Osteoarthritis (OA), a prevalent degenerative joint disorder, is suboptimally managed by existing treatments owing to adverse effects, limited therapeutic efficacy, and the inability to achieve genuine cartilage repair. Adipose-derived stromal vascular fraction (SVF)-gel, which is abundant in stem cells and bioactive factors, has an autologous origin and favorable immunocompatibility and exhibits favorable safety and efficacy in facilitating cartilage regeneration and repair. Building upon prior research, this review discusses the clinical efficacy and potential of SVF-gel, while also recognizing challenges such as the standardization of preparation procedures and dosage optimization. Future research ought to concentrate on integrating advanced technologies, including gene editing, artificial intelligence (AI), and nanomaterials, to drive its development towards more precise and personalized therapeutic approaches.
Background:Building on our previous work, we found that human omental adipose-derived mesenchymal stem cells (ADSCs) promote ovarian cancer growth and metastasis by inducing PAX8, a key oncogenic transcription factor in ovarian cancer. TAZ is a core downstream effector of the Hippo signaling pathway, closely associated with malignant progression in multiple tumors, and our prior work confirmed that PAX8 can stabilize TAZ protein. In this study, we aimed to further clarify how ADSCs regulate PAX8 to explore the downstream signaling pathway driving malignant progression in ovarian cancer. Materials and Methods:Ovarian cancer cells were treated with ADSC-conditioned medium (ADSC-CM) to investigate the regulatory effect of ADSCs on miR-214-5p expression. qRT-PCR and western blot were applied to detect the levels of miR-214-5p, PAX8, β-catenin, and TAZ, so as to dissect the regulatory relationship of ADSCs/miR-214-5p/PAX8/β-catenin/TAZ. A dual-luciferase reporter assay verified the direct targeting interaction between miR-214-5p and PAX8. CCK-8 and Transwell invasion assays were conducted to assess the biological functions of the miR-214-5p/PAX8 axis in ovarian cancer progression. Results:We show that ADSC treatment significantly downregulated miR-214-5p in ovarian cancer cells. Moreover, PAX8 is a direct target of miR-214-5p: knockdown or overexpression of miR-214-5p altered PAX8 expression by directly binding to PAX8 mRNA. Additionally, PAX8 stabilizes TAZ by activating the β-catenin pathway. Functionally, miR-214-5p inhibited ovarian cancer cell proliferation and invasion by downregulating PAX8. Conclusion:Taken together, ADSCs downregulate miR-214-5p to increase PAX8 expression, which sequentially activates the β-catenin pathway and stabilizes TAZ. This novel signaling axis is responsible for ADSC-induced ovarian cancer growth and metastasis.
Background:Preclinical evidence supports the potential use of human umbilical cord blood-derived mononuclear cells (hUCB-MNCs) for preterm infants with preterm birth-associated complications (PBAC). Allogeneic hUCB-MNCs could extend this therapeutic option to infants without autologous cord blood available at birth. Methods:This phase I, open-label, single-arm, and single-center study enrolled 10 preterm infants (gestational age [GA] 27.1 ± 0.7 weeks) with two or more PBAC. All received a single intravenous infusion of allogeneic, partially or fully human leukocyte antigen (HLA)-mismatched hUCB-MNCs without preconditioning or immunosuppression. The median cell was (3.51 ± 0.93) × 106 cells/kg. Safety, adverse events, and clinical outcomes were assessed over a 2-year follow-up period. Results:All infants survived intensive care unit discharge. Four infants were lost to follow-up for parental reasons, with no in-hospital adverse events noted, their long-term outcomes remain unconfirmed. No infusion-related adverse events, acute reactions, or graft-versus-host disease (GVHD) were detected. One case of cerebral palsy was diagnosed during follow-up and was unrelated to cell infusion. Five of the six infants with evaluable long-term data had no respiratory, developmental, or neurological complications. Plasma inflammatory cytokines exhibited exploratory changes after infusion, without confirmed causality. All infants with follow-up data reached age-appropriate growth at 12 and 24 months of corrected age. Conclusions:No major infusion-related adverse events were observed in preterm infants with PBAC receiving allogeneic hUCB-MNC infusion. Six infants completed 2-year follow-up and four were lost to follow-up. Constrained by small sample size and attrition, the results need further verification in larger controlled trials. Trial Registration:Chinese Clinical Trial Registry: ChiCTR-OPN-15006932, ChiCTR2000035227.
