Journal of extracellular biology最新文献

{"title":"Gut Microbiota and Bacterial Extracellular Vesicles: Emerging Roles in Myocardial Remodelling and Cardiac Health","authors":"Mingyang Chang,&nbsp;Tiantian Xia,&nbsp;Nan Zhang,&nbsp;Qianqian Zhao,&nbsp;Pan Shen,&nbsp;Ningning Wang,&nbsp;Chaoji Huangfu,&nbsp;Zhijie Bai,&nbsp;Dezhi Sun,&nbsp;Yangyi Hu,&nbsp;Shuman Li,&nbsp;Zhexin Ni,&nbsp;Wei Zhou,&nbsp;Yue Gao","doi":"10.1002/jex2.70079","DOIUrl":"10.1002/jex2.70079","url":null,"abstract":"<p>The gut microbiota, a collection of microorganisms residing within the human gastrointestinal tract, exerts profound effects on the health of the host. In recent years, studies have revealed that the gut microbiota influences not only the function of the digestive system but also has close associations with various systemic diseases, including cardiovascular diseases. Myocardial remodelling refers to the structural, functional and molecular changes in the myocardium that occur in response to alterations in load. This process encompasses changes such as myocardial hypertrophy, apoptosis, necrosis and myocardial fibrosis. Bacterial extracellular vesicles (BEVs) are small vesicles secreted by the gut microbiota that can carry bioactive substances such as proteins, lipids and nucleic acids, participating in intercellular communication. BEVs are capable of traversing the gut barrier and entering the bloodstream, thereby influencing the functional status of distant organs, including the heart. Under the condition of Myocardial remodelling, these BEVs may exert protective or detrimental effects on cardiomyocytes by modulating pathways such as inflammation, oxidative stress and apoptosis.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"4 8","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-08-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://isevjournals.onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.70079","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144814951","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cerebrospinal Fluid-Derived Extracellular Vesicles: A Proteomic and Transcriptomic Comparative Analysis of Enrichment Protocols","authors":"Marta García-Arauzo,&nbsp;Sandrine Reymond,&nbsp;Lyssia Gruaz,&nbsp;Domitille Schvartz,&nbsp;Natacha Civic,&nbsp;Mylène Docquier,&nbsp;Christine Deffert,&nbsp;Pascal Colosetti,&nbsp;Jean-Charles Sanchez,&nbsp;Claire Bridel","doi":"10.1002/jex2.70076","DOIUrl":"10.1002/jex2.70076","url":null,"abstract":"<p>Proteomic and transcriptomic analyses of cerebrospinal fluid (CSF)-derived extracellular vesicles (EVs) offer unique insights into molecular changes associated with central nervous system (CNS) diseases and may result in biomarker identification. No gold standard method to enrich EVs from CSF has been established, and head-to-head comparisons of outputs of different protocols are scarce. Using a large pool of CSF, we characterised the EV preparations resulting from four enrichment protocols and compared them in terms of yield and purity. We found that particles enriched by ultracentrifugation (UC) or a combination of ultrafiltration and size exclusion chromatography (UF-SEC) exhibited the typical morphological and biochemical characteristics of small EVs and were highly enriched in proteins and polyadenylated (polyA) transcripts associated with EV-related biological processes. UF-SEC preparations had higher particle yields, whilst more proteins were identified in UC preparations. Approximately 40% of the EV preparations’ proteome was not identified in unenriched CSF, among which a core proteome of 45 proteins was identified in 30 EV preparations from independent experiments, which may serve as CSF-derived EV markers. Enrichment scores to protein contaminants, albumin and apolipoprotein E were higher in UF-SEC preparations. In conclusion, all protocols analysed here resulted in enrichment of particles with small EV characteristics, with EV enrichments from UF-SEC resulting in the highest yield and purity.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"4 8","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-08-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://isevjournals.onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.70076","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144814663","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Profiling RNA Cargo in Extracellular Vesicles From hiPSC-Derived Neurons of Alzheimer's Disease Patients","authors":"Ram Sagar,&nbsp;Yiyao Huang,&nbsp;Daiyun Dong,&nbsp;Rachel J. Boyd,&nbsp;Waqar Ahmed,&nbsp;Kenneth W. Witwer,&nbsp;Vasiliki Mahairaki","doi":"10.1002/jex2.70074","DOIUrl":"10.1002/jex2.70074","url":null,"abstract":"<p>Alzheimer's disease (AD) is a major neurodegenerative disorder that affects more than 