Stem Cell Research & Therapy最新文献

{"title":"Extracellular vesicles from hiPSC-derived NSCs protect human neurons against Aβ-42 oligomers induced neurodegeneration, mitochondrial dysfunction and tau phosphorylation.","authors":"Shama Rao, Leelavathi N Madhu, Roshni Sara Babu, Goutham Shankar, Sanya Kotian, Advaidhaa Nagarajan, Raghavendra Upadhya, Esha Narvekar, James J Cai, Ashok K Shetty","doi":"10.1186/s13287-025-04324-3","DOIUrl":"https://doi.org/10.1186/s13287-025-04324-3","url":null,"abstract":"<p><strong>Background: </strong>Alzheimer's disease (AD) is characterized by the accumulation of amyloid beta-42 (Aβ-42) in the brain, causing various adverse effects. Thus, therapies that reduce Aβ-42 toxicity in AD are of great interest. One promising approach is to use extracellular vesicles from human induced pluripotent stem cell-derived neural stem cells (hiPSC-NSC-EVs) because they carry multiple therapeutic miRNAs and proteins capable of protecting neurons against Aβ-42-induced toxicity. Therefore, this in vitro study investigated the proficiency of hiPSC-NSC-EVs to protect human neurons from Aβ-42 oligomers (Aβ-42o) induced neurodegeneration.</p><p><strong>Methods: </strong>We isolated hiPSC-NSC-EVs using chromatographic methods and characterized their size, ultrastructure, expression of EV-specific markers and proficiency in getting incorporated into mature human neurons. Next, mature human neurons differentiated from two different hiPSC lines were exposed to 1 µM Aβ-42o alone or with varying concentrations of hiPSC-NSC-EVs. The protective effects of hiPSC-NSC-EVs against Aβ-42o-induced neurodegeneration, oxidative stress, mitochondrial dysfunction, impaired autophagy, and tau phosphorylation were ascertained using multiple measures and one-way ANOVA with Newman-Keuls multiple comparisons post hoc tests.</p><p><strong>Results: </strong>A significant neurodegeneration was observed when human neurons were exposed to Aβ-42o alone. Neurodegeneration was associated with (1) elevated levels of reactive oxygen species (ROS), mitochondrial superoxide, malondialdehyde (MDA) and protein carbonyls (PCs), (2) increased expression of proapoptotic Bax and Bad genes and proteins, and genes encoding mitochondrial complex proteins, (3) diminished mitochondrial membrane potential and mitochondria, (4) reduced expression of the antiapoptotic gene and protein Bcl-2, and autophagy-related proteins, and (5) increased phosphorylation of tau. However, the addition of an optimal dose of hiPSC-NSC-EVs (6 × 10⁹ EVs) to human neuronal cultures exposed to Aβ-42o significantly reduced the extent of neurodegeneration, along with diminished levels of ROS, superoxide, MDA and PCs, normalized expressions of Bax, Bad, and Bcl-2, and autophagy-related proteins, higher mitochondrial membrane potential and mitochondria, enhanced expression of genes linked to mitochondrial complex proteins, and reduced tau phosphorylation.</p><p><strong>Conclusions: </strong>An optimal dose of hiPSC-NSC-EVs could significantly decrease the degeneration of human neurons induced by Aβ-42o. The results support further research into the effectiveness of hiPSC-NSC-EVs in AD, particularly their proficiency in preserving neurons and slowing disease progression.</p>","PeriodicalId":21876,"journal":{"name":"Stem Cell Research & Therapy","volume":"16 1","pages":"191"},"PeriodicalIF":7.1,"publicationDate":"2025-04-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12008877/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144046552","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The role of PKM2-mediated metabolic reprogramming in the osteogenic differentiation of BMSCs under diabetic periodontitis conditions.","authors":"Yanlin Zhu, Yuhan Yang, Yuyan Lan, Zun Yang, Xiang Gao, Jie Zhou","doi":"10.1186/s13287-025-04301-w","DOIUrl":"https://doi.org/10.1186/s13287-025-04301-w","url":null,"abstract":"<p><strong>Background: </strong>Diabetes mellitus (DM) and periodontitis have a bidirectional relationship, with each being a high-risk factor for the other. Prolonged hyperglycemia exacerbates periodontal inflammation and disrupts bone homeostasis. Pyruvate kinase M2 (PKM2), a key enzyme in glycolysis, is involved in metabolic reprogramming, but its role in osteogenesis under high-glucose (HG) inflammatory conditions remains largely unknown. This study aimed to investigate the effects of HG and inflammation on bone marrow mesenchymal stem cells (BMSCs) under indirect co-culture conditions and to explore how PKM2 regulates metabolism and mitochondrial function during osteogenic differentiation in HG inflammatory environments, elucidating its role in diabetic periodontitis (DP).