Background:Mesenchymal stem cells (MSCs) are multipotent, nonhematopoietic progenitors capable of supporting hematopoiesis and regulating immunity. Clinical studies have shown that MSCs can aid hematopoietic recovery and balance T helper 17 (Th17) and Treg cells in some aplastic anemia (AA) patients refractory to immunosuppressive therapy (IST). However, full clinical efficacy remains unachieved. To enhance MSC immune regulation while ensuring safety, previous research found interleukin-10 (IL-10), an immunosuppressive factor, can boost MSC function. In this study, we used lentiviral transduction to induce IL-10 gene overexpression in human umbilical cord-derived MSCs and validated their efficacy and safety in AA mouse models. Methods:MSCs were isolated from umbilical cords and genetically modified via lentiviral transduction to overexpress IL-10, generating OEIL10-MSCs. Empty vector plasmids (pCDH-CMV-MCS-EF1-copGFP) were transduced into MSC to create PCDH-MSCs. Both in vitro and in vivo experiments were conducted to investigate their functional changes and mechanisms. Mouse models of nonsevere AA (NSAA) and severe AA (SAA) were established and treated with OEIL10-MSCs or control PCDH-MSCs. The therapeutic efficacy of OEIL10-MSCs in AA was evaluated by assessing blood cell counts, Th17/Treg ratios, and bone marrow histopathology in an AA mouse model. Safety was assessed through liver and kidney function tests. Results:In vitro experiments verified that OEIL10-MSCs exhibited enhanced proliferation capacity, improved anti-inflammatory ability, and reduced oxidative stress levels. In NSAA mouse models, OEIL10-MSCs were more effective in restoring hematopoietic function and elevating Treg cell proportions. In contrast, in SAA mouse models, OEIL10-MSCs demonstrated superior efficacy in elevating red blood cell (RBC) levels and reducing DNA damage compared to PCDH-MSCs. Conclusion:Overexpression of IL-10 may enhance the efficacy of MSCs in treating AA, presenting a promising therapeutic strategy for the clinical application of MSCs.
Mesenchymal stem cells (MSCs) and regulatory T cells (Tregs) form a key immunoregulatory axis essential for maintaining immune homeostasis and treating autoimmune diseases, transplant rejection, and inflammatory disorders. Although both MSCs and Tregs have been individually studied, a clear synthesis of how MSCs regulate Treg function across distinct mechanistic layers and disease contexts remains lacking. MSCs regulate Treg differentiation, expansion, and functional stability through paracrine signaling, intercellular contact-dependent pathways, extracellular vesicle (EV)-mediated transfer of microRNAs (miRNAs) and proteins, mitochondrial transfer, and metabolic and epigenetic reprogramming. These mechanisms restore Th17/Treg balance, suppress inflammation, and promote tissue repair. Preclinical studies demonstrate strong therapeutic potential in multiple immune-mediated diseases; however, clinical translation remains limited. Unlike previous studies, this review integrates soluble and exosomal signaling pathways, compares shared and disease-specific regulatory mechanisms, and critically evaluates MSC source variability, methodological limitations, and causes of clinical inconsistency. It aims to provide a conceptual framework to guide future mechanistic studies and clinical development of MSC-Treg-based therapies.
