Therapeutic angiogenesis (TA) is a promising strategy for treating ischemic diseases, mainly by targeting the angiogenesis pathways and cells, particularly VEGF and endothelial progenitor cells. Although stem cell therapy has been extensively investigated, its clinical translation remains limited by challenges such as poor cell retention, low survival rates, and inefficient integration. In this review, we propose a mechanism-based framework of angiogenesis to discuss how biomaterials act synergistically with stem cells mainly through two distinct pathways: enhancing paracrine capacity and promoting direct differentiation of vascular lineage cells for vascular repair. Firstly, we go through the scientific literature and clinical studies, and summary the researches on biomaterials serve as artificial microenvironments to improve the retention and secretory function of mesenchymal stem cells (MSCs) and adipose-derived stem cells (ADSCs), thereby maximizing the release of angiogenic factors such as VEGF, bFGF, NGF, microRNA and so on. Secondly, we explore how functionalized biomaterials guide the in situ recruitment of endothelial progenitor cells (EPCs) and support the structural maturation of induced pluripotent stem cell (iPSC)-derived endothelial cells. By integrating these mechanism-driven approaches, we offer new perspectives on future directions for preclinical research and clinical translation of biomaterial-based therapies. Overall, this review has examined the role of individual stem cells and biomaterials, especially enhanced angiogenesis by stem cells focusing on their mechanisms of action and preclinical and clinical applications. We further discussed the challenges encountered by stem cell therapy in advancing to the stage of clinical transformation and considered future prospects.
Umbilical cord blood (UCB) and umbilical cord tissue (UCT) are non-invasive, readily available sources of stem cells with significant potential for regenerative medicine and hematopoietic transplantation. While hematopoietic stem cells from UCB and mesenchymal stem cells from both UCB and UCT are clinically applied, other cord-derived populations remain under investigation, offering novel therapeutic opportunities alongside translational challenges. This review synthesizes current knowledge on stem cell populations derived from UCB and UCT. Hematopoietic and mesenchymal stem cells have established clinical roles, whereas unrestricted somatic stem cells, embryonic-like stem cells, MUSE cells, and multipotent progenitor cells show preclinical promise. These populations differ in differentiation potential, therapeutic application, and biological characteristics. Translational barriers include limited cell numbers, variable engraftment, immune compatibility, and challenges in long-term preservation. Emerging strategies, such as ex vivo expansion, co-transplantation, and nanoparticle-assisted delivery, aim to enhance efficacy, precision, and safety. This narrative review highlights both opportunities and challenges of umbilical cord stem cell therapies. Standardized protocols, interdisciplinary collaboration, and continued innovation are essential to optimize clinical outcomes and fully realize the translational potential of these diverse populations.
Stem cell therapies hold promise for halting or reversing kidney disease and improving kidney transplant (KTx) outcomes. One route to large-scale clinical application of stem cell therapies for kidney disease is through their capacity to modulate the balance between tissue injury and repair via crosstalk with other cells. Among the key disease-modulating effects of stem cells is their interaction with components of the immune system involved in harmful inflammation during acute kidney injury (AKI), chronic kidney disease (CKD) and complications of KTx. Extensive basic research demonstrates that stem cells employ diverse paracrine mechanisms to re-program immunological activities from pro-inflammatory/pro-fibrotic to anti-inflammatory/pro-repair. The therapeutic benefits of these effects are confirmed in many pre-clinical models of AKI, CKD and KTx for autologous and allogeneic stem cells including hematopoietic stem cells, mesenchymal stem cells, renal progenitor cells, and induced pluripotent stem cells. Nonetheless, translating these findings into therapeutic immunomodulatory cell products that improve the lives of those with kidney disease is highly challenging. The aims of this review are to: (a) Summarize recent insights into the common molecular and cellular mechanisms of immune-mediated tissue injury in kidney disease and KTx along with the types of stem therapies that have been developed to address them. (b) Critically evaluate the extent to which clinical trials of stem cell products have validated such effects in humans with kidney disease and KTx. (c) Identify key bottlenecks to the large-scale application of stem cell therapies to reduce the burden of kidney disease on patients and societies.
