Search results for "Sonoporation"
showing 10 items of 10 documents
Physical Methods of Gene Delivery
2017
Gene therapy can be defined as the use of nucleic acids (NAs) as medicines with the aim of correcting a deficient gene expression, introducing new functions in the cell, repairing mutations and modulating the gene expression. Two main classes of vectors, viral and nonviral, have been used for gene delivery in order to avoid the NAs hydrolysis by tissue nucleases and improve their cellular uptake. The ideal gene delivery vector should offer high transfection efficacy, cell specificity and low toxicity. However, the immunogenic and mutagenic side effects of viral vector as well as toxicity and low efficacy of nonviral carriers are limiting their application. In this respect, naked NAs deliver…
Noninvasive optical diagnostics of enhanced green fluorescent protein expression in skeletal muscle for comparison of electroporation and sonoporatio…
2015
We highlight the options available for noninvasive optical diagnostics of reporter gene expression in mouse tibialis cranialis muscle. An in vivo multispectral imaging technique combined with fluorescence spectroscopy point measurements has been used for the transcutaneous detection of enhanced green fluorescent protein (EGFP) expression, providing information on location and duration of EGFP expression and allowing quantification of EGFP expression levels. For EGFP coding plasmid (pEGFP-Nuc Vector, 10 μg/50 ml 10 μg/50 ml ) transfection, we used electroporation or ultrasound enhanced microbubble cavitation [sonoporation (SP)]. The transcutaneous EGFP fluorescence in live mice was monit…
FRET-based method for evaluation of the efficiency of reversible and irreversible sonoporation.
2017
It is widely known that not all of the treated cells survive after introduction of exogenous molecules via any physical method. Therefore, it is important to develop methods that would allow simultaneous evaluation of both molecular delivery efficiency and cell viability. This study presents Forster resonance energy transfer (FRET)-based method that allows molecular transfer and cell viability evaluation in a single measurement by employing two common fluorescent dyes, namely, ethidium bromide and trypan blue. The method has been validated using cell sonoporation. The FRET-based method allows the efficiency evaluation of both reversible and irreversible sonoporation in a single experiment. …
Fluorescence spectroscopy for estimation of anticancer drug sonodestruction in vitro
2013
The effect of ultrasound exposure on bleomycin fluorescence and pharmacological properties is studied. Bleomycin was treated by ultrasound for 7 min. Bleomycin fluorescence was measured during ultrasound exposure by means of fiber-optic spectrometry. Cell colony test was used to evaluate blemycin cytotoxity before and after ultrasound exposure.
A Setup for Microscopic Studies of Ultrasounds Effects on Microliters Scale Samples: Analytical, Numerical and Experimental Characterization
2021
International audience; Sonoporation is the process of cell membrane permeabilization, due to exposure to ultrasounds. There is a lack of consensus concerning the mechanisms of sonoporation: Understanding the mechanisms of sonoporation refines the choice of the ultrasonic parameters to be applied on the cells. Cells’ classical exposure systems to ultrasounds have several drawbacks, like the immersion of the cells in large volumes of liquid, the nonhomogeneous acoustic pressure in the large sample, and thus, the necessity for magnetic stirring to somehow homogenize the exposure of the cells. This article reports the development and characterization of a novel system allowing the exposure to …
Assessment of efficiencies of electroporation and sonoporation methods by using fluorescence RGB imaging method
2013
Simple RGB method for fluorescence in vivo imaging is presented to assess efficiency of electroporation and sonoporation methods by measuring distribution and accumulation of green fluorescence protein (GFP) concentration. 20 laboratory measurements were performed on mice to test the method.
Sonoporation, a redefined ultrasound modality as therapeutic aid: a review.
2011
Traditionally a diagnostic modality, ultrasound is emerging as a promising tool for non-invasive therapy, drug delivery, and gene therapy. The ultrasound is a mechanical wave energy generated in a medium as oscillating pressure in space and time at frequencies above 20 kHz, beyond the audible range. The ultrasound exposure generates bioeffects resulting in tissue heating, shear stress, and cavitation, which have been exploited for therapeutic applications. Ultrasound cavitation, enhanced by injected micro bubbles, perturbs cell membrane structures to cause sonoporation and increases the permeability to bioactive materials. Ultrasound-mediated gene delivery has been applied to heart, blood v…
pEGFP transfection into murine skeletal muscle by electrosonoporation
2017
In this study, we aimed to determine whether the combination of electroporation (EP) and ultrasound (US) waves (sonoporation) can affect the plasmid DNA transfection to mice tibialis cranialis muscle. Multispectral imaging technique combined with fluorescence spectroscopy point measurements has been used for the transcutaneous detection of enhanced green fluorescent protein (EGFP) fluorescence, providing information on location and duration of EGFP expression. We found that electrosonoporation, commonly enhancing pDNA transfection in vitro , had no positive effect on EGFP transfection efficiency increase in vivo with respect to electroporation alone. We presume that this may be associated w…
Application of fluorescence spectroscopy and multispectral imaging for non-invasive estimation of GFP transfection efficiency
2014
Electroporation and ultrasound induced sonoporation has been showed to induce plasmid DNA transfection to the mice tibialis cranialis muscle. It offers new prospects for gene therapy and cancer treatment. However, numerous experimental data are still needed to deliver the plausible explanation of the mechanisms governing DNA electro- or sono-transfection, as well as to provide the updates on transfection protocols for transfection efficiency increase. In this study we aimed to apply non-invasive optical diagnostic methods for the real time evaluation of GFP transfection levels at the reduced costs for experimental apparatus and animal consumption. Our experimental set-up allowed monitoring …
A versatile ultrasound system for in vitro experiments
2015
Objective One of the most difficult tasks to achieve with the available instrumentations used to study the interaction between ultrasound (US) and cellular model systems is to design an experiment, where only the effects of one physical parameter at a time is evaluated, while all the others are kept constant. The set-ups are usually custom-made, often by means of clinical instrument intended for a different therapeutic purpose. Furthermore, the results are not strictly comparable with others obtained with techniques considered standard in molecular and cellular biology at this time, because there is the need to use non-standard devices to contain biological samples. Sterility, as well as te…