Substance use disorders (SUDs) impose major global morbidity and mortality, yet the cellular mechanisms linking genetic risk to neural vulnerability, disrupted neurodevelopment, and drug-induced neuroadaptations remain poorly understood. Human pluripotent stem cell (hPSC) technologies, including embryonic and induced pluripotent stem cell (ESC/iPSC)-derived neurons, three-dimensional (3D) brain organoids, and organoid-on-a-chip platforms, provide scalable, human-relevant models to address this gap. ESC/iPSC-derived neuronal cultures enable interrogation of genetic and epigenetic determinants of drug susceptibility and response; cerebral organoids recapitulate tissue architecture and emergent network dynamics; and microfluidic organoid-on-a-chip systems facilitate maturation, enhance reproducibility, and enable controlled exposure paradigms. In this review, we synthesize recent stem cell-based studies of alcohol, opioid, and stimulant exposure, highlighting insights into neurodevelopmental disruption, synaptic and signaling alterations, neuroinflammation, and network-level dysfunction. We critically evaluate limitations of stem cell-based studies, including ethical concerns, inter- and intra-line variability, incomplete cellular maturation, and difficulties in modeling complex circuitry and comorbid conditions. We propose strategies to enhance translational relevance, including standardized differentiation protocols, addition of patient-derived cells and vascular and immune components, and integration of multi-omics approaches (transcriptomics, epigenomics, and proteomics) with functional readouts to map molecular pathways underlying drug vulnerability and resilience. Finally, we outline the therapeutic and precision-medicine potential of stem cell platforms for target discovery, predictive toxicology, and individualized treatment modeling. Despite remaining challenges, stem cell-based approaches offer a powerful and increasingly tractable path from genetic association to mechanistic insight and therapeutic innovation in addiction research.
Background:This systematic review and meta-analysis aimed to evaluate the effectiveness of stem cell therapies for patients with amyotrophic lateral sclerosis (ALS) based on optimal dosing and administration routes, as well as the safety profiles of stem cells and their derived products. Methods:The review followed PRISMA guidelines and involved a comprehensive literature search up to October 2025, receiving ethical approval from Tabriz University of Medical Sciences and registration in PROSPERO. It utilized international databases, including PubMed/MEDLINE, Embase, Cochrane Library, Scopus, Web of Science, ProQuest, ClinicalTrials.gov, and Science Direct. The included studies comprised randomized controlled trials (RCTs), quasi-experimental studies, and other interventional designs involving ALS patients treated with stem cell therapies. In total, 31 studies were analyzed, featuring 7 controlled trials with 370 participants and 24 non-controlled pre-post studies with 460 participants. Heterogeneity was evaluated using I 2 statistics, and subgroup analyses were conducted based on treatment duration and dosing. Results:A pooled analysis (treatment group: n = 93; control group: n = 90) demonstrated a significant attenuation in the progression of disease severity, as measured by the ALS Functional Rating Scale (ALSFRS), in stem cell groups versus controls (weighted mean difference [WMD]: 8.89 95% CI: 4.12-13.67; p = 0.0003), which was beneficial for both the ≥10 × 106 and <10 × 106 dose sub-groups. However, a meta-analysis of single-arm studies in two control (pre-intervention) and intervention phases (n = 88) demonstrated no significant difference in progression of ALSFRS between study phases by time: month 3 (WMD: -1.27 (-3.01 to 0.47); p = 0.15), month 6 (WMD: -2.69 (-5.62 to 0.25); p = 0.07), month 9 (WMD: -1.55 (-3.49 to 0.39); p = 0.12), and month 12 (WMD: -7.59 (-13.95 to -1.26); p = 0.02). An accelerated decline in forced vital capacity (FVC) was observed during the intervention phase, with statistically significant reductions at month 3 (WMD: -10.91; 95% CI: -16.39 to -5.43; p < 0.0001) and month 6 (WMD: -15.97; 95% CI: -28.60 to -3.33; p = 0.01) compared with the pre-intervention control phase. Nevertheless, sensitivity analyses excluding studies involving high-dose mesenchymal stem cell (MSC) therapies demonstrated that these differences were no longer statistically significant. Moreover, no significant change in progression rate was observed at month 9 (WMD: -8.10 (-18.25 to 2.06); p = 0.12). The route of MSCs administration (intrathecal [IT], intramuscular [IM], and intravenous [IV]) had no effect on the results of ALSFRS and FVC, reinforced by sensitivity analyses. Adverse events were mostly mild, with headaches most frequent in high-dose groups. Conclusion:Stem cell therapy for ALS appears to be safe, with preliminary evidence suggesting potential therapeutic benefit in slowing disease progression in selected patients. Nevertheless, the existing evidence base remains exploratory, and definitive conclusions regarding clinical effectiveness cannot yet be drawn. Future research should prioritize large-scale and multicenter RCTs with standardized cell manufacturing protocols and longer follow-up periods.
