Microbubbles-Assisted Ultrasound Triggers the Release of Extracellular Vesicles.

Authors:
Yuana Yuana
Yuana Yuana
Academic Medical Centre of the University of Amsterdam
Netherlands
Linglei Jiang
Linglei Jiang
Pharmaceutical Analytical and Solid-State Chemistry Research Center
Pieter Vader
Pieter Vader
University Medical Center Utrecht
Netherlands
Roel Deckers
Roel Deckers
University Medical Center Utrecht
Netherlands
Clemens Bos
Clemens Bos
University Medical Center Utrecht
Netherlands
Raymond M Schiffelers
Raymond M Schiffelers
Utrecht University
Netherlands
Chrit T Moonen
Chrit T Moonen
University Medical Center Utrecht
Netherlands

Int J Mol Sci 2017 Jul 25;18(8). Epub 2017 Jul 25.

Imaging Division, University Medical Center Utrecht, Utrecht, 3584 CX, The Netherland.

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.

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http://dx.doi.org/10.3390/ijms18081610DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5578002PMC

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July 2017
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