Development and Characterization of Ultrasound Activated Lipopolyplexes for Enhanced Transfection by Low Frequency Ultrasound in In Vitro Tumor Model

Abstract

This work focuses on the development of ultrasound contrast vesicles for ultrasound‐mediated enhanced transfection of nucleic acids in the cancer cells and projects its application as a tool for diagnostic imaging. The ultrasound contrast vesicles are stable, anionic, nanoscaled vesicles with ultrasound contrast equivalent to the commercially available SonoVue. These anionic lipid vesicles establish electrostatic interaction with cationic polyplexes based on linear polyethylenimine (22kDa) forming lipopolyplexes with ultrasound contrast. The lipopolyplexes are characterized regarding shape, size, and zeta potential. When exposed to low frequency ultrasound, these carriers show elevated transfection efficiency and reduced cytotoxicity. The effect of post‐transfection ultrasound on cellular uptake of lipopolyplexes is also evaluated. An analogous transfection is also observed in the tumor mimicking multicellular 3D spheroid culture of ovarian cancer cells. The emergence of tumor imaging and enhanced gene delivery by medical ultrasound, a noninvasive imaging modality, is considered paving the way for efficient theranostic gene therapy.

Lipodendriplexes mediated enhanced gene delivery: a cellular to pre-clinical investigation

“…From our experiment, it was observed that the lipid covering of dendriplexes enhanced the penetration and gene expression of the nanocarrier system in the spheroidal matrix system and demonstrated a superior GFP expression while a lower transfection efficiency was observed with the naked dendriplexes system (Fig. 5a) 45 . The results obtained with multicellular spheroid coincide with that observed in the 2D culture of HEK-293 cells.…”

Non‐Viral Nucleic Acid Delivery System for RNA Therapeutics

“…Ultrasound was first used in the mid‐1990s to mediate the in vitro transfection of DNA. [ 237 ] It was not until 2000 that ultrasound was used in vivo to transfer nucleic acid molecules to muscle, [ 238 ] solid tumor, [ 239 ] liver, [ 240 ] kidney, [ 241 ] and heart. [ 242 ] This method can realize the gene transfection of visceral organs [ 243 ] without surgery, and the use of ultrasound contrast agent or microbubbles filled with compressed air can further improve the gene expression level.…”

Polyethyleneimine-Based Lipopolyplexes as Carriers in Anticancer Gene Therapies

“…Another unusual approach focuses on improving the lipopolyplex cellular uptake via local use of low-frequency ultrasound (3 MHz) [ 121 ]. In in vitro studies such a procedure allows better penetration of the proliferating (outermost) layers of 3D spheroid culture of ovarian cancer SKOV-3 cells by vesicles carrying both an ultrasound contrasting agent (inside the carriers) and PEI-based polyplexes (present on the surface), even in the presence of endocytosis inhibitors.…”