Targeted Endothelial Gene Delivery by Ultrasonic Destruction of Magnetic Microbubbles Carrying Lentiviral Vectors
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Site specific vascular gene delivery is a promising tool for treatment of cardiovascular diseases. By combining ultrasound mediated microbubble destruction with site specific magnetic targeting of lentiviruses, we aimed to develop a technique suitable for systemic application.
The magnetic nanoparticle coupling to lipid microbubbles was confirmed by absorbance measurements. Association of fluorescent lentivirus to magnetic microbubbles (MMB) was determined by microscopy and flow cytometry. Functionality and efficiency of GFP-encoding lentiviral MMB transduction was evaluated by endothelial (HMEC) GFP expression and cytotoxicity was measured by MTT reduction.
Microbubbles with a mean diameter of 4.3 ± 0.04 μm were stable for 2 days, readily magnetizable and magnetically steerable in vitro and efficiently associated with lentivirus. Exposure of eGFP-encoding lentiviral MMB to human endothelial cells followed by application of an external static magnetic field (30 min) and ultrasonic destruction of the microbubbles did not markedly affect cellular viability. Finally, this combination led to a 30-fold increase in transduction efficiency compared to application of naked virus alone.
By associating microbubbles with magnetic iron nanoparticles, these function as carriers for lentiviruses achieving tissue specific deposition at the site of interest.
KEY WORDSendothelial cells gene delivery lentiviral transduction magnetic nanoparticles microbubbles ultrasound
ACKNOWLEDGMENTS & DISCLOSURES
This work was funded by the German Research Foundation (Deutsche Forschungsgemeinschaft, DFG) within the DFG Research Unit FOR917.
We thank Professor Kräusslich at the Department of Virology, University Clinic Heidelberg for kindly providing the pCHIV.eGFP lentiviral vector.
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