Muscle satellite cells are adult muscle stem cells indispensable for growth and regeneration of postnatal skeletal muscle. Notch plays a central role in maintenance of muscle satellite cells, but how Notch maintains the muscle stem cell pool is not fully understood. Previously, we reported that a prostaglandin E2 receptor, EP2, is upregulated by Notch signal and suppresses differentiation of human muscle progenitors. Here we examined the roles of EP2 in muscle satellite cells using a mouse Cre-LoxP conditional gene knockout system. Genetic inactivation of the EP2 gene (PTGER2) activated muscle satellite cells, caused their loss, and impaired muscle regeneration. These results indicate that EP2 is indispensable for maintenance of satellite cells. Ex vivo analysis using isolated myofibers showed that prostaglandin E2 (PGE2) delayed the activation of satellite cells via EP2. An extracellular signal-regulated kinase (ERK) 1/2 inhibitor blocked the activation of satellite cells on myofibers, and PGE2 attenuated the phosphorylation of ERK1/2 in muscle satellite cells. These results suggest that EP2 keeps the quiescence of satellite cells and maintains the satellite cell pool in part by inhibiting the ERK1/2 signaling pathway.
Breast cancer (BC) has a high incidence and mortality rate among women. Doxorubicin (DOX) is one of the standard chemotherapeutic drugs for BC. However, chemoresistance to DOX represents a major therapeutic obstacle, and its underlying mechanisms remain elusive. CircEGFR (hsa_circ_0080222) is significantly upregulated in BC tissues and drives the malignant progression of BC. However, the role of circEGFR in DOX resistance remains unclear. In this study, we found that circEGFR expression was significantly upregulated in DOX-resistant MCF-7 (MCF-7/DOX) cells. CircEGFR overexpression attenuated DOX sensitivity in MCF-7 cells, as evidenced by increased cell viability and enhanced proliferative capacity. Notably, circEGFR also enhanced cancer stem cell (CSC) properties in BC cells. Conversely, circEGFR knockdown reduced chemoresistance and stem-like properties in both MDA-MB-231 and MCF-7/DOX cells. Mechanistically, circEGFR interacted with IGF2BP2 to enhance the stability of the stem cell marker SOX2 mRNA, resulting in elevated SOX2 expression. Rescue experiments demonstrated that silencing IGF2BP2 or SOX2 abrogated circEGFR-mediated chemoresistance and stemness properties in BC cells. In conclusion, our study demonstrates for the first time that circEGFR drives DOX chemoresistance and maintains cancer stemness in BC cells via the IGF2BP2/SOX2 axis. Targeting circEGFR may be considered a promising therapeutic strategy for overcoming chemotherapy resistance in BC.
Stem cells use oxidized nicotinamide adenine dinucleotide (NAD+) in distinct subcellular compartments to support self-renewal and to regulate chromatin. There is limited information, however, about the biosynthetic pathways that replenish intracellular NAD+, which is continuously turned over in undifferentiated mouse embryonic stem cells. Establishing specific metabolic inputs for maintaining self-renewal can help direct reprogramming efforts. We used single fluorescent protein biosensors for in situ NAD+ measurements in J1 mouse embryonic stem cells. Sensors and controls were localized to the nucleus, cytoplasm, and mitochondrial compartments. Using a specific inhibitor for nicotinamide salvage, we found that loss of this pathway depleted NAD+ concentrations in all three subcellular compartments in undifferentiated culture conditions. We determined that loss of nicotinamide salvage reduced colony size, extended cell cycle, and resulted in diminished expression of self-renewal markers. Supplementation with precursors in the nicotinamide salvage pathway bypassed the pharmacological block, replenished cytosolic NAD+ levels, and reversed the effects on colony size. Notably, supplementation with deaminated precursors did not replenish intracellular NAD+ levels, suggesting minimal contribution from this pathway at this stage. In support, expression data from multiple mouse and human lines showed that nicotinamide salvage pathway enzyme NAMPT was predominantly expressed at the embryonic stem cell stage compared to the enzymes in other NAD+ biosynthesis pathways. Collectively, the data showed that undifferentiated embryonic stem cells heavily rely on nicotinamide salvage, indicating that this dependency is conserved.