Background:Colon cancer (CC) remains a leading cause of cancer-related mortality worldwide, driven largely by the complex interactions within the tumor microenvironment (TME). Fibroblast activation protein (FAP) is highly expressed in cancer-associated fibroblasts (CAFs) and is associated with poor prognosis, yet its role in coordinating immune evasion and cancer stemness remains to be fully elucidated. Methods:We integrated multiomics data from TCGA and GEO databases, utilizing bulk RNA-seq and single-cell RNA-seq (scRNA-seq) analyses. Findings were validated via tissue microarray (TMA) immunohistochemistry (IHC). We further employed cell-cell communication analysis, pseudotime trajectory modeling, and the Connectivity Map (CMap) for drug sensitivity prediction and molecular docking. Results:FAP was significantly upregulated in CC tissues and correlated with advanced clinical stages. scRNA-seq confirmed that FAP is predominantly expressed in CAFs. Functional analysis revealed that FAP-high tumors are enriched in extracellular matrix (ECM) remodeling and immunosuppressive pathways. Cell-cell communication analysis identified that FAP+ CAFs interact with T cells and cancer stem cells (CSCs) primarily through the COL1A1/2-CD44 axis. Specifically, FAP+ CAFs promote the differentiation of naive T cells into regulatory T cells (Tregs) and are positively correlated with various stemness markers, including CD44, ABCG2, and BMI1. Based on these findings, we established a 14-gene prognostic risk model with robust predictive accuracy (area under the curve [AUC] > 0.64) and identified AS604850 and LY364947 as potential therapeutic agents. Conclusion:Our study demonstrates that FAP+ CAFs orchestrate a dual-functional "immunosuppressive stem cell niche" via the COL1A1/2-CD44 signaling axis. Targeting this FAP-driven niche provides a promising strategy for overcoming immunotherapy resistance and improving clinical outcomes in CC.
Background:Hepatocellular carcinoma (HCC) suffers from a poor prognosis largely due to its profound molecular heterogeneity and high frequency of relapse, challenges that are closely linked to the biology of cancer stem cells (CSCs) and a lack of effective stemness-related prognostic biomarkers. Identifying CSC-related prognostic biomarkers and therapeutic targets is critical for improving patient outcomes. Methods:We integrated differential expression analysis, weighted gene co-expression network analysis (WGCNA), and CSC gene databases to identify core prognostic genes driven by stemness mechanisms. A robust prognostic model was developed and validated using multiple machine learning algorithms across TCGA and Gene Expression Omnibus (GEO) cohorts. The clinical relevance of the signature was assessed via receiver operating characteristic curve (ROC) curves, survival analysis, and association with tumor stage. Single-cell RNA sequencing (scRNA-seq) and computational drug repositioning coupled with molecular docking were employed to explore mechanistic insights and therapeutic candidates. Results:Intersection analysis identified 12 core genes enriched in CSC-associated pathways. The optimal CoxBoost model demonstrated superior predictive performance for overall survival (OS) in internal, external, and meta-analyses. The signature's single-sample GSEA (ssGSEA) score exhibited high diagnostic accuracy, correlated with advanced tumor stage, and enabled effective risk stratification. Single-cell analysis revealed DARS2 enrichment in M1 macrophages, suggesting a role for CSCs in modulating the tumor immune microenvironment. The histone deacetylase (HDAC) inhibitor belinostat was prioritized via Drug Signature Database (DSigDB) screening and validated by molecular docking as a candidate for targeting the CSC-related signature. Conclusion:This study establishes a novel CSC-associated gene signature for diagnosis and prognosis in HCC and nominates belinostat as a repurposing candidate for targeting stemness-related pathways, offering a promising strategy for personalized therapy.