</p><p><strong>Methods: </strong>Expose BMSCs to conditioned medium (CM) collected from RAW264.7 cells stimulated with HG and/or lipopolysaccharide (LPS). BMSCs functionality was assessed using CCK8, EdU, Annexin V-PI apoptosis assay, alkaline phosphatase (ALP), and Alizarin Red S (ARS) staining. Metabolic characteristics were evaluated through Seahorse assays, lactate production, glucose uptake, and ATP measurements. Mitochondrial function was assessed via JC-1, and ROS staining, Mito-Tracker staining, and transmission electron microscopy (TEM). Gene and protein expression were analyzed by quantitative real-time PCR and western blotting. In vivo therapeutic effects of shikonin were validated via micro-CT and histological staining in a diabetic periodontitis mouse model.</p><p><strong>Results: </strong>In vitro experiments demonstrated that HG inflammatory conditions impaired the survival of BMSCs, suppressed osteogenic differentiation, and induced metabolic reprogramming. This reprogramming was characterized by enhanced glycolysis, impaired oxidative phosphorylation (OXPHOS), abnormal upregulation of PKM2 expression, and mitochondrial dysfunction accompanied by morphological alterations. Shikonin effectively reversed these adverse effects by inhibiting PKM2 tetramerization, rescuing the loss of osteogenic function in BMSCs. The therapeutic potential of shikonin was confirmed in the diabetic periodontitis mouse model.</p><p><strong>Conclusion: </strong>PKM2 impairs the osteogenesis of BMSCs by affecting metabolism and mitochondrial function, suggesting it as a potential therapeutic target for diabetic periodontitis.</p>","PeriodicalId":21876,"journal":{"name":"Stem Cell Research & Therapy","volume":"16 1","pages":"186"},"PeriodicalIF":7.1,"publicationDate":"2025-04-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12008901/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143986655","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Chemical reprogramming culture for the expansion of salivary gland epithelial basal progenitor cells.","authors":"Ye Jin Jeong, Yongpyo Hong, Yeo-Jun Yoon, Nam Suk Sim, Seung-Min Hong, Jae-Yol Lim","doi":"10.1186/s13287-025-04295-5","DOIUrl":"https://doi.org/10.1186/s13287-025-04295-5","url":null,"abstract":"<p><strong>Background: </strong>Salivary gland (SG) hypofunction presents a significant clinical challenge with limited treatment options. SG epithelial cells offer a promising approach due to their intrinsic tissue specificity and regenerative potential. However, the lack of efficient culture methods has hindered their clinical use.</p><p><strong>Methods: </strong>This study presents a chemical reprogramming culture (CRC) system that utilizes a combination of three small molecules for the long-term two-dimensional culture of human SG epithelial progenitor cells. We characterized the cultured cells, measured their organoid-forming efficiencies, and assessed their differentiation potential. To evaluate the therapeutic efficacy of the SG basal progenitor cells (SG-BPCs), we administered them into a mouse model with radiation-induced SG hypofunction and assessed the functional recovery.</p><p><strong>Results: </strong>By utilizing optimal concentrations of the small molecules Y-27632, A83-01, and LDN193189, the SG epithelial cells achieved over 50 population doubling levels (PD) within 80 d, surpassing the Hayflick limit. β-galactosidase and Terminal deoxynucleotidyl transferase dUTP nick end labeling staining confirmed that these small molecules inhibited cellular senescence and apoptosis, respectively. The cells expressed SG basal ductal cell markers KRT5, KRT19, and SOX9, with increased expression levels observed from PD5 to PD40. Notably, these expanded cells were able to differentiate into various SG cell types, including acinar and myoepithelial cells, indicating that SG-basal progenitor cells (SG-BPCs) were selectively proliferated using our CRC method. To assess the therapeutic potential of the expanded SG-BPCs, they were administered to mice with radiation-induced SG hypofunction. The treatment successfully restored SG function.</p><p><strong>Conclusion: </strong>Our findings demonstrate that our CRC system is an effective method for the long-term culture of SG-BPCs. This advancement holds significant promise for the development of SG epithelial progenitor-based therapies to treat SG hypofunction.</p>","PeriodicalId":21876,"journal":{"name":"Stem Cell Research & Therapy","volume":"16 1","pages":"187"},"PeriodicalIF":7.1,"publicationDate":"2025-04-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12008940/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144012614","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The Immunomodulatory mechanism and research progress of mesenchymal stem cells in the treatment of allergic rhinitis.","authors":"Yu-Meng Ye, Yu-Xin Zhao, Li-Rong Xiang, Chen-Yu Zou, Hao Xiao, Huan Lu, Hui Yang, Juan-Juan Hu, Hui-Qi Xie","doi":"10.1186/s13287-025-04333-2","DOIUrl":"https://doi.org/10.1186/s13287-025-04333-2","url":null,"abstract":"<p><strong>Background: </strong>Allergic rhinitis (AR) affects 10-40% of the global population, yet current therapies (drugs, immunotherapy) face limitations in efficacy and safety. Mesenchymal stem cells (MSCs) have emerged as a promising alternative due to their immunomodulatory properties.