Transient transmembrane release of green fluorescent proteins with sonoporation

IEEE Transactions on Ultrasonics, Ferroelectrics, and Frequency Control
Kadija KaddurA Bouakaz

Abstract

Microbubbles under ultrasound (US) activation are assumed to induce pore formation in the plasma membrane, causing its permeabilization and hence molecule incorporation from the extracellular environment. In this study, we investigated whether this permeabilization also engenders a transient release of small molecules from the cytosol of mammalian eukaryotic cells under the combined action of US and microbubbles. Using Hela cells stably expressing the enhanced green fluorescent protein (EGFP) gene, the release of EGFP was evaluated by flow cytometry in terms of the percentage of EGFP-positive cells (EGFP + cells) and the mean cell fluorescence intensity (MFI). Sonoporation was performed at 1 MHz, with peak negative pressures ranging from 0.2 to 0.6 MPa, duty cycles of 40% and 75% and a repetition rate of 10 kHz. The results showed that the insonation of Hela-EGFP cells at the peak negative pressure 400 kPa and the 75% duty cycle for 2 min in the presence of microbubbles induced a 60% decrease in both EGFP+ cells percentage and MFI. Our results demonstrate that the reduction of cell fluorescence is attributed to the EGFP release. Most importantly, this EGFP release was not due to lethal effects of sonoporation because the EGFP e...Continue Reading

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Citations

Nov 3, 2012·Therapeutic Delivery·Anthony DelalandeChantal Pichon
Nov 26, 2013·Advanced Drug Delivery Reviews·I LentackerC T W Moonen
Jun 1, 2014·Journal of Drug Targeting·Aya ZeghimiJean-Michel Escoffre
Sep 6, 2011·Journal of Controlled Release : Official Journal of the Controlled Release Society·Anthony DelalandeChantal Pichon
Jan 5, 2013·IEEE Transactions on Ultrasonics, Ferroelectrics, and Frequency Control·Jean-Michel EscoffreAyache Bouakaz
Apr 10, 2013·Gene·Anthony DelalandeChantal Pichon
Sep 18, 2016·Ultrasound in Medicine & Biology·Martynas MaciulevičiusSaulius Šatkauskas
May 20, 2017·BMC Biotechnology·Farah HusseinRaffi Karshafian
Nov 29, 2017·Bioscience Reports·Anthony DelalandeChantal Pichon
Feb 19, 2017·Molecular Imaging and Biology : MIB : the Official Publication of the Academy of Molecular Imaging·B H A LammertinkC Bos
Jan 18, 2019·Langmuir : the ACS Journal of Surfaces and Colloids·Silke RooversIne Lentacker

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