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Open AccessCommunication
Int. J. Mol. Sci. 2017, 18(8), 1610; doi:10.3390/ijms18081610

Microbubbles-Assisted Ultrasound Triggers the Release of Extracellular Vesicles

1
Imaging Division, University Medical Center Utrecht, Utrecht, 3584 CX, The Netherland
2
Department of Clinical Chemistry and Hematology, University Medical Center Utrecht, Utrecht, 3584 CX, The Netherlands
*
Author to whom correspondence should be addressed.
Received: 25 June 2017 / Revised: 12 July 2017 / Accepted: 16 July 2017 / Published: 25 July 2017
(This article belongs to the Section Molecular Pathology, Diagnostics, and Therapeutics)
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Abstract

Microbubbles-assisted ultrasound (USMB) has shown promise in improving local drug delivery. The formation of transient membrane pores and endocytosis are reported to be enhanced by USMB, and they contribute to cellular drug uptake. Exocytosis also seems to be linked to endocytosis upon USMB treatment. Based on this rationale, we investigated whether USMB triggers exocytosis resulting in the release of extracellular vesicles (EVs). USMB was performed on a monolayer of head-and-neck cancer cells (FaDu) with clinically approved microbubbles and commonly used ultrasound parameters. At 2, 4, and 24 h, cells and EV-containing conditioned media from USMB and control conditions (untreated cells, cells treated with microbubbles and ultrasound only) were harvested. EVs were measured using flow cytometric immuno-magnetic bead capture assay, immunogold electron microscopy, and western blotting. After USMB, levels of CD9 exposing-EVs significantly increased at 2 and 4 h, whereas levels of CD63 exposing-EVs increased at 2 h. At 24 h, EV levels were comparable to control levels. EVs released after USMB displayed a heterogeneous size distribution profile (30–1200 nm). Typical EV markers CD9, CD63, and alix were enriched in EVs released from USMB-treated FaDu cells. In conclusion, USMB treatment triggers exocytosis leading to the release of EVs from FaDu cells. View Full-Text
Keywords: sonoporation; exocytosis; endocytosis; exosomes; microvesicles; drug delivery sonoporation; exocytosis; endocytosis; exosomes; microvesicles; drug delivery
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Yuana, Y.; Jiang, L.; Lammertink, B.H.A.; Vader, P.; Deckers, R.; Bos, C.; Schiffelers, R.M.; Moonen, C.T. Microbubbles-Assisted Ultrasound Triggers the Release of Extracellular Vesicles. Int. J. Mol. Sci. 2017, 18, 1610.

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