{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T09:09:00Z","timestamp":1789722540931,"version":"4.0.1"},"reference-count":36,"publisher":"MDPI AG","issue":"19","license":[{"start":{"date-parts":[[2024,10,7]],"date-time":"2024-10-07T00:00:00Z","timestamp":1728259200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004329","name":"Slovenian Research Agency","doi-asserted-by":"publisher","award":["P3-067"],"award-info":[{"award-number":["P3-067"]}],"id":[{"id":"10.13039\/501100004329","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004329","name":"Slovenian Research Agency","doi-asserted-by":"publisher","award":["20180033"],"award-info":[{"award-number":["20180033"]}],"id":[{"id":"10.13039\/501100004329","id-type":"DOI","asserted-by":"publisher"}]},{"name":"University Medical Centre Ljubljana","award":["P3-067"],"award-info":[{"award-number":["P3-067"]}]},{"name":"University Medical Centre Ljubljana","award":["20180033"],"award-info":[{"award-number":["20180033"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Diagnostics"],"abstract":"<jats:p>Background: High-grade serous carcinoma (HGSC) is the most lethal of gynecological cancers in developed countries. It usually presents late with non-specific symptoms and most cases are diagnosed at an advanced stage, with 5-year overall survival being around 40%. Biomarkers for screening and early diagnosis of this aggressive disease are, thus, a research priority. Extracellular vesicles (EVs) that reflect the cell of origin and that can be isolated from local fluid and plasma by minimally invasive liquid biopsy are such promising biomarkers. Besides EV concentration and molecular profile, which have been the main focus of research for many years, recent studies have also called attention to EV size distribution. The aim of our study was to evaluate the potential of EV concentration and size distribution in local fluid and plasma as diagnostic biomarkers for HGSC. Methods: Paired pretreatment ascites and plasma samples from 37 patients with advanced HGSC and paired pretreatment free peritoneal fluid (FPF) and plasma samples from 40 controls with benign ovarian pathology (BOP) were analyzed using nanoparticle tracking analysis (NTA). Results: We observed a significant difference in EV concentration in local fluid, but not in plasma, between HGSC patients and the control group. We also found a significant difference in EV size distribution in both local fluid and plasma between HGSC patients and the control group. The receiver operating characteristics (ROC) curve analysis of EV characteristics showed excellent diagnostic performance for the mode, D10, and D50 in local fluid and acceptable diagnostic performance for EV concentration and mean EV size in local fluid, as well as for the mode and D10 value in plasma. Conclusions: The results of our study show that EV concentration in local fluid and more importantly EV size distribution in both local fluid and plasma are significantly changed in the presence of HGSC. Future research of size-dependent molecular profiling of EVs could help identify novel diagnostic biomarkers for HGSC.<\/jats:p>","DOI":"10.3390\/diagnostics14192235","type":"journal-article","created":{"date-parts":[[2024,10,7]],"date-time":"2024-10-07T07:30:18Z","timestamp":1728286218000},"page":"2235","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Extracellular Vesicle Characteristics in Local Fluid and Plasma Measured by Nanoparticle Tracking Analysis Can Help Differentiate High-Grade Serous Carcinoma from Benign Ovarian Pathology"],"prefix":"10.3390","volume":"14","author":[{"given":"Maru\u0161a","family":"Herzog","sequence":"first","affiliation":[{"name":"Division of Gynecology and Obstetrics, University Medical Centre Ljubljana, SI-1000 Ljubljana, Slovenia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6249-5799","authenticated-orcid":false,"given":"Ivan","family":"Verdenik","sequence":"additional","affiliation":[{"name":"Division of Gynecology and Obstetrics, University Medical Centre Ljubljana, SI-1000 Ljubljana, Slovenia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katarina","family":"\u010cerne","sequence":"additional","affiliation":[{"name":"Institute