Efficient dispersion of aggregated extracellular vesicles: a comparative study of water-bath sonication and regular pipetting

Abstract Extracellular vesicles (EVs) are nano-sized particles released by various cell types that facilitate intercellular communication by transferring bioactive molecules. Owing to their biocompatibility, EVs are widely used as drug delivery vehicles. However, freezing EVs at – 70 °C can induce a...

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Main Authors: Suryeon Kim, Inhwan Oh, Juhyun Kim, Nayeong Lee, Jun Go, Yang Jin, Won June Kim, Heedoo Lee
Format: Article
Language:English
Published: Nature Portfolio 2025-07-01
Series:Scientific Reports
Subjects:
Online Access:https://doi.org/10.1038/s41598-025-10050-w
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author Suryeon Kim
Inhwan Oh
Juhyun Kim
Nayeong Lee
Jun Go
Yang Jin
Won June Kim
Heedoo Lee
author_facet Suryeon Kim
Inhwan Oh
Juhyun Kim
Nayeong Lee
Jun Go
Yang Jin
Won June Kim
Heedoo Lee
author_sort Suryeon Kim
collection DOAJ
description Abstract Extracellular vesicles (EVs) are nano-sized particles released by various cell types that facilitate intercellular communication by transferring bioactive molecules. Owing to their biocompatibility, EVs are widely used as drug delivery vehicles. However, freezing EVs at – 70 °C can induce aggregation, reducing their effective concentration and drug delivery efficiency. In this study, we evaluated the use of water-bath sonication to disperse aggregated EVs and compared its effectiveness with regular pipetting. Frozen EVs showed reduced total concentration and increased aggregation relative to fresh EVs. Sonication at power level 3 (40 kHz, 100 W) significantly increased EV concentration and reduced aggregation. Unlike pipetting, only sonication effectively dispersed aggregated EVs, though subsequent pipetting caused reaggregation. In vivo, aggregated EVs were detected in the bronchoalveolar lavage fluid of mice treated with frozen EVs, whereas mice receiving sonicated EVs exhibited fewer aggregates and enhanced cellular uptake. Molecular dynamics simulations supported the effectiveness of sonication in dispersing EVs. In conclusion, water-bath sonication is a simple and effective method to restore the functionality of freeze-thawed EVs, improving their intracellular delivery and therapeutic potential.
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issn 2045-2322
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spelling doaj-art-6626ed8455fe4bbbb93c95c7fc15a9832025-08-20T04:02:45ZengNature PortfolioScientific Reports2045-23222025-07-0115111010.1038/s41598-025-10050-wEfficient dispersion of aggregated extracellular vesicles: a comparative study of water-bath sonication and regular pipettingSuryeon Kim0Inhwan Oh1Juhyun Kim2Nayeong Lee3Jun Go4Yang Jin5Won June Kim6Heedoo Lee7Department of Biology and Chemistry, Changwon National UniversityDepartment of Biology and Chemistry, Changwon National UniversityDepartment of Biology and Chemistry, Changwon National UniversityDepartment of Biology and Chemistry, Changwon National UniversityDepartment of Biology and Chemistry, Changwon National UniversityDivision of Pulmonary and Critical Care Medicine, Department of Medicine, Boston University School of MedicineDepartment of Biology and Chemistry, Changwon National UniversityDepartment of Biology and Chemistry, Changwon National UniversityAbstract Extracellular vesicles (EVs) are nano-sized particles released by various cell types that facilitate intercellular communication by transferring bioactive molecules. Owing to their biocompatibility, EVs are widely used as drug delivery vehicles. However, freezing EVs at – 70 °C can induce aggregation, reducing their effective concentration and drug delivery efficiency. In this study, we evaluated the use of water-bath sonication to disperse aggregated EVs and compared its effectiveness with regular pipetting. Frozen EVs showed reduced total concentration and increased aggregation relative to fresh EVs. Sonication at power level 3 (40 kHz, 100 W) significantly increased EV concentration and reduced aggregation. Unlike pipetting, only sonication effectively dispersed aggregated EVs, though subsequent pipetting caused reaggregation. In vivo, aggregated EVs were detected in the bronchoalveolar lavage fluid of mice treated with frozen EVs, whereas mice receiving sonicated EVs exhibited fewer aggregates and enhanced cellular uptake. Molecular dynamics simulations supported the effectiveness of sonication in dispersing EVs. In conclusion, water-bath sonication is a simple and effective method to restore the functionality of freeze-thawed EVs, improving their intracellular delivery and therapeutic potential.https://doi.org/10.1038/s41598-025-10050-wExtracellular vesiclesParticle aggregationWater-bath sonicationRegular pipetting
spellingShingle Suryeon Kim
Inhwan Oh
Juhyun Kim
Nayeong Lee
Jun Go
Yang Jin
Won June Kim
Heedoo Lee
Efficient dispersion of aggregated extracellular vesicles: a comparative study of water-bath sonication and regular pipetting
Scientific Reports
Extracellular vesicles
Particle aggregation
Water-bath sonication
Regular pipetting
title Efficient dispersion of aggregated extracellular vesicles: a comparative study of water-bath sonication and regular pipetting
title_full Efficient dispersion of aggregated extracellular vesicles: a comparative study of water-bath sonication and regular pipetting
title_fullStr Efficient dispersion of aggregated extracellular vesicles: a comparative study of water-bath sonication and regular pipetting
title_full_unstemmed Efficient dispersion of aggregated extracellular vesicles: a comparative study of water-bath sonication and regular pipetting
title_short Efficient dispersion of aggregated extracellular vesicles: a comparative study of water-bath sonication and regular pipetting
title_sort efficient dispersion of aggregated extracellular vesicles a comparative study of water bath sonication and regular pipetting
topic Extracellular vesicles
Particle aggregation
Water-bath sonication
Regular pipetting
url https://doi.org/10.1038/s41598-025-10050-w
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