Hematopoietic stem cell (HSC) transplantation is a potentially curative option for patients with hematologic malignancies, but donor shortages impact graft availability. Umbilical cord blood (UCB) is a viable alternative source of HSC; however, the limited numbers present in a single unit have spurred efforts to expand HSC ex vivo. We previously demonstrated that the addition of valproic acid (VPA), an anti-convulsive drug, to CB cell cultures promotes maintenance of functional HSC, but not expansion. However, it has been proposed that VPA primarily induces mitochondrial reprogramming of mature CD34+CD90- cells to more primitive CD34+CD90+ cells, rather than the replication of CD34+CD90+ cells in culture. To determine which fraction of the CD34+CD90+ cells present after culture in VPA were derived from CD34+CD90- vs. CD34+CD90+ cells, we examined the functionality of CD34+CD90+ cells derived from each flow cytometry-sorted population. During culture in VPA there was a significant increase in CD34+CD90+ cell number; the majority arising from pre-existing CD34+CD90+ cells, with minimal contribution from CD34+CD90- cells. Colony-forming unit (CFU) assays revealed reduced plating efficiency and xeno-transplantation studies demonstrated diminished in vivo hematopoietic reconstitution potential of CD34+CD90+ cells derived from relatively committed CD34+CD90- cells. Our findings indicate that while VPA supports CD34+CD90+ cell expansion, the CD34+CD90+ cells derived from CD34+CD90- cells are functionally more differentiated than those derived directly from CD34+CD90+ cells, with increased mitochondrial mass and membrane potential, but reduced regenerative potential. These results emphasize the need for functional assessments of culture-expanded HSCs to accurately determine their therapeutic potential.
Initially regarded as insignificant cellular waste, extracellular vesicles (EVs) are now recognized as key mediators of intercellular communication, capable of transferring bioactive molecules-such as proteins, nucleic acids, and small compounds-between cells. This function has positioned EVs as promising cell-free therapeutic agents with the potential to transform modern medicine. Stem and progenitor cells naturally release EVs that can replicate many of the therapeutic effects of cell transplantation, while avoiding the challenges associated with administering living cells. EVs derived from various cell sources-including embryonic stem cells, induced pluripotent stem cells, mesenchymal stem cells, stromal cells, cardiac progenitor cells, and endothelial progenitor cells-have shown therapeutic efficacy in preclinical models of ischemic heart disease. EVs' translation into human trials in cardiac therapy has lagged behind, however, largely due to challenges related to EV production standardization, regulatory frameworks, and the demonstration of reproducible efficacy in human subjects. Nevertheless, recent milestones have been achieved with the successful completion of the phase I EV-AMI trial (Safety Evaluation of Intracoronary Infusion of EVs in Patients with Acute Myocardial Infarction, NCT04327635) and the enrollment of the first patient in the phase I SECRET-HF trial (Treatment of Non-ischemic Cardiomyopathies by Intravenous Extracellular Vesicles of Cardiovascular Progenitor Cells, NCT05774509). This concise review outlines the evolution of EVs from basic biological discovery to innovative therapeutic platforms, with particular emphasis on their potential applications in acute myocardial infarction. Remaining challenges for clinical translation, including manufacturing and regulatory hurdles, will also be discussed.
The precise timing of stem cell specification and niche formation during murine incisor development is poorly understood, and it is unclear whether these processes occur simultaneously or in a sequential manner. Functional dental epithelial stem cells are marked by the expression of Sox2, a transcription factor that is broadly expressed in the dental epithelium at the dentition onset and restricted to stem cells in fully developed incisor. Using genetic lineage tracing in Sox2CreERT2/+; R26RmT/mG and Sox2CreERT2/+; R26RtdT/+ embryos along with a single-cell RNA sequencing at different stages of incisor development, we investigated the timing of the stem cell specification and its temporal relationship with niche formation. Our results reveal the presence of a Sox2-expressing stem cell-like population prior to formation of the functional niche. These cells localize to the leading edge of the advancing incisor epithelium where they are maintained in an undifferentiated state. Our data demonstrate presence of actomyosin network and a generation of a contractile tension, which helps confine Sox2+ stem cells to the leading edge. This mechanical confinement likely plays an important role in maintaining their stemness until the niche is functionally and structurally established. Partial or complete disruption of the actomyosin network disables the clustering of Sox2-expressing cells, potentially triggering their premature differentiation, and ultimately leads to impaired formation of the functional stem cell niche and abnormal growth of the incisor.