Background:Endometriosis is a chronic, estrogen-dependent inflammatory disorder characterized by the ectopic implantation of endometrial-like tissue. Although retrograde menstruation is highly prevalent, only a subset of women develops the disease. This epidemiologic paradox suggests that intrinsic molecular alterations in the eutopic endometrium may precondition refluxed cells to survive under inflammatory and oxidative stress. Methods:Eutopic endometrial transcriptomes from the GSE6364 dataset (21 endometriosis patients and 16 controls) were analyzed across menstrual phases. Differential expression analysis, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment, and hallmark inflammatory gene set profiling were performed. Protein-protein interaction (PPI) networks and three machine-learning algorithms maximal clique centrality (MCC), random forest (RF), and least absolute shrinkage and selection operator (LASSO) were applied to identify robust feature genes. Immune cell infiltration was estimated using CIBERSORT . Excision repair cross-complementation group 1 (ERCC1) was further evaluated via cross-species evolutionary conservation and in silico structural modeling of high-risk variants. Additionally, an exploratory stemness-related single-sample gene set enrichment analysis (ssGSEA) was conducted, and correlations between the stemness score and the feature genes were examined. Results:A total of 443 differentially expressed genes (DEGs) were identified, which were significantly enriched in inflammatory cascades, including the NF-κB, Toll-like receptor, and cytokine signaling pathways. Four convergent feature genes ERCC1, SOX3, P75NTR (encoded by NGFR), and FPR1 were prioritized. Immune deconvolution revealed selective immune remodeling in the eutopic endometrium, characterized by elevated activated natural killer (NK) cells and reduced CD8+ T cells, which correlated significantly with the expression of the feature genes. ERCC1 exhibited high evolutionary conservation, and structural modeling of missense variants predicted the disruption of protein function within conserved DNA repair domains. Exploratory stemness analysis revealed no significant overall case-control difference after adjusting for menstrual phase, though the early secretory subset exhibited a marginally higher score in endometriosis. Conclusions:These findings highlight a stress-adaptive transcriptomic state in the eutopic endometrium driven by inflammatory signaling, selective immune remodeling, and altered DNA repair capacity. Specifically, ERCC1 may serve as a critical mechanistic link between inflammatory pressure and impaired genomic maintenance, thereby facilitating cellular persistence and lesion establishment. Furthermore, our data indicate that this four-gene signature primarily reflects a DNA-repair-adaptive program rather than a global bulk-tissue stemness shift.
Human papillomavirus (HPV) infection is the most common sexually transmitted viral infection, strongly associated with chronic inflammation and cervical cancer progression in women. Persistent HPV infection leads to an inflammatory microenvironment that promotes epithelial dysplasia and immune evasion. Exosomes derived from mesenchymal stem cells (MSC-exosomes) have emerged as promising immunomodulatory and anti-inflammatory agents. This review examines current evidence on the interaction between HPV-induced inflammation and exosomal signaling, with a particular focus on the therapeutic potential of MSC-exosomes. We discuss their roles in immune regulation, miRNA delivery, suppression of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling, and epithelial regeneration. This anti-inflammatory effect may also impair local immune surveillance, potentially enabling viral persistence and progression to malignancy. In this narrative review, We reviewed PubMed, Scopus, and Web of Science articles published up to 2024 on MSC-exosome interactions with immune regulation in HPV-related diseases. Evidence suggests that while the anti-inflammatory effects of MSC-exosomes may help control HPV-associated inflammation, they can also impair local immune surveillance, potentially facilitating viral persistence and progression toward malignancy. This dual activity positions MSC-exosomes as a double-edged sword in the context of HPV pathogenesis. A deeper understanding of this paradox is essential for designing safer, context-specific MSC-based therapies that balance anti-inflammatory benefits with effective antiviral immune responses. Although direct studies on MSC-exosomes in HPV infections are limited, existing models suggest that they have the capacity to attenuate inflammation and restore cervical tissue homeostasis. This article also highlights knowledge gaps and future research directions necessary to develop MSC-exosome-based therapies for HPV-related cervical diseases.