55 million people, with an incidence that is projected to triple by 2050. Despite continuous advancements in the field, reliable treatment and early detection strategies remain elusive. Extracellular vesicles (EVs) play a major role in cellular communication throughout the body. In this study, we assessed the cargo of neuronal-specific EVs for their potential as AD biomarkers. We isolated EVs released from iPSC-derived excitatory glutamatergic neurons generated from eight AD patients (ADiNEVs) and six healthy controls (iNEVs). We performed RNA-sequencing and identified significant differences in RNA cargo between ADiNEVs and iNEVs. Notably, fewer small nuclear RNAs (snRNAs) were found in ADiNEVs. RNA transcripts significantly more abundant in ADiNEVs included <i>MT-CO1, PRR32</i> and <i>IGSF8</i> messenger RNAs. We also observed fewer <i>XIST</i> long noncoding RNAs and <i>miR-7-5p</i> microRNA content in ADiNEVs. These findings suggest that precision medicine approaches, such as characterising the content of EVs from a patient's own cells, could advance early detection and management of AD.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"4 8","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-08-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.70074","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144782435","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Plasma Preparation Strategies for Extracellular Vesicle-Based Biomarkers in Metastatic Castration-Resistant Prostate Cancer","authors":"Prima Dewi Sinawang,&nbsp;Priyanka Multani,&nbsp;Mehmet O. Ozen,&nbsp;Jodie Wong,&nbsp;Demir Akin,&nbsp;Claire Hanson,&nbsp;Matt Larsen,&nbsp;Enos Ampaw,&nbsp;Matthew T. Rondina,&nbsp;Neal D. Tolley,&nbsp;Liang Wang,&nbsp;Brian T. Cunningham,&nbsp;Manish Kohli,&nbsp;Utkan Demirci","doi":"10.1002/jex2.70071","DOIUrl":"10.1002/jex2.70071","url":null,"abstract":"<p>Extracellular vesicles (EVs) offer a minimally invasive approach for cancer detection and monitoring. However, the lack of standardized methods for clinical biospecimen preparation and EV isolation limits the clinical utility of EV-based biomarker assessments. A targeted need exists for detailed analysis of plasma EV content. Our study investigates the impact of clinical sample preparation and our ExoTIC device on the quality of plasma-derived EVs and their RNA/protein cargo in metastatic castration-resistant prostate cancer (mCRPC) patients. We assessed sample preparation variables: blood anti-coagulant choice (EDTA or sodium citrate), type of plasma platelet fraction (platelet-rich or platelet-poor), and use of protease inhibitors. EVs were isolated via ExoTIC device, followed by EV characterization and biomarker analysis using nanoparticle tracking analysis (NTA), cryogenic electron microscopy, Western blot, and digital PCR (dPCR). We detected mCRPC-relevant proteins (ARv7 and PSMA) in EVs from all plasma sample types with different sample preparation variables. Additionally, our findings indicate that platelet-poor plasma (PPP) is optimal for detecting EV- and biologically associated mCRPC biomarker miR-375. In this pilot study (<i>n</i> = 3), elevated EV miR-375 levels in PPP samples from mCRPC patients experiencing disease progression during docetaxel treatment were associated with poor therapeutic response to docetaxel chemotherapy, which aligns with our preceding in vitro and in vivo study. Optimal biospecimen preparation for EV analysis could enhance detection accuracy and patient management, highlighting detection of plasma EV-associated mCRPC-specific marker proteins (ARv7 and PSMA) and microRNA miR-375.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"4 8","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.70071","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144740453","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Multiomics Profiling of Extracellular Vesicles Supports Their Involvement in Endothelial Senescence-Associated Vascular Dysfunction","authors":"Ryan E. Hogans,&nbsp;Yun Lin,&nbsp;Gabriela Grigorean,&nbsp;Ana Cristina Grodzki,&nbsp;Rachel R. Mizenko,&nbsp;Anne Knowlton,&nbsp;Randy P. Carney,&nbsp;Angie Gelli,&nbsp;Pamela J. Lein","doi":"10.1002/jex2.70078","DOIUrl":"10.1002/jex2.70078","url":null,"abstract":"<p>Dysfunction of vascular endothelium is characteristic of many aging-related diseases, including Alzheimer's disease (AD) and AD-related dementias (ADRD). Although it is widely posited that endothelial cell dysfunction contributes to the pathogenesis and/or progression of AD/ADRD, it is not clear how. A plausible hypothesis is that intercellular trafficking of extracellular vesicles (EVs) from senescent vascular endothelial cells promotes vascular endothelial cell dysfunction. To test this hypothesis, we compared