To explore the effect of exosomes-encapsulated miR3671 (EXO-miR3671) from mesenchymal stem cells (MSC) on diabetic wound healing and its mechanism. Bioinformatics analysis was conducted to identify gene and miRNA expression changes associated with diabetic foot ulcer (DFU). A diabetic mouse wound model was established, and dual-luciferase reporter assay was performed to verify the targeting relationship between miR3671 and QSOX1. For in vitro experiments, human umbilical vein endothelial cells (HUVECs) were cultured and transfected; the effects of EXO-miR3671 secreted by mesenchymal stem cells on HUVEC biological functions were detected via EdU proliferation assay, Transwell migration assay, and angiogenesis assay. For in vivo experiments, after dynamic measurement of wound areas in diabetic mice, HE staining, immunohistochemistry, and Western blot were employed to evaluate the regulatory effects of mesenchymal stem cell-derived EXO-miR3671 on wound healing process and related protein expression levels. MiR3671 was downregulated in DFU tissues and diabetic mouse wounds. EXO-miR3671 promoted HUVEC proliferation, migration, and angiogenesis in vitro. In diabetic mice, it accelerated wound closure, reduced inflammation, increased Ki67+ cell proliferation, and upregulated pro-angiogenic growth factors. MSC-derived EXO-miR3671 could be a potential treatment for diabetic wounds, providing new insights for treatment.
Despite breakthroughs in molecularly targeted and immune therapies, the prognosis for patients with urinary bladder cancer (UBC) has remained unsatisfactory over the past few decades. Understanding the molecular underpinnings of UBC treatment refractoriness is crucial for identifying novel therapeutic targets and strategies. Cancer stemness plays a pivotal role in the oncogenesis and treatment resistance of UBC, while the underlying molecular regulatory mechanisms are poorly understood. We identified isoform 1 of ASPM (ASPM-i1) as the most upregulated stemness-associated factor in tumorigenic UBC cells, predominantly expressed by ALDH1+ stem-like cancer cells. Pairing genetic ASPM-i1 inhibition with standard chemotherapeutic agents used in the treatment of UBC, including cisplatin and gemcitabine, circumvents the treatment resistance of tumorigenic and stem-like UBC cells. Mechanistically, ASPM-i1 interacts with the Disheveled (DVL) and intracellular NOTCH proteins, thereby attenuating CUL3- or FBXW7-mediated ubiquitination and the subsequent proteasomal degradation. The regulatory module concomitantly enhances the activities and ligand responsiveness of the Wnt and Notch signaling pathways in UBC cells. As a result, ASPM-i1 inhibition sensitized tumorigenic UBC cells to chemotherapy in a NOTCH- and DVL-dependent manner. In human UBC tissues, ASPM-i1 shows substantial cell-to-cell heterogeneity and is upregulated in a subset (46.4%) of tumors, correlating with poor clinical prognosis. This study reveals a crucial co-regulatory module of Notch and Wnt signaling that mediates stemness and chemotherapy resistance in tumorigenic UBC cells; its inhibition provides a novel approach to enhance chemosensitivity and improve therapeutic outcomes in human UBC.