Introduction:Articular cartilage degeneration can lead to a progressive loss of joint function and, eventually, total joint failure. The use of mesenchymal stem cells (MSCs) for cartilage regeneration has proven effective; however, the difficulty of their clinical application has prompted research into other strategies, such as using MSC secretome instead of cells themselves. Our aim was to generate a culture system to obtain large amounts of MSC secretome with a controlled and known composition and the potential to induce articular cartilage regeneration. Methods:Human dental pulp stem cells (hDPSCs) embedded in alginate/agarose beads were cultured in a bioreactor with constant agitation and periodic culture medium exchange. The secretome was collected and characterized according to known chondral markers (TGF-β1, SERPINE1, miR-140, miR-675, miR-23a, miR-204, miR-211, and miR-337-5p). Metabolomic and proteomic analyses were carried out for detailed characterization of the collected secretome. The chondrogenic potential of the collected secretome was tested in an in vitro model developed using human primary chondrocytes cultured in 3D on alginate-agarose scaffolds. Results:Our system allowed us to obtain 1200 mL of secretome from intensive cultures of hDPSCs. The cells cultured on the platform remained viable and in an active anabolic state, secreting large amounts of prochondrogenic mediators. Variability between batches of secretome was low, and the collected secretome induced primary chondrocyte differentiation in vitro. Conclusions:Genomic, metabolomic, and proteomic data indicate that hDPSCs on the platform proliferate and acquire a chondrocyte-like phenotype. These cells secrete mediators that define a microenvironment favorable for chondral regeneration. This is further supported by evidence found in the in vitro differentiation model. Our platform allows the production of large volumes of secretome with controlled composition and chondrogenic induction potential. This is a necessary preclinical study for the subsequent analysis of the secretome obtained using in vivo experimental models.
Diabetic nephropathy (DN) remains a major cause of end-stage kidney disease. Stem cell-derived exosomes have emerged as a promising therapeutic strategy due to their ability to deliver bioactive molecules to damaged tissues. This narrative review, conducted in accordance with PRISMA guidelines, aims to evaluate the therapeutic potential of exosomes derived from various stem cell sources, including mesenchymal stem cells (MSCs), embryonic stem cells (ESCs), and induced pluripotent stem cells (iPSCs), in the context of DN. A comprehensive literature search was performed using relevant databases (e.g., PubMed, Web of Science, Scopus) to identify preclinical and clinical studies investigating the effects of stem cell-derived exosomes on DN. The identified studies were assessed for quality and methodological rigor. Priority was given to high-impact studies and those with robust experimental evidence. The selected literature was synthesized thematically to provide a coherent overview of the current state of research on exosome-based therapies for DN, highlighting current findings, and future directions. Results from preclinical studies suggest that exosomes derived from different stem cell sources can exert reno-protective effects, including reducing inflammation, fibrosis, and oxidative stress. However, comparisons between different exosome types indicate that MSCs-derived exosomes (MSC-Exos) may offer superior therapeutic benefits. While clinical trials are ongoing to evaluate the safety and efficacy of stem cell-derived exosomes in DN patients, further research is needed to optimize exosome production, delivery, and therapeutic efficacy.
PurposeInfluence of bone marrow mesenchymal stem cells (BMSCs)-derived exosomes (Exo) on fracture healing in diabetes mellitus (DM) was investigated.MethodsSerum LSD1, SIRT3, superoxide dismutase (SOD), and malondialdehyde (MDA) were detected in patients with diabetic fractures. Under high-glucose (HG) conditions, influences of LSD1, SIRT3, BMSCs-derived Exo, and BMSCs-derived Exo harboring miR-329-3p on osteogenic differentiation and oxidative stress of MC3T3-E1 cells were assayed via ALP staining, western blotting, dichlorofluorescin diacetate (DCFH-DA) staining, and enzyme-linked immunosorbent assay (ELISA). Mice with diabetic fractures were treated by BMSCs, BMSCs-derived Exo, or a combination of BMSCs and GW4869. Western blotting and immunohistochemistry examined protein expression in fracture tissues.ResultsDM patients with nonunion showed higher serum LSD1 and MDA and lower serum SIRT3 and SOD than those with normal fracture healing. In HG-induced MC3T3-E1 cells, LSD1 silencing upregulated SIRT3, RUNX2, OPG, and SOD, increased mature osteoblasts, and reduced reactive oxygen species (ROS) and MDA; SIRT3 silencing revered these results. LSD1 downregulated SIRT3, RUNX2, OPG, and SOD, reduced mature osteoblasts, and enhanced ROS and MDA in HG-induced MC3T3-E1 cells; BMSCs-derived Exo abrogated these influences. BMSCs-derived Exo harboring miR-329-3p suppressed LSD1 and ROS, upregulated SIRT3, RUNX2, and OPG, and increased mature osteoblasts in HG-induced MC3T3-E1 cells. BMSCs transplantation in diabetic fracture mice elevated SIRT3, ALP, RUNX2, OPG, and miR-329-3p and reduced LSD1 in fracture tissues; miR-329-3p silencing in BMSCs or GW4869 treatment of mice counteracted these effects.ConclusionBMSCs-derived Exo harboring miR-329-3p induces bone regeneration in diabetic fractures by relieving the SIRT3-mediated oxidative stress via inhibiting LSD1. It has potential in treating diabetic fractures.
