Optimization of ultrasound and microbubbles targeted gene delivery to cultured primary endothelial cells

Meijering, Bernadet D.M.; Henning, Robert H.; Gilst, Wiek H. van; Gavrilović, Ivana; Wamel, Annemieke van; Deelman, Leo E.

doi:10.1080/10611860701605088

Cited by 61 publications

(40 citation statements)

References 29 publications

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“…A key factor in gene delivery efficiency is the concentration of genetic material at the site of sonoporation (10)(11)(12)21). Although most published in vivo studies involved systemically administered mixtures of uncoupled plasmid DNA and MBs, efforts to improve ultrasound-mediated gene delivery efficiency have centered on functionalizing MBs to become carrier vehicles that directly bind and transport genetic cargo (14,15,22,23).…”

Section: Methodsmentioning

confidence: 99%

Cationic versus Neutral Microbubbles for Ultrasound-mediated Gene Delivery in Cancer

Wang¹,

Panje²,

Pysz³

et al. 2012

Radiology

103

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Purpose:To test whether plasmid-binding cationic microbubbles (MBs) enhance ultrasound-mediated gene delivery efficiency relative to control neutral MBs in cell culture and in vivo tumors in mice. Materials and Methods:Animal studies were approved by the institutional animal care committee. Cationic and neutral MBs were characterized in terms of size, charge, circulation time, and DNA binding. Click beetle luciferase (CBLuc) reporter plasmids were mixed with cationic or neutral MBs. The ability of cationic MBs to protect bound plasmids from nuclease degradation was tested by means of a deoxyribonuclease (DNase) protection assay. Relative efficiencies of ultrasound-mediated transfection (ultrasound parameters: 1 MHz, 1 W/cm 2 , 20% duty cycle, 1 minute) of CBLuc to endothelial cells by using cationic, neutral, or no MBs were compared in cell culture. Ultrasound-mediated gene delivery to mouse hind limb tumors was performed in vivo (n = 24) with insonation (1 MHz, 2 W/cm 2 , 50% duty cycle, 5 minutes) after intravenous administration of CBLuc with cationic, neutral, or no MBs. Tumor luciferase activity was assessed by means of serial in vivo bioluminescence imaging and ex vivo analysis. Results were compared by using analysis of variance. Results:Cationic

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Section: Methodsmentioning

confidence: 99%

Cationic versus Neutral Microbubbles for Ultrasound-mediated Gene Delivery in Cancer

Wang¹,

Panje²,

Pysz³

et al. 2012

Radiology

103

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“…Before UMTD, the Opticell chamber was mounted in the experimental acoustic setup, described in detail elsewhere 22 and, in brief, in the online data supplement, available at http://circres.ahajournals.org. Cells were exposed to sine-wave US-bursts with a 6.2% duty cycle and a 20-Hz pulse repetition frequency for 30 seconds.…”

Section: Ultrasound Exposurementioning

confidence: 99%

“…22 Cells between passage 3 and 7 were used for UMTD experiments. Forty-eight hours before UMTD, cells were seeded at 33% confluence to 1 of the 2 gas-permeable, US-transparent membranes of an Opticell cell culture chamber (Biocrystal, Westerville, Ohio).…”

Section: Cell Culturementioning

confidence: 99%

Ultrasound and Microbubble-Targeted Delivery of Macromolecules Is Regulated by Induction of Endocytosis and Pore Formation