of Pharmacology and Experimental Toxicology, Faculty of Medicine, University of Ljubljana, SI-1000 Ljubljana, Slovenia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7913-7699","authenticated-orcid":false,"given":"Borut","family":"Kobal","sequence":"additional","affiliation":[{"name":"Division of Gynecology and Obstetrics, University Medical Centre Ljubljana, SI-1000 Ljubljana, Slovenia"},{"name":"Department of Gynecology and Obstetrics, Faculty of Medicine, University of Ljubljana, SI-1000 Ljubljana, Slovenia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,10,7]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"672","DOI":"10.1093\/annonc\/mdz062","article-title":"ESMO\u2013ESGO Consensus Conference Recommendations on Ovarian Cancer: Pathology and Molecular Biology, Early and Advanced Stages, Borderline Tumours and Recurrent Disease","volume":"30","author":"Colombo","year":"2019","journal-title":"Ann. Oncol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"284","DOI":"10.3322\/caac.21456","article-title":"Ovarian Cancer Statistics, 2018","volume":"68","author":"Torre","year":"2018","journal-title":"CA Cancer J. Clin."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"4130","DOI":"10.7150\/thno.34692","article-title":"Liquid Biopsy in Ovarian Cancer: Recent Advances in Circulating Extracellular Vesicle Detection for Early Diagnosis and Monitoring Progression","volume":"9","author":"Chang","year":"2019","journal-title":"Theranostics"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Doyle, L., and Wang, M. (2019). Overview of Extracellular Vesicles, Their Origin, Composition, Purpose, and Methods for Exosome Isolation and Analysis. Cells, 8.","DOI":"10.3390\/cells8070727"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1535750","DOI":"10.1080\/20013078.2018.1535750","article-title":"Minimal Information for Studies of Extracellular Vesicles 2018 (MISEV2018): A Position Statement of the International Society for Extracellular Vesicles and Update of the MISEV2014 Guidelines","volume":"7","author":"Witwer","year":"2018","journal-title":"J. Extracell. Vesicles"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"e12404","DOI":"10.1002\/jev2.12404","article-title":"Minimal Information for Studies of Extracellular Vesicles (MISEV2023): From Basic to Advanced Approaches","volume":"13","author":"Welsh","year":"2024","journal-title":"J. Extracell. Vesicles"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Croft, P.K., Sharma, S., Godbole, N., Rice, G.E., and Salomon, C. (2021). Ovarian-Cancer-Associated Extracellular Vesicles: Microenvironmental Regulation and Potential Clinical Applications. Cells, 10.","DOI":"10.3390\/cells10092272"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"38","DOI":"10.1016\/j.ejcsup.2019.09.001","article-title":"The Influence of Secreted Factors and Extracellular Vesicles in Ovarian Cancer Metastasis","volume":"15","author":"Peinado","year":"2020","journal-title":"Eur. J. Cancer Suppl."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"395","DOI":"10.1586\/14789450.2016.1165613","article-title":"Extracellular Vesicles in Ovarian Cancer: Applications to Tumor Biology, Immunotherapy and Biomarker Discovery","volume":"13","author":"Nawaz","year":"2016","journal-title":"Expert Rev. Proteom."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Samuel, P., Mulcahy, L.A., Furlong, F., McCarthy, H.O., Brooks, S.A., Fabbri, M., Pink, R.C., and Carter, D.R.F. (2018). Cisplatin Induces the Release of Extracellular Vesicles from Ovarian Cancer Cells That Can Induce Invasiveness and Drug Resistance in Bystander Cells. Philos. Trans. R. Soc. B Biol. Sci., 373.","DOI":"10.1098\/rstb.2017.0065"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1226","DOI":"10.1016\/j.cell.2016.01.043","article-title":"Communication by Extracellular Vesicles: Where We Are and Where We Need to Go","volume":"164","author":"Tkach","year":"2016","journal-title":"Cell"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"490","DOI":"10.1038\/nbt.2886","article-title":"Label-Free Detection and Molecular Profiling of Exosomes with a Nano-Plasmonic Sensor","volume":"32","author":"Im","year":"2014","journal-title":"Nat. Biotechnol."