In the fat grafting procedure, the new European Regulations and the necessity to perform minimal manipulation processing require the verification of security and performance levels of each device and, in this particular context, functional and tissue responses. The aim of this study is to evaluate the integrity of adipocytes and adipose-derived stromal/stem cells (ADSCs) after their processing using two new mechanical devices operating with a closed system. In this study, two new fat processing systems were assessed. We also considered the biological results of the processing and compared them with the results obtained from Coleman’s procedure, the golden standard procedure in fat processing. Histological evaluations revealed the preservation of the fat morphology after both processing types, with similar cellular yields of the extracted ADSCs. The adopted techniques enable the isolation of ADSCs with robust differentiation potential toward adipogenic, chondrogenic, and osteogenic lineages. Furthermore, adipose tissue samples exhibit high efficiency in extracellular vesicle secretion, indicating a promising potential for therapeutic and regenerative applications. The results suggest that the new systems allow the preservation of fat morphology, stemness, and regenerative potential. The novel processing technique proposed by the authors consists of a closed-loop mechanical system that combines continuous filtration and emulsification of adipose tissue through saline washing, aimed at removing biological debris and residual pharmacological agents.
Background:Soft tissue defects refer to defects in or damage to human soft tissue, which may affect the appearance, function, and health of the human body and cause physical and psychological distress to the patient. Hyaluronic acid (HA), a fillable material in the field of plastic medicine, could play a crucial role in reconstructing soft tissues and enhancing the adipogenic differentiation of adipose-derived stem cells (ADSCs). Methods:We isolated ADSCs from human subcutaneous adipose tissue and cultured them in HA hydrogel at a density of 2.1 × 106/mL, followed by subcutaneous injections of ADSCs (0.2 mL, 1 × 107 cells/mL with 0.2 mL of DMEM) and HA hydrogel (0.2 mL) into the dorsal side of nude mice. The expression of related genes and proteins was detected using RT‒qPCR and western blotting. The adipogenic differentiation of ADSCs and adipogenesis in nude mice were assessed using CCK-8, Oil Red O, and HE staining. Results:The results revealed an increase in the levels of miR-181a-5p and adipocyte differentiation-related proteins (PPARγ, C/EBPα, FABP4, and adiponectin) and a decrease in the levels of autophagy-related proteins (LC3II/I, Beclin-1, and ATG5) in ADSCs cocultured with HA hydrogel. The adipogenic differentiation ability of ADSCs was enhanced. From a mechanistic standpoint, HA hydrogel inhibited the expression of ATG5 by promoting the expression of miR-181a-5p, thus inhibiting autophagy, promoting the adipogenic differentiation of ADSCs, and promoting the formation of adipose tissue in vivo. Conclusions:Our findings suggest that a combined culture of ADSCs and HA hydrogel may be a new method to enhance the adipogenic differentiation of ADSCs.