</p><p><strong>Key findings: </strong>Preclinical studies demonstrate that MSCs from adipose, bone marrow, umbilical cord, and tonsils reduce AR symptoms (sneezing, nasal inflammation) and serum IgE (Immunoglobulin E) levels by restoring Th1/Th2 immune equilibrium and enhancing Treg (Regulatory T cells) activity. MSC-derived exosomes and hydrogel-encapsulated formulations further improve targeting and safety. However, clinical translation is hindered by heterogeneous protocols and unresolved long-term risks (e.g., tumorigenicity).</p><p><strong>Clinical significance: </strong>MSC-based therapies offer potential for durable AR remission by addressing immune dysregulation at its root. Future efforts must prioritize standardized production, phase I safety trials, and combination strategies (e.g., exosomes + hydrogels) to accelerate clinical adoption.</p>","PeriodicalId":21876,"journal":{"name":"Stem Cell Research & Therapy","volume":"16 1","pages":"188"},"PeriodicalIF":7.1,"publicationDate":"2025-04-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12008879/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144062175","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Inhibition of ceramide synthesis improves the outcome of ischemia/reperfusion injury in cardiomyocytes derived from human induced pluripotent stem cell.","authors":"Pellumb Haxhikadrija, Jasmine M F Wu, Sascha Hübner, Katja Grün, Tom Kretzschmar, Tina Müller, Markus H Gräler, Claudia Backsch, Anja Weise, Elisabeth Klein, P Christian Schulze, Mohamed M Bekhite","doi":"10.1186/s13287-025-04340-3","DOIUrl":"https://doi.org/10.1186/s13287-025-04340-3","url":null,"abstract":"<p><strong>Background: </strong>Ceramides are bioactive sphingolipids that have physiological effects on inflammation, apoptosis, and mitochondrial dysfunction. They may play a critical role in the harm of ischemia/reperfusion (IR). Ceramides and IR injury are not well-studied, and there is a lack of human data.</p><p><strong>Methods and results: </strong>Current studies aimed to investigate the role of ceramide buildup in cardiomyocytes (CMs) death using CMs derived from human induced pluripotent stem cell (hiPSC) as a model for simulating IR injury in vitro. In our model, serum- and glucose-free media was used to expose hiPSC-derived CMs to hypoxia (3% O₂) for 6 h (hrs), followed by reoxygenation (20% O₂) for 16 h. In contrast to normoxia (control) or hypoxia (ischemia), our data showed that following IR, there was an increase in the formation of mitochondrial superoxide and the mRNA levels of genes regulating ceramide synthesis, such as CerS2 and CerS4 in CMs. Further, there was a considerable rise in the levels of total ceramide, long-chain (C16:0, C18:0, and C18:1), and very long-chain (C22:0 and C24:1) ceramide species in CMs following reperfusion in comparison to control or ischemic CMs. Interestingly, compared to CMs exposed to IR without inhibitor, our data showed that inhibition of ceramide formation with fumonisin B1 (FB1) significantly lowered ceramide levels, reduced apoptosis, improved mitochondrial function, and enhanced survival of CMs exposed to IR. Furthermore, we used a transgenic mouse model, in which the CerS2 gene was overexpressed in the CMs of α-MHC-CerS2 mice, to validate the basic idea that ceramide contributes to heart disease in vivo. Our results showed that the heart tissues of α-MHC-CerS2 mice had significant levels of long-chain and very long-chain ceramides, which causes increased apoptosis, proinflammatory cytokines, interstitial inflammatory cell infiltration, and collagen deposition.</p><p><strong>Conclusions: </strong>Results from both in vitro and in vivo experiments show that ceramides have a significant role in either mediating or inducing damage to CMs. Additionally, in vitro findings show that ceramide reduction improves the outcome of IR injury by lowering intracellular Ca²⁺ [Ca²⁺]_i concentration and improves mitochondrial function changes during IR.