Meijering

Juffermans

Wamel

et al. 2009

Circulation Research

Self Cite

412

372

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Abstract-Contrast microbubbles in combination with ultrasound (US) are promising vehicles for local drug and gene delivery. However, the exact mechanisms behind intracellular delivery of therapeutic compounds remain to be resolved. We hypothesized that endocytosis and pore formation are involved during US and microbubble targeted delivery (UMTD) of therapeutic compounds. Therefore, primary endothelial cells were subjected to UMTD of fluorescent dextrans (4.4 to 500 kDa) using 1 MHz pulsed US with 0.22-MPa peak-negative pressure, during 30 seconds. Fluorescence microscopy showed homogeneous distribution of 4.4-and 70-kDa dextrans through the cytosol, and localization of 155-and 500-kDa dextrans in distinct vesicles after UMTD. After ATP depletion, reduced uptake of 4.4-kDa dextran and no uptake of 500-kDa dextran was observed after UMTD. Independently inhibiting clathrin-and caveolae-mediated endocytosis, as well as macropinocytosis significantly decreased intracellular delivery of 4.4-to 500-kDa dextrans. Furthermore, 3D fluorescence microscopy demonstrated dextran vesicles (500 kDa) to colocalize with caveolin-1 and especially clathrin. Finally, after UMTD of dextran (500 kDa) into rat femoral artery endothelium in vivo, dextran molecules were again localized in vesicles that partially colocalized with caveolin-1 and clathrin. Together, these data indicated uptake of molecules via endocytosis after UMTD. In addition to triggering endocytosis, UMTD also evoked transient pore formation, as demonstrated by the influx of calcium ions and cellular release of preloaded dextrans after US and microbubble exposure. In conclusion, these data demonstrate that endocytosis is a key mechanism in UMTD besides transient pore formation, with the contribution of endocytosis being dependent on molecular size. Key Words: ultrasound microbubble targeted delivery Ⅲ cell membrane pore Ⅲ endocytosis Ⅲ dextran Ⅲ endothelial cells C onventional delivery methods for drugs or genes, such as systemic administration via intravenous injection or oral administration, often do not suffice for therapeutic compounds such as peptides, silencing RNAs and genes. 1 A recent development in delivery systems for therapeutic compounds is the microbubble-ultrasound (US) interaction. 2,3 Before its use as a clinical modality, it is of utmost importance to obtain new physiological insights into the mechanisms of uptake by US and microbubble-exposed cells.Microbubbles were originally developed as US contrast agents and are administered intravenously to the systemic circulation to enhance scattering of blood in echocardiography. They consist of a gas core stabilized with an encapsulation, ranging from 1 to 10 m in diameter. 4 Nowadays, research focuses on the use of US and microbubbles for therapeutic applications. It has been demonstrated that USexposed microbubbles can achieve safe and efficient local delivery of a variety of drugs 5,6 and genes.

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“…Plasmid DNA (42) and small interfering RNA (30) have been covalently bound via biotin-avidin-biotin linkages in pegylated lipoplexes attached to phospholipid-shelled bubbles (62). Several recent reviews have discussed a variety of bonding scenarios that may exist between microbubbles genes and/or small molecules (2,34,53,57). Today, a clear understanding of the interaction between microbubbles and plasmid DNA remains elusive and is ripe for investigation.…”

mentioning

confidence: 99%

“…A critical aspect of the ultrasound-directed, site-specific microbubble gene or drug delivery system is the formulation of the microbubble and therapeutic agent(s). For example, Optison (16,17,49), Definity (53), SonoVue (34), and liposome (57) delivery systems are (2). In other work, Sonazoid and small interfering RNA were added to a well containing cells and mixed gently just before exposure to ultrasound (39).…”

mentioning

confidence: 99%

Ultrasound-mediated targeted drug delivery: recent success and remaining challenges

Castle

Butts

Healey

et al. 2013

American Journal of Physiology-Heart and Circulatory Physiology

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contrast-enhanced ultrasound; gene therapy; microbubble drug delivery PRESENTLY, CONTRAST-ENHANCED ultrasound (CEUS) imaging focuses on diagnostic clinical applications. In the future, however, therapeutic uses of CEUS will create a paradigm shift for patient care and the pharmaceutical industry. Ultrasound contrast agents (UCAs) are composed of shelled microbubbles that serve as superior diagnostic agents while traversing the smallest of blood vessels resulting in unparalleled, real-time spatial and temporal imaging of intact tissues and organs. These microscopic, gas-filled microspheres acting as intravascular indicators represent ideal carrier vehicles for local delivery of ultrasound-directed drug and gene therapies. The conceptually simple application of external acoustic energy in the transformation of these inert, microspheres into powerful therapeutic systems has seemingly unlimited potential.Brief history of CEUS. All diagnostic imaging modalities use and require contrast effects to increase signal-to-noise ratios, permitting enhanced discrimination of the targets. Examples include the use of radiopaque contrast agents to create discrete image patterns using X-ray methods (ionizing radiation), thus creating enhanced detection of objects within the image plane. Similar to X-ray, positron emission tomography and radionuclide imaging procedures rely on radioactive emitters to highlight anatomy and provide information on cellular metabolism and physiology.

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Optimization