Somatic cell nuclear transfer (SCNT) holds great promise for regenerative medicine and agriculture, but its application is severely hampered by low efficiency, primarily attributable to aberrant epigenetic reprogramming. Although embryonic stem cells (ESCs) and trophoblast stem cells (TSCs) have been successfully derived from cloned embryos, an in vitro counterpart of the primitive endoderm (PrE) lineage has remained unavailable. To address this gap, this study reports the first successful establishment of extra-embryonic endoderm stem cell lines (XENs) from mouse SCNT-derived blastocysts (NT-XENs). Under conventional culture conditions, NT-XENs were generated from hybrid B6D2F1 blastocysts at a high efficiency of 55%, statistically comparable to that of fertilization-derived XEN lines (FD-XENs, 50%), whereas derivation from inbred C57BL/6J SCNT-derived blastocysts was markedly lower (12.5%). Immunofluorescence and NanoString multiplex gene expression profiling confirmed that NT-XENs robustly expressed specific marker genes for PrE/XENs (e.g., Gata4, Gata6, and Sox17), while exhibiting negligible or absent expression of pluripotency and trophoblast markers. Based on NanoString assay data, NT-XENs and FD-XENs shared highly similar gene expression patterns, yet also exhibited some nonnegligible differences, exemplified by the differentially expressed genes (DEGs) Pecam1, Gtl2, Thbd, and Xlr3b. These differences raise a preliminary hypothesis that the NT-XENs might exhibit a slight transcriptional propensity toward a more differentiated state, and potentially reflect lingering traces of SCNT-associated epigenetic errors, such as localized dysregulation of imprinted genes and X-linked transcripts. In summary, this study successfully establishes NT-XEN cell lines, providing a valuable in vitro model for investigating the reprogramming scenarios of PrE lineage in SCNT and the mechanisms underlying developmental failure of cloned embryos.
This comprehensive review examines the interconnected roles of apoptosis, neurogenesis, and stem cell therapy in neuroprotection and neurorestoration following intracerebral hemorrhage (ICH). We emphasize that these processes should be interpreted within a unified secondary injury cascade rather than as independent topics. Following ICH, hematoma toxicity, erythrocyte lysis, iron overload, oxidative stress, inflammation, blood-brain barrier disruption, and perihematomal edema interact to activate multiple regulated cell death pathways. Apoptosis remains important, but ferroptosis and other inflammatory cell-death programs are also increasingly relevant to ICH research. The endogenous neurogenic response following ICH is analyzed, highlighting both its potential and its marked limitations in spontaneous recovery, particularly the poor survival and functional integration of newborn cells. We then examine how stem cell therapy may bridge antiapoptotic neuroprotection and proneurogenic neurorestoration through paracrine signaling, microenvironment modulation, and limited cell replacement. Current diagnostic advances and clinical trial designs for stem cell therapy in ICH are reviewed, but we also discuss major translational barriers, including weak distinction between rodent and human evidence, limited cell survival, delivery-route constraints, immune compatibility, source heterogeneity, and tumorigenicity. Finally, we propose that future progress will require stage-specific and clinically translatable strategies rather than isolated pathway targeting. This integrated framework may help refine therapeutic priorities for ICH patients.
Retinoblastoma (RB), a malignant intraocular tumor, represents a critical public health challenge, particularly for pediatric populations, underscoring the urgent need for innovative therapeutic approaches and novel drug development. In this study, amentoflavone (AMF) was identified as a potential agent against RB cancer stem cells (RBCSCs). While CCK8 assays revealed moderate anti-proliferative effects of AMF on RB cell lines, migration and invasion assays demonstrated its potent ability to suppress cancer cell motility. Tumorsphere formation assays further indicated that AMF significantly reduces RB cell stemness. qRT-PCR analysis showed that AMF downregulates expression of stem cell markers (CD44, CD133, Oct4, and Nanog) in a dose-dependent manner. In vivo studies confirmed that AMF inhibits tumor metastasis and prolongs survival in RB mouse models. To elucidate its mechanism of action, RNA sequencing, molecular docking, and surface plasmon resonance (SPR) were employed. These analyses revealed that AMF directly targets smoothened (SMO) to disrupt the SHh signaling pathway, thereby suppressing RB stem cell (RBSC) self-renewal and tumor progression. This work uncovers a previously unreported mechanism by which AMF hinders RB development and highlights its potential as a promising therapeutic candidate for this aggressive pediatric cancer.