the expression of proteins and miRNAs in EVs isolated from four sets of genetically identical early passage non-senescent (EP) versus late passage senescent (SEN) primary human coronary artery endothelial cells (HCAECs) derived from four donors. Proteomics and miRNA libraries constructed from these EV isolates were evaluated using FunRich gene ontology analysis to compare functional enrichment between EP and SEN endothelial cell EVs (ECEVs). Replicative senescence was associated with altered EV abundance and contents independent of changes in EV size. Unique sets of miRNAs and proteins were differentially expressed in SEN-ECEVs, including molecules related to cell adhesion, barrier integrity, receptor signalling, endothelial-mesenchymal transition and cell senescence. miR-181a-5p was the most upregulated miRNA in SEN-ECEVs, increasing &gt;5-fold. SEN-ECEV proteomes supported involvement in several pro-inflammatory pathways consistent with senescence and the senescence-associated secretory phenotype (SASP). These data indicate that SEN-ECEVs are enriched in bioactive molecules implicated in senescence-associated vascular dysfunction, blood–brain barrier impairment, and AD/ADRD pathology. These observations suggest involvement of SEN-ECEVs in the pathogenesis of vascular dysfunction associated with AD/ADRD.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"4 8","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-07-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.70078","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144740145","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Correction to Journal of Extracellular Biology Articles","authors":"","doi":"10.1002/jex2.70070","DOIUrl":"10.1002/jex2.70070","url":null,"abstract":"&lt;p&gt;Izhar, M. and Lesniak, M. S. (2025), Role of Extracellular Vesicles in the Pathogenesis of Brain Metastasis. J of Extracellular Bio., 4: e70051. https://doi.org/10.1002/jex2.70051&lt;/p&gt;&lt;p&gt;The data availability statement for this article was missing. The below data availability statement has been added to the article:&lt;/p&gt;&lt;p&gt;No new data were created or analyzed, and therefore, data sharing is not applicable.&lt;/p&gt;&lt;p&gt;Kim, K., Sohn, Y., &amp; Yeon, J. H. (2025). Anti-Ageing Activities of Nanovesicles Derived From &lt;i&gt;Artemisia&lt;/i&gt; princeps in Human Dermal Cells and Human Skin Model. &lt;i&gt;Journal of Extracellular Biology&lt;/i&gt;, 4, e70033. https://doi.org/10.1002/jex2.70033&lt;/p&gt;&lt;p&gt;The data availability statement for this article was missing. The below data availability statement has been added to the article:&lt;/p&gt;&lt;p&gt;The data that support the findings of this study are available on request from the corresponding author. The data are not publicly available due to privacy or ethical restrictions.&lt;/p&gt;&lt;p&gt;Qarawani, A., Naaman, E., Ben-Zvi Elimelech, R., Harel, M., Sigal-Dror, S., Ben-Zur, T., Ziv, T., Offen, D., &amp; Zayit-Soudry, S. (2025). Mesenchymal Stem Cell-Derived Exosomes Mitigate Amyloid β-Induced Retinal Toxicity: Insights From Rat Model and Cellular Studies. &lt;i&gt;Journal of Extracellular Biology&lt;/i&gt;, 4, e70024. https://doi.org/10.1002/jex2.70024&lt;/p&gt;&lt;p&gt;The data availability statement for this article was missing. The below data availability statement has been added to the article:&lt;/p&gt;&lt;p&gt;Data available on request from the authors.&lt;/p&gt;&lt;p&gt;Counil, H., Silva, R. O., Rabanel, J.-M., Zaouter, C., Haddad, M., Ben Khedher, M. R., Brambilla, D., Fülöp, T., Patten, S. A., &amp; Ramassamy, C. (2025). Brain Penetration of Peripheral Extracellular Vesicles From Alzheimer's Patients and Induction of Microglia Activation. &lt;i&gt;Journal of Extracellular Biology&lt;/i&gt;, 4, e70027. https://doi.org/10.1002/jex2.70027&lt;/p&gt;&lt;p&gt;The data availability statement for this article was missing. The below data availability statement has been added to the article:&lt;/p&gt;&lt;p&gt;The data that support the findings of this study are available on request from the corresponding author. The data are not publicly available due to privacy or ethical restrictions.&lt;/p&gt;&lt;p&gt;Mladenović, D., Brealey, J., Peacock, B., Koort, K., &amp; Zarovni, N. (2025). Quantitative Fluorescent Nanoparticle Tracking Analysis and Nano-Flow Cytometry Enable Advanced Characterization of Single Extracellular Vesicles. &lt;i&gt;Journal of Extracellular Biology&lt;/i&gt;, 4, e70031. https://doi.org/10.1002/jex2.70031&lt;/p&gt;&lt;p&gt;The data availability statement for this article was missing. The below data availability statement has been added to the article:&lt;/p&gt;&lt;p&gt;The data that supports the findings of this study are available in the supplementary material of this article.