</p>","PeriodicalId":21876,"journal":{"name":"Stem Cell Research & Therapy","volume":"16 1","pages":"190"},"PeriodicalIF":7.1,"publicationDate":"2025-04-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12008854/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144054183","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Correction: Targeting stem cell niche can protect hematopoietic stem cells from chemotherapy and G-CSF treatment.","authors":"Sidan Li, Dehui Zou, Changhong Li, Hengxing Meng, Weiwei Sui, Sizhou Feng, Tao Cheng, Qiongli Zhai, Lugui Qiu","doi":"10.1186/s13287-025-04336-z","DOIUrl":"https://doi.org/10.1186/s13287-025-04336-z","url":null,"abstract":"","PeriodicalId":21876,"journal":{"name":"Stem Cell Research & Therapy","volume":"16 1","pages":"185"},"PeriodicalIF":7.1,"publicationDate":"2025-04-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12007240/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144032185","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Harnessing stem cell-derived exosomes: a promising cell-free approach for spinal cord injury.","authors":"Miaoman Lin, Farzaneh Alimerzaloo, Xingjin Wang, Obada Alhalabi, Sandro M Krieg, Thomas Skutella, Alexander Younsi","doi":"10.1186/s13287-025-04296-4","DOIUrl":"https://doi.org/10.1186/s13287-025-04296-4","url":null,"abstract":"<p><p>Spinal cord injury (SCI) is a severe injury to the central nervous system that often results in permanent neurological dysfunction. Current treatments have limited efficacy and face challenges in restoring neurological function after injury. Recently, stem cell-derived exosomes have gained attention as an experimental treatment for SCI due to their unique properties, including superior biocompatibility, minimal immunogenicity and non-tumorigenicity. With their potential as a cell-free therapy, exosomes promote SCI repair by enhancing nerve regeneration, reducing inflammation and stabilizing the blood-spinal cord barrier. This review summarizes advances in stem cell-derived exosome research for SCI over the past years, focusing on their mechanisms and future prospects. Despite their promising therapeutic potential, clinical translation remains challenging due to standardization of exosome isolation protocols, compositional consistency and long-term safety profiles that require further investigation.</p>","PeriodicalId":21876,"journal":{"name":"Stem Cell Research & Therapy","volume":"16 1","pages":"182"},"PeriodicalIF":7.1,"publicationDate":"2025-04-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12004558/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144039453","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Small extracellular vesicles: the origins, current status, future prospects, and applications.","authors":"Xinyi Zhou, Jin Huang, Dianqi Zhang, Zhenyu Qian, Xin Zuo, Yaoxiang Sun","doi":"10.1186/s13287-025-04330-5","DOIUrl":"https://doi.org/10.1186/s13287-025-04330-5","url":null,"abstract":"<p><p>Small extracellular vesicles (sEVs) are membrane-bound vesicles with a size of less than 200 nm, released by cells. Due to their relatively small molecular weight and ability to participate in intercellular communication, sEVs can serve not only as carriers of biomarkers for disease diagnosis but also as effective drug delivery agents. Furthermore, these vesicles are involved in regulating the onset and progression of various diseases, reflecting the physiological and functional states of cells. This paper introduces the classification of extracellular vesicles, with a focus on the extraction and identification of sEVs and their significant role in repair, diagnosis, and intercellular communication. Additionally, the paper addresses the engineering modification of sEVs to provide a reference for enhanced understanding and application.</p>","PeriodicalId":21876,"journal":{"name":"Stem Cell Research & Therapy","volume":"16 1","pages":"184"},"PeriodicalIF":7.1,"publicationDate":"2025-04-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12004682/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144042511","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Fine-tuning licensing strategies to boost MSC-based immunomodulatory secretome.","authors":"Maria Rossello-Gelabert, Manoli Igartua, Edorta Santos-Vizcaino, Rosa Maria Hernandez","doi":"10.1186/s13287-025-04315-4","DOIUrl":"https://doi.org/10.1186/s13287-025-04315-4","url":null,"abstract":"<p><strong>Background: </strong>Immune-mediated inflammatory diseases (IMIDs) are a major global health challenge, affecting millions of people and often lacking effective treatments. The mesenchymal stromal cell (MSC)-derived secretome has emerged as a promising therapeutic approach owing to its potent immunomodulatory properties. However, progress has been hindered by the lack of standardized protocols for inducing a robust immunomodulatory MSC phenotype.