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Zhang, W., Peng, P., Ou, X., Shen, K., and Wu, X. (2019). Ovarian Cancer Circulating Extracelluar Vesicles Promote Coagulation and Have a Potential in Diagnosis: An iTRAQ Based Proteomic Analysis. BMC Cancer, 19.","DOI":"10.1186\/s12885-019-6176-1"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"3033","DOI":"10.1039\/C6LC00279J","article-title":"Ultrasensitive Microfluidic Analysis of Circulating Exosomes Using a Nanostructured Graphene Oxide\/Polydopamine Coating","volume":"16","author":"Zhang","year":"2016","journal-title":"Lab. Chip"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1186\/s13048-018-0391-2","article-title":"Comparison of Benign Peritoneal Fluid- and Ovarian Cancer Ascites-Derived Extracellular Vesicle RNA Biomarkers","volume":"11","author":"Yamamoto","year":"2018","journal-title":"J. Ovarian Res."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Lucidi, A., Buca, D., Ronsini, C., Tinari, S., Bologna, G., Buca, D., Leombroni, M., Liberati, M., D\u2019Antonio, F., and Scambia, G. (2020). Role of Extracellular Vesicles in Epithelial Ovarian Cancer: A Systematic Review. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21228762"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Herzog, M., Verdenik, I., Kobal, B., and \u010cerne, K. (2024). Higher EpCAM-Positive Extracellular Vesicle Concentration in Ascites Is Associated with Shorter Progression-Free Survival of Patients with Advanced High-Grade Serous Carcinoma. Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms25126780"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"12391","DOI":"10.1002\/jev2.12391","article-title":"Correlation between Membrane Proteins and Sizes of Extracellular Vesicles and Particles: A Potential Signature for Cancer Diagnosis","volume":"12","author":"Zhai","year":"2023","journal-title":"J. Extracell. Vesicles"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"4141","DOI":"10.1007\/s00216-019-01616-5","article-title":"Size-Dependent Sub-Proteome Analysis of Urinary Exosomes","volume":"411","author":"Guan","year":"2019","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"9239","DOI":"10.1021\/acs.analchem.0c01572","article-title":"Deconstruction of Heterogeneity of Size-Dependent Exosome Subpopulations from Human Urine by Profiling N-Glycoproteomics and Phosphoproteomics Simultaneously","volume":"92","author":"Zheng","year":"2020","journal-title":"Anal. Chem."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Badovinac, D., Gori\u010dar, K., Lavrin, T., Zavrtanik, H., Dol\u017ean, V., Lenassi, M., and Toma\u017ei\u010d, A. (2023). Plasma Extracellular Vesicle Characteristics as Biomarkers of Resectability and Radicality of Surgical Resection in Pancreatic Cancer\u2014A Prospective Cohort Study. Cancers, 15.","DOI":"10.3390\/cancers15030605"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"\u010cerne, K., Kelhar, N., Resnik, N., Herzog, M., Vodnik, L., Verani\u010d, P., and Kobal, B. (2023). Characteristics of Extracellular Vesicles from a High-Grade Serous Ovarian Cancer Cell Line Derived from a Platinum-Resistant Patient as a Potential Tool for Aiding the Prediction of Responses to Chemotherapy. Pharmaceuticals, 16.","DOI":"10.20944\/preprints202305.0197.v1"},{"key":"ref_23","first-page":"5553","article-title":"Analysis of ESCRT Functions in Exosome Biogenesis, Composition and Secretion Highlights the Heterogeneity of Extracellular Vesicles","volume":"126","author":"Colombo","year":"2013","journal-title":"J. Cell Sci."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Yu, J., Sane, S., Kim, J.-E., Yun, S., Kim, H.-J., Jo, K.B., Wright, J.P., Khoshdoozmasouleh, N., Lee, K., and Oh, H.T. (2024). Biogenesis and Delivery of Extracellular Vesicles: Harnessing the Power of EVs for Diagnostics and Therapeutics. Front. Mol. Biosci., 10.","DOI":"10.3389\/fmolb.2023.1330400"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1182","DOI":"10.1111\/jth.12602","article-title":"Particle Size Distribution of Exosomes and Microvesicles Determined by Transmission Electron Microscopy, Flow Cytometry, Nanoparticle Tracking Analysis, and Resistive Pulse Sensing","volume":"12","author":"Coumans","year":"2014","journal-title":"J. Thromb. Haemost."