family pyrin domain containing-3)-Caspase-1/Caspase-11-Gasdermin D (GSDMD) signaling pathway, as well as pyroptosis in peritoneal macrophages, were then investigated. RESULTS:Our results demonstrated that BMSCs co-cultured with lipopolysaccharide (LPS)-stimulated macrophages improved macrophage viability and reduced pyroptosis in vitro. Moreover, transplantation of BMSCs significantly decreased pyroptosis in peritoneal macrophages and improved the survival rate of sepsis rats. BMSC treatment also downregulated the expression levels of pyroptosis-related proteins in macrophages isolated from sepsis rats, including NLRP3, Caspase-1, Caspase-11, and GSDMD. DISCUSSION:These findings elucidate the mechanisms by which BMSCs exert therapeutic effects in sepsis, particularly through modulation of peritoneal macrophage pyroptosis. This provides a foundation for further research on sepsis and the potential clinical application of BMSCs. A limitation of this study is that the experimental results were derived from a rat model rather than non-human primates. CONCLUSION:This study demonstrates that transplantation of BMSCs may suppress NLRP3-Caspase- 1/Caspase-11-GSDMD-mediated pyroptosis in peritoneal macrophages during sepsis, providing new mechanistic insights into the therapeutic effects of BMSCs.
INTRODUCTION:The hostile inflammatory microenvironment at the injury site often limits the therapeutic potential of human Umbilical Cord Mesenchymal Stem Cells (UC-MSCs) in colitis. To address this challenge, we explored whether priming UC-MSCs with conditioned medium from M1 macrophages (M1-CM) could attenuate detrimental macrophage crosstalk and amplify their immunomodulatory efficacy, improving overall treatment outcomes. METHODS:The M1 macrophage phenotype was induced with LPS and IFN-γ, and M1-CM was collected. UC-MSCs were preconditioned with M1-CM to generate M1UC-MSCs. To assess the therapeutic potential of M1UC-MSCs, a dextran sulfate sodium (DSS)-induced colitis model was established in ICR mice. RESULTS:Preconditioning with M1-CM significantly enhanced the anti-inflammatory properties of UC-MSCs without compromising viability. In vivo, M1UC-MSCs exhibited improved survival and more effectively ameliorated colitis symptoms, reducing disease severity and inflammatory damage compared to non-primed cells. DISCUSSION:These results indicate that inflammatory preconditioning augments the therapeutic function of UC-MSCs by enhancing their adaptability and reducing deleterious interplay with macrophages. This approach is consistent with current strategies to improve cellular tolerance and microenvironmental compatibility. Study limitations include the need for further mechanistic investigation, and subsequent research should prioritize elucidating the specific molecular pathways modulating macrophage-MSC crosstalk. CONCLUSION:M1-CM preconditioning represents a novel and potent strategy to enhance UC-MSCbased therapy for colitis, primarily by improving cellular adaptation and harmonizing the immune response under inflammatory conditions. This method holds significant promise for advancing regenerative medicine applications in colitis.
INTRODUCTION:We tested whether early administration of multiple doses of adipose-derived mesenchymal stem cells (ADMSCs) overexpressing cellular prion protein (PrPC-OVE) could preserve lung parenchyma and function in a rodent model of bleomycin-induced pulmonary fibrosis (PF). METHODS:Cell culture, immunohistochemistry, immunofluorescence, and western blot analyses were conducted. Animals were categorized into sham-control (Group 1), PF (Group 2), PF + ADMSCs (one dose; Group 3), PF + ADMSCs (three doses; Group 4), PF + PrPC-OVE in ADMSCs (one dose; Group 5), and PF + PrPC-OVE in ADMSCs (three doses; Group 6). RESULTS:In vitro, bleomycin suppressed L2 cell proliferation and increased apoptosis and epithelial-mesenchymal transition (EMT) markers, which were significantly reversed by ADMSCs and further enhanced by PrPC-OVE in ADMSCs. TGF-β/Smads signaling-mediated EMT was identified as a crucial mechanism in cellular fibrosis and PF, which was significantly suppressed by silencing TGF-β in L2 cells and PrPC-OVE in ADMSCs co-cultures and lung tissue. By days 28 and 42 after PF induction, O₂ saturation (%) was highest in Group 1, lowest in Group 2, and notably higher in Group 6 than in Groups 3-5. Right-ventricular (RV) systolic blood pressure showed an inverse trend. RV and left lung weights-to-tibial length ratios were significantly greater in Group 2 at both time points. By day 42, Group 2 exhibited the highest lung injury, fibrosis, and protein levels of fibrotic, EMT, inflammatory, and oxidative stress markers, whereas these parameters were lowest in Group 1 and significantly higher in Group 4 than in Group 6. DISCUSSION:This study, which investigated the therapeutic role of PrPC-OVE in ADMSCs in protecting lung parenchyma against bleomycin-induced damage, provides several important preclinical insights. First, the in vitro study demonstrated that bleomycin activated TGF-β/Smads signaling, inducing EMT upregulation in lung epithelial cells (L2 cell line), which plays a crucial role in the initiation and propagation of PF. Second, both in vitro and in vivo studies showed that PrPC-OVE played a fundamental role in attenuating bleomycin-induced PF, mainly through regulation of TGF-β/Smads signaling. Third, ADMSCs were less effective than PrPC-OVE in ADMSCs, and repeated doses of ADMSCs were less effective than repeated doses of PrPC-OVE in ADMSCs in protecting lung function and parenchyma against bleomycin-induced damage. CONCLUSION:Repeated PrPC-OVE-ADMSCs effectively preserved lung function and parenchyma by suppressing TGF-β/Smads signaling in bleomycin-induced PF.