Huntington's disease (HD) is a neurodegenerative disorder caused by cytosine-adenine-guanine (CAG) triplet expansion in the HTT gene, producing a mutant Huntingtin protein that impairs mitochondrial dynamics by reducing fusion and increasing fission. Mesenchymal stem cells (MSCs) have shown potential therapeutic effects by sharing functional mitochondria and other secretomes. In this study, quinolinic acid-lesioned neuro-2a (QA-N2a) cells and glutamatergic neurons with 50 CAG repeats (HD neurons) were co-cultured with human umbilical cord-derived MSCs for 5 hours. For QA-N2a cells, immunocytochemistry (ICC) was performed to demonstrate the change in GABA and Substance P before and after co-culture. For HD neurons, ICC was conducted to identify mitochondrial proteins, while Western blot was employed to evaluate proteins related to inflammation and mitochondrial function. As a result, co-culture with MSC significantly restored the expression of GABA and Substance P, which diminished after QA exposure. In HD neurons co-cultured with MSCs, an increase in mitochondrial abundance was observed, with significantly higher intensity and dendritic distribution of mitochondria compared to control cells. Western blot analysis confirmed this increase and showed a rising trend in ATP5a levels. MSCs also promoted mitochondrial fusion, indicated by higher levels of Mitofusin 2 (MFN2) and Mitochondrial Dynamin Like GTPase (OPA1), and a trend of reduction in the fission marker Dynamin-Related Protein (DRP1). Additionally, the co-culture led to a decreased trend in neuroinflammation markers IL-6, TNF-α, MMP9, and p-NFkB. Collectively, this study demonstrates that MSCs alleviate HD pathology by restoring the mitochondria activity and potentially suppressing inflammation in two different HD in vitro models.
Hematopoietic stem cell (HSC) transplantation is a lifesaving therapy for hematologic diseases, but its broader application remains constrained by challenges in sourcing, manipulating, and reliably expanding functional HSCs. In this review, we discuss strategies to expand and engineer HSCs by recreating essential aspects of the bone marrow niche. These include defined cytokine cocktails, small molecule modulators, stromal co-culture systems, and biomaterials that promote self-renewal while limiting differentiation. We highlight advances in three-dimensional organoid models and microfluidic platforms that better support long-term repopulating cells and reflect native microenvironments. In parallel, progress in gene delivery platforms, including both viral and nonviral approaches, is enabling more efficient and targeted modification of HSCs for therapeutic use in genetic disorders such as sickle cell disease and β-thalassemia. While these tools have advanced significantly, significant hurdles remain in scaling, preserving stem cell identity, and reducing culture-induced stress. Continued refinement of biomimetic systems and genome engineering technologies will be central to expanding the clinical utility of HSC-based therapies.
Rodent and primate digit tips exhibit a remarkable regenerative capacity following amputation, driven by highly proliferative nail stem cells (NSCs) with active canonical Wnt signaling. Recently, a distinct, slow-cycling population of bi-functional nail proximal fold stem cells (NPFSCs) has been identified, contributing to both peri-nail epidermis and nail plate (NP). Here, we demonstrate that NPFSCs actively participate in nail growth, orchestrating digit regeneration, with BMP signaling serving as a key regulator. Inhibition of BMP resulted in an epidermalized NP-like structure with limited regeneration due to impaired Wnt pathway activation in the nail matrix cells. Conversely, BMP activation enhanced NPFSCs' involvement in the nail matrix and significantly promoted digit regeneration. We further revealed that enhanced BMP activity not only accelerated nail and bone regrowth but also extended the regenerative boundary proximally, enabling full regeneration after up to ∼60% removal of the distal phalanx (P3). Moreover, in BMP gain-of-function (GoF) models, extreme proximal amputation, removing the majority of the P3, still permitted partial NP regeneration despite the absence of bone reconstruction. Finally, we isolated and cultured lineage-traced NPFSCs and transplanted them into immunocompromised mice, where they integrated into the nail proximal fold and contributed to nail matrix progenitors during regeneration. Transplanted NPFSCs retained their regenerative capacity in vivo, highlighting their therapeutic potential. Collectively, our findings identify a pivotal role of BMP signaling in mediating NPFSC-driven digit regeneration, reveal BMP-Wnt cross-talk as essential to this process, and provide a framework for enhancing regenerative outcomes in previously non-regenerative contexts following traumatic amputation.