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"44","DOI":"10.1016\/j.ab.2012.06.004","article-title":"Nanoparticle Analysis of Circulating Cell-Derived Vesicles in Ovarian Cancer Patients","volume":"428","author":"Atay","year":"2012","journal-title":"Anal. Biochem."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"26913","DOI":"10.3402\/jev.v3.26913","article-title":"Minimal Experimental Requirements for Definition of Extracellular Vesicles and Their Functions: A Position Statement from the International Society for Extracellular Vesicles","volume":"3","author":"Hill","year":"2014","journal-title":"J. Extracell. Vesicles"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"278","DOI":"10.1186\/1477-7819-12-278","article-title":"Control Values of Ovarian Cancer Tumor Markers and Standardisation of a Protocol for Sampling Peritoneal Fluid and Performing Washing during Laparoscopy","volume":"12","author":"Jerman","year":"2014","journal-title":"World J. Surg. Oncol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1632","DOI":"10.1161\/CIRCRESAHA.117.309417","article-title":"Methodological Guidelines to Study Extracellular Vesicles","volume":"120","author":"Coumans","year":"2017","journal-title":"Circ. Res."},{"key":"ref_30","first-page":"35","article-title":"The Effect of Equex STM in Freezing Media on Post Thaw Motility, Viability and Dna Integrity of Frozen\u2014Thawed Ram Spermatozoa","volume":"51","author":"Sterbenc","year":"2014","journal-title":"Slov. Vet. Res."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"107","DOI":"10.5414\/CNP96S19","article-title":"Optimization of Isolation Protocol and Characterization of Urinary Extracellular Vesicles as Biomarkers of Kidney Allograft Injury","volume":"96","author":"Sedej","year":"2021","journal-title":"Clin. Nephrol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"104","DOI":"10.1186\/s12967-021-02775-9","article-title":"Extracellular Vesicles Isolated by Size-Exclusion Chromatography Present Suitability for RNomics Analysis in Plasma","volume":"19","author":"Yang","year":"2021","journal-title":"J. Transl. Med."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Kurtjak, M., Kere\u00efche, S., Klepac, D., Kri\u017ean, H., Per\u010di\u0107, M., Kru\u0161i\u0107 Ali\u0107, V., Lavrin, T., Lenassi, M., Wechtersbach, K., and Kojc, N. (2022). Unveiling the Native Morphology of Extracellular Vesicles from Human Cerebrospinal Fluid by Atomic Force and Cryogenic Electron Microscopy. Biomedicines, 10.","DOI":"10.3390\/biomedicines10061251"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"4337","DOI":"10.3390\/cimb46050264","article-title":"Effect of the 35 Nm and 70 Nm Size Exclusion Chromatography (SEC) Column and Plasma Storage Time on Separated Extracellular Vesicles","volume":"46","author":"Balbisi","year":"2024","journal-title":"Curr. Issues Mol. Biol."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Englisz, A., Smycz-Kuba\u0144ska, M., and Mielczarek-Palacz, A. (2024). Sensitivity and Specificity of Selected Biomarkers and Their Combinations in the Diagnosis of Ovarian Cancer. Diagnostics, 14.","DOI":"10.3390\/diagnostics14090949"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"4213","DOI":"10.1007\/s13277-015-4105-8","article-title":"Characterization of Exosomes Derived from Ovarian Cancer Cells and Normal Ovarian Epithelial Cells by Nanoparticle Tracking Analysis","volume":"37","author":"Zhang","year":"2016","journal-title":"Tumor Biol."}],"container-title":["Diagnostics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2075-4418\/14\/19\/2235\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T16:12:09Z","timestamp":1760112729000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2075-4418\/14\/19\/2235"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,10,7]]},"references-count":36,"journal-issue":{"issue":"19","published-online":{"date-parts":[[2024,10]]}},"alternative-id":["diagnostics14192235"],"URL":"https:\/\/doi.org\/10.3390\/diagnostics14192235","relation":{},"ISSN":["2075-4418"],"issn-type":[{"value":"2075-4418","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,10,7]]}}}