INTRODUCTION:Mesenchymal Stem Cells (MSCs) undergo prolonged in vitro passaging, which leads to cellular senescence, and this reduces their therapeutic effectiveness. This study aimed to investigate whether priming MSCs with a defined cytokine mix under hypoxic environments could reduce senescence and augment their therapeutic potential. METHODS:Human umbilical cord-derived MSCs (UC-MSCs) at passage 4 (P4) and passage 8 (P8) were primed under hypoxic settings with three key cytokines (interferon γ, basic fibroblast growth factor, and leukemia inhibitory factor) for a duration of 24 h. We utilized flow cytometry, polymerase chain reaction, enzyme-linked immunosorbent assay, and other experimental approaches to examine the effects of passaging and priming on cellular properties, immunomodulatory capabilities, and neurotrophic qualities and angiogenic potentials. RESULTS:At passage 8, UC-MSCs showed a loss of their usual spindle-shaped morphology, accompanied by an increase in size, a decrease in proliferative capacity, and a reduction in the expression of stemness-related genes, whereas markers of cellular senescence were enhanced. Nonetheless, priming strategies effectively reduced the expression of these senescence markers, maintained stemness, and augmented the expression of immunomodulatory cytokines, neurotrophic factors, and vasculogenic factors. Both UC-MSCs and primed UC-MSCs at P4 and P8 showed similar surface markers, indicating that neither serial expansion nor priming changed the fundamental immunophenotype of the MSCs. DISCUSSION:The results show that hypoxia-cytokines priming effectively mitigates senescence caused by expansion while keeping their immunophenotype. This strategy not only maintains stem cell properties but also enhances their therapeutic potential, making it a feasible approach to obtain large numbers of highly active MSCs for clinical applications. CONCLUSION:In summary, hypoxia-cytokine priming delayed cellular senescence and possibly enhanced the immunomodulatory, neurotrophic, and vasculogenic properties of UC-MSCs.
INTRODUCTION:Temporomandibular Joint Osteoarthritis (TMJOA) is a degenerative disease mainly characterized by cartilage degeneration. Regeneration of condylar cartilage plays a crucial role in enhancing joint function and mitigating the advancement of TMJOA. Emodin has shown efficacy in promoting osteogenesis and inhibiting extracellular matrix degradation, suggesting its potential as a novel therapeutic approach for promoting condylar cartilage regeneration. METHODS:In this study, we isolated and characterized distinct Fibrocartilage Stem Cells (FCSCs) from the condyles of Sprague Dawley rats. Proliferation and differentiation of FCSCs were assessed using the Cell Counting Kit-8 (CCK-8) assay and two-dimensional (2D) monolayer cultures. Western blot and Quantitative Real-Time PCR (qRT-PCR) analyses were conducted to evaluate the expression levels of bone and cartilage anabolic metabolism markers. RESULTS:The CCK-8 assay demonstrated a dose-dependent increase in cell proliferation, and the peak cell viability was observed at approximately 20 μM (P < 0.01). Analysis using qRT-PCR revealed significant upregulation of the fibrocartilage-specific marker, specifically Col I (P < 0.0001), alongside the concurrent downregulation of adipogenic markers Ppar-γ and LPL (P < 0.05), as well as osteogenic markers ACAN and ALP. Western blot analysis further confirmed elevated levels of Col I and reduced levels of ACAN and ALP following treatment with 20 μM. DISCUSSION:Our findings demonstrated that emodin selectively promoted the differentiation of FCSCs towards chondrogenic lineages within a specific concentration range in vitro. CONCLUSION:In conclusion, emodin enhanced the fibrocartilage-specific differentiation of FCSCs, indicating the potential therapeutic application of emodin in regenerating cartilage in the Temporomandibular Joint (TMJ).