&lt;/p&gt;&lt;p&gt;Ghodsi, M., Cloos, A.-S., Lotens, A., De Bueger, M., Van Der Smissen, P., Henriet, P., Cellier, N., Pierreux, C. E., Najdovski, T., &amp; Tyteca, D. (2025). Development of an Easy Non","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"4 7","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-07-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.70070","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144705405","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Extracellular Vesicle Mitochondrial DNA Levels Are Associated With Chronic Kidney Disease and Mitochondrial Haplogroup in Obese Individuals","authors":"Jaida E. Morgan,&nbsp;Nicole Noren Hooten,&nbsp;Nicolle A. Mode,&nbsp;Ngozi Ezike,&nbsp;Alan B. Zonderman,&nbsp;Michele K. Evans","doi":"10.1002/jex2.70069","DOIUrl":"10.1002/jex2.70069","url":null,"abstract":"<p>Chronic kidney disease (CKD) and obesity are major chronic diseases in the United States. Although obesity is a risk factor for CKD, little is known about how obesity contributes to CKD. Due to their role as intercellular communicators, extracellular vesicles (EVs) may be a factor connecting obesity and CKD. Circulating cell-free mitochondrial DNA (ccf-mtDNA), a damage-associated molecular pattern molecule associated with inflammation, is associated with renal disease and may be encapsulated within EVs. In this longitudinal study, we isolated plasma EVs and analysed EV mtDNA levels in a cohort of African American and White obese middle-aged individuals who at visit 1 did not have CKD but developed CKD by visit 2 (<i>n</i> = 19; CKD group) and matched this group to controls who did not develop CKD by visit 2 (<i>n</i> = 56; control group). In our cross-sectional analyses at visit 1, we found significant interactions for EV mtDNA levels between race and CKD status, poverty status and CKD status, and sex and CKD status. EV mtDNA levels were significantly lower in participants within the African haplogroup who developed CKD compared to participants within the European haplogroup who developed CKD and the African haplogroup control group. In our longitudinal analyses using data from both visit 1 and visit 2, individuals who developed CKD had lower EV mtDNA levels. Stratification by haplogroup showed that among participants within the African haplogroup, those who developed CKD had significantly lower EV mtDNA levels than those in the control group. In conclusion, EV mtDNA levels were lower in individuals who develop CKD. Our findings demonstrate that CKD status and mtDNA haplogroup influence EV cargo in obese individuals.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"4 7","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-07-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.70069","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144672696","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Quantum Dot-Based Immunolabelling of Extracellular Vesicles and Detection Using Fluorescence-Based Nanoparticle Tracking Analysis","authors":"Eunyong Ha,&nbsp;Yewon Han,&nbsp;Minseop Kim,&nbsp;Zayakhuu Gerelkhuu,&nbsp;Sook Jin Kwon,&nbsp;Tae Hyun Yoon","doi":"10.1002/jex2.70072","DOIUrl":"10.1002/jex2.70072","url":null,"abstract":"<p>Extracellular vesicles (EVs) contain a variety of biomolecules, including DNA, RNA, lipids and proteins. They can interact with target cells to perform various functions, offering potential for therapeutic applications like drug delivery and diagnosis. The growing interest in EVs drives the need for robust methods for EV characterisation. One of the prevalent EV characterisation methods is scatter-based nanoparticle tracking analysis (Sc-NTA). This method measures the size and concentration of particles by tracking the scattered light from individual particles. However, Sc-NTA has limitations in selectivity, as it detects all scattered light and fails to distinguish EVs from other nanoparticles, such as protein aggregates. To overcome this limitation, fluorescence-based NTA (Fl-NTA) is being utilised, where fluorescence tagging is used to selectively detect EVs. In previous studies, lipophilic dyes were employed for membrane labelling, but this resulted in false-positive signals due to the staining of even non-vesicular extracellular particles (NVEPs). Immunolabelling methods using antibodies that specifically bind to EV-specific protein were also introduced; yet challenges with sensitivity and photostability of the organic dyes remained. To address the challenges, we conjugated quantum dots (QDs) to antibodies that specifically bind to EV-specific markers, CD9, CD63 and then immunolabelled the EVs. Labelling conditions were optimised to develop a robust protocol for QD-based immunolabelling. Detection sensitivity was evaluated by comparing QD-based immunolabelling with Alexa dye-based methods. Furthermore, size distribution analysis demonstrated the ability of QDs to detect smaller EV populations. Finally, subpopulations of EVs from various cell lines were profiled. This approach enhances the accurate characterisation of EVs, providing a reliable and reproducible method for EV quality control and improved insights into their heterogeneity.