Diabetic enteropathy (DE) is a common complication of diabetes mellitus (DM), yet its underlying molecular mechanisms remain poorly understood. Emerging evidence suggests that abnormal differentiation of intestinal epithelial stem cells (IESCs) contributes to early intestinal dysfunction in DM. In this study, we aimed to investigate the role of CircVapa in regulating IESC differentiation and to elucidate the underlying molecular mechanism involving the miR-212-3p/Smoc2 axis. In this study, IESCs were extracted from BKS.CgDock7m+/+Lepr db/JNju (DM) mice models. A circular RNA molecule, CircVapa, was identified as being markedly enriched in IESCs. Experimental suppression of CircVapa in diabetic mice attenuated abnormal differentiation of intestinal epithelial cells (IECs). Notably, CircVapa was identified as a critical mediator of Lgr5+ stem cell functionality in hyperglycemic conditions. Mechanistically, microarray analysis, bioinformatics analysis, and luciferase reporter assays demonstrated that CircVapa serves as a competitive endogenous RNA (ceRNA) by directly binding miR-212-3p, thereby regulating Smoc2 expression. Furthermore, CircVapa regulated abnormal IESC differentiation via the miR-212-3p/Smoc2 regulatory network in diabetic mice. Collectively, this study demonstrates an important role of CircVapa in regulating IEC differentiation during DM progression.
End-stage renal disease (ESRD) is a major global health burden, and current treatments, such as dialysis and kidney transplantation, remain constrained by donor shortages, procedure-related complications, and reduced long-term quality of life. Regenerative medicine, particularly stem cell-based approaches, offers promising next-generation strategies for kidney repair and replacement. This review summarizes the current understanding of kidney development and intrinsic regenerative capacity and evaluates the therapeutic potential of hematopoietic stem cells, mesenchymal stem cells (MSCs), kidney-derived stem cells, and induced pluripotent stem cell (iPSC)-derived kidney organoids. Evidence from preclinical models demonstrates renoprotective and immunomodulatory effects across multiple stem cell types, whereas early-phase clinical trials have reported favorable safety profiles and preliminary signals of the efficacy of MSC-based therapies. iPSC- and organoid-based approaches present additional challenges, including incomplete vascularization, immature nephron structures, risks of tumorigenicity, immune compatibility issues, and the need for reproducible good manufacturing practice (GMP)-compliant manufacturing. Advances in biomaterials, organoid engineering, and vascularization strategies may help overcome these barriers. Overall, stem cell-based regenerative therapies show substantial potential to complement or ultimately reduce the reliance on dialysis and transplantation. Continued technological innovations and rigorously designed clinical trials are critical to translate these promising approaches into clinical practice.
Abnormal decidual natural killer cell (dNK) function is linked to pregnancy complications occurring in both early and late gestation, including recurrent pregnancy loss, preeclampsia, and preterm birth. Exploration of dNK heterogeneity as it relates to function is an active area of research; however, most of this work has focused on early gestation. Using flow cytometric and transcriptomic single-cell definitions of dNK subtypes, we characterized dNK heterogeneity in term dNK within both chorioamniotic membranes and the basal plate. We also applied aptamer-based secretome profiling to first trimester and term dNK and found dNK-specific proteins-VEGF and PLGF-to be reduced at term. We further determined that, compared to first trimester dNK, term dNK have reduced cytotoxicity against target cells. Finally, we applied this knowledge to establish a protocol for differentiation of induced pluripotent stem cells (iPSC) into functional dNK. We found that treatment with TGFβ enriched for dNK2 subtype, while inducing dNK markers, CD9 and CD103. We evaluated function using cytokine and degranulation assays, aptamer-based secretome profiling, and cytotoxicity assays. We found that iPSC-dNK are functionally most similar to primary dNK. Further, TGFβ iPSC-dNK had reduced GM-CSF in response to PMA/I and increased secretion of VEGF and other first trimester-specific proteins-supportive of a shift towards an early gestation, dNK2-dominant, phenotype. We conclude that changes in dNK function across gestation reflect shifts in dNK subtypes that can be reproducibly derived from iPSC, providing a new method for modeling dNK and laying the foundation for cell-based therapeutics for reproductive disease.