</p><p><strong>Methods: </strong>In this study, we focused on optimizing the MSC-derived secretome to enhance its ability to suppress activated immune cells. Specifically, we examined (1) the effects of IFN-γ and TNF-α, individually and in combination, to uncover potential synergy; (2) the ideal cytokine ratio and (3) concentration; (4) the best production time for the secretome; and (5) the impact of cellular confluence. These factors were systematically evaluated to assess their influence on cell behavior, viability, cytosolic content release, and the secretion of key immunomodulatory and regenerative factors.</p><p><strong>Results: </strong>Our results demonstrate that overnight licensing with a 1:1 ratio of IFN-γ and TNF-α at 60 ng/mL, followed by 48 h of incubation at 90% confluence, yields an optimized conditioned media (CM) with significantly enhanced immunomodulatory properties. Functional assays showed that this CM can inhibit human peripheral blood mononuclear cell (PBMC) activation with more than twice the effectiveness of suboptimal protocols. Additionally, we found that direct cell-cell contact was critical for inducing regulatory T cells (Tregs), highlighting the complex dynamics of immune regulation.</p><p><strong>Conclusions: </strong>These findings establish a robust and standardized MSC licensing protocol, paving the way for the development of innovative and effective therapies to combat IMIDs.</p><p><strong>Clinical trial number: </strong>Not applicable.</p>","PeriodicalId":21876,"journal":{"name":"Stem Cell Research & Therapy","volume":"16 1","pages":"183"},"PeriodicalIF":7.1,"publicationDate":"2025-04-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12004826/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144027183","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"KLF6-mediated glutamine metabolism governs odontogenic differentiation and matrix mineralization of dental pulp stem cells.","authors":"Wenzhi Wu, Zekai Xu, Yulian Zhang, Xiatong Zhang, Xiaoyuan Huang, Zhijian Xie, Zhuo Chen","doi":"10.1186/s13287-025-04308-3","DOIUrl":"https://doi.org/10.1186/s13287-025-04308-3","url":null,"abstract":"<p><strong>Background: </strong>When a tooth suffers severe injuries, dental pulp stem cells migrate and differentiate into odontoblast-like cells to form restorative dentin. Kruppel-like factor 6 (KLF6) activates the odontoblast differentiation of dental papilla cells during tooth development. However, the mechanisms by which KLF6 regulates the function of odontoblast-like cells differentiated from human dental pulp stem cells (hDPSCs) remain unknown.</p><p><strong>Methods: </strong>KLF6 was over-expressed or silenced by lentivirus transfection. Transcriptome sequencing and metabolomics were performed to reveal main changes in KLF6 high expressed hDPSCs. Mitochondrial morphology was observed by confocal microscope and cryo-transmission electron microscopy. Metabolic assays and metabolic flux were used to determine changes in cellular metabolic characteristics. Glutamine, glutamate, α-KG, and citrate concentrations were detected in cultured cells. Citrate and Ca concentration were detected in ECM. Adeno-associated virus were used to silence KLF6 in mice. A mouse dental injury model was established to investigate the role of KLF6 and glutamine metabolism in dentin repair in vivo.</p><p><strong>Results: </strong>RNA sequencing and metabolomics showed a remarkable influence on glutamine metabolism, mitochondrial respiration, and the TCA cycle by KLF6 overexpression. Metabolic assays and mitochondrial morphology observation found KLF6 promoted glutamine metabolism and mitochondrial function, and glutamine metabolism and mitochondrial respiration are enhanced during odontogenic differentiation of hDPSCs. Deprivation of glutamine inhibited mineralization of hDPSCs and restrained deposition of citrate and Ca in ECM. Increased glutamine entry into the tricarboxylic acid (TCA) cycle was both observed in differentiated hDPSCs and KLF6 overexpressed hDPSCs. ChIP-qPCR experiments revealed that KLF6 can directly bind to the promoter sequences of GLS1 and GDH. Supplementation of α-KG rescued suppression of odontogenic differentiation and mineralization induced by KLF6 knockdown. Inhibition of glutamine metabolism and knockdown of KLF6 attenuated tertiary dentin formation in vivo.</p><p><strong>Conclusions: </strong>Our study shows that KLF6 mediates biomineralization in the newly generated functional odontoblast-like cells differentiated from hDPSCs by altering cell metabolism preferences. KLF6 facilitated glutamine influx into the TCA cycle, leading to increased deposition of citrate in the ECM.These findings may inspire the development of novel strategies for reparative dentin formation.</p>","PeriodicalId":21876,"journal":{"name":"Stem Cell Research & Therapy","volume":"16 1","pages":"179"},"PeriodicalIF":7.1,"publicationDate":"2025-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12001474/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144049707","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}