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"4 7","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-07-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.70072","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144673135","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cargo Analysis and MRI-Based Therapeutic Assessment of Iron Oxide Labelled Extracellular Vesicles of Hypoxia Human Stem Cells in Ischemic Stroke","authors":"Shannon Helsper,&nbsp;Li Sun,&nbsp;Richard Jeske,&nbsp;Chang Liu,&nbsp;Jacob Athey,&nbsp;Xuegang Yuan,&nbsp;Samuel C. Grant,&nbsp;Yan Li","doi":"10.1002/jex2.70063","DOIUrl":"10.1002/jex2.70063","url":null,"abstract":"<p>Human mesenchymal stem cells (hMSCs) have been under investigation in preclinical and clinical settings for treating neurological disorders in recent years. Predominantly due to paracrine effects <i>in vivo</i>, hMSC-secreted extracellular vesicles (EVs) are at the forefront of these investigations. In this study, the therapeutic efficacy of hypoxia hMSCs and the secreted EVs labelled with iron oxides was evaluated in a preclinical model of ischemic stroke. Transcriptome and proteomics analysis of hMSCs under hypoxia indicated alterations in metabolic pathways and EV biogenesis. Hypoxia preconditioning increased EV yield by 57% with similar EV size and exosomal marker expression. EV cargo analysis using proteomics and microRNA-sequencing revealed that hypoxia preconditioning upregulated expression of metabolic proteins related to hypoxia-inducible factor signalling, neurogenesis and EV biogenesis. Magnetic resonance imaging following <i>in vivo</i> administration of iron oxide-labelled hMSCs and EVs provided assessment of biodistribution and therapeutic efficacy. The results indicated differential recovery in sodium levels in rats following hMSC and EV administration compared to the vehicle-only group, supported by lactate levels and functional assessment. hMSC-EVs localized to the ischemic lesion and evoked a therapeutic response after a single bolus injection. This study has significance in developing human stem cell-free therapeutics for treating ischemic stroke.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"4 7","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.70063","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144647513","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"High-Sensitivity Detection of Urinary Extracellular Vesicles With Upconverting Nanoparticle-Based Lateral Flow Immunoassay","authors":"Md. Khirul Islam,&nbsp;Imran Mahmud,&nbsp;Klinton Ali,&nbsp;Teppo Salminen,&nbsp;Pekka Taimen,&nbsp;Peter J. Boström,&nbsp;Janne Leivo,&nbsp;Urpo Lamminmäki,&nbsp;Iida Martiskainen","doi":"10.1002/jex2.70053","DOIUrl":"10.1002/jex2.70053","url":null,"abstract":"<p>Urinary extracellular vesicles (uEVs) are well-known to express tetraspanin family membrane proteins abundantly on their surface. In this study, we aimed to develop an upconverting nanoparticle (UCNP)–based lateral flow immunoassay (UCNP-LFIA) designed for the rapid and high-sensitivity detection of CD63-positive uEVs for direct urinalysis. The assay utilizes UCNPs reporter to detect low concentrations of EVs. Minimally processed uEV samples from bladder cancer (BlCa) (<i>n</i> = 62), benign prostatic hyperplasia (BPH) (<i>n</i> = 50) and healthy (<i>n</i> = 30) individuals were tested in sandwich UCNP-LFIA format, capturing uEVs with the same anti-CD63 antibody conjugated to UCNP and immobilized on the test zone. After 80 min, the strips were read with an upconversion luminescence reader device. This UCNP-LFIA measured CD63-positive EVs with high sensitivity, exhibiting a limit of detection (LoD) of 4 × 10⁷ EVs/mL. The concentration of CD63-positive EVs in BlCa patients showed a 2.3-fold increase compared to benign conditions (<i>p</i> = 0.007), and a 16-fold increase compared to healthy controls (<i>p</i> = 0.00001). The results demonstrate the potential of UCNP-LFIA platform for sensitive and quantitative detection of uEVs, highlighting its promise as a tool for EV detection at point-of-care diagnostics.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"4 7","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-07-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.70053","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144573473","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}