Catechin (CH) exhibits protective effects on bone metabolism, but its underlying mechanism remains incompletely understood. We investigated the osteogenic effects of CH and its molecular pathways using bone marrow mesenchymal stem cells and MC-3T3-E1 preosteoblasts. Cell viability was assessed after CH treatment (1-100 μg/mL). Osteogenic differentiation was evaluated by ALP activity, mineralization, and the expression of key markers (Runx2, Opn, Ocn, Sp7). Mechanistic studies involved examining autophagy markers (LC3-II, P62) and the AMPK pathway, using pharmacological inhibitors (compound C for AMPK; 3-methyladenine for autophagy). The protective role of CH under oxidative stress was tested in hydrogen peroxide-treated cells by measuring viability, ROS levels, NRF2 translocation, and osteogenic capacity. CH showed no significant cytotoxicity up to 100 μg/mL. At 10 μg/mL, it significantly enhanced osteogenic differentiation, increasing alkaline phosphatase activity (ALP), mineralization, and the gene/protein levels of osteogenic markers. CH activated autophagy (elevated LC3-II, decreased P62) and the AMPK pathway. Inhibition of AMPK or autophagy partially suppressed CH-induced osteogenesis, which was significantly rescued by CH co-treatment. Under oxidative stress, CH improved cell viability, reduced intracellular ROS, inhibited NRF2 nuclear translocation, and restored osteogenic differentiation. CH promotes osteogenesis primarily via the AMPK-autophagy axis and reverses oxidative stress-induced suppression of osteogenic differentiation through ROS clearance. These findings highlight its therapeutic potential for bone regeneration and related disorders.
Peripheral nerve injuries (PNIs) present a persistent clinical challenge due to the intrinsically limited regenerative capacity of peripheral nerves. While dental pulp stem cells (DPSCs) exhibit significant neuroregenerative potential, their therapeutic efficacy is constrained by hostile microenvironments and inherent functional heterogeneity. Genetic modification may offer a promising strategy to enhance their therapeutic capabilities. DPSCs were induced toward neural lineage differentiation, and key gene candidates were identified through qRT-PCR. Lentiviral-mediated gene interference was performed to modulate target gene expression, followed by comprehensive analysis of differentiation outcomes using qRT-PCR, Western blotting, and immunofluorescence assays. RNA sequencing was employed to uncover associated signaling pathways, which were subsequently validated through pharmacological inhibition with specific inhibitors. The therapeutic efficacy of genetically engineered DPSCs was evaluated in a rat model of sciatic nerve crush injury, with neural regeneration quantitatively assessed via neuroelectrophysiological measurements and histological analyses. LARP7 positively regulated the Schwann cell-like differentiation of DPSCs, as well as their trophic and anti-inflammatory effects, thus enhancing its therapeutic effects on nerve repair and promoting functional recovery. Mechanistically, we found that LARP7 remodeled cytokine-cytokine receptor interactions, enhancing trophic support while attenuating proinflammatory responses, and activated the PI3K-Akt-mTOR signaling pathway, with ERBB4 serving as a critical downstream effector, promoting DPSCs differentiation into Schwann cell-like phenotypes. Collectively, LARP7-mediated changes in DPSCs establish a new therapeutic paradigm that addresses the limitations of current stem cell-based interventions and enables the development of standardized biotherapeutics for peripheral nerve repair.
Sclerotomal progenitors derived from pluripotent stem cells hold promises for modeling skeletal development and recapitulating the perinatal marrow niche that may provide insights into hematopoietic niche formation and immune regulation. Current strategy to derive mouse sclerotomal progenitors suffered from low differentiation efficiency and heterogeneous cell progenies. Here, we developed a fast and efficient strategy to generate sclerotomal progenitors by accelerated induction from primitive streak (PS) through modulating BMP and SHH signaling, achieving an 86.9% differentiation efficiency. Moreover, the resulting progenitors showed similar global gene expression profiles to those of the primary sclerotome, possessed strong osteochondral bipotential, and could recapitulate key features of endochondral ossification upon micromass-mediated osteogenic induction, including perinatal bone marrow (BM)-like niches regeneration. Our findings underscore accelerated sclerotomal induction from the primitive streak as an efficient strategy to derive sclerotomal progenitors for skeletal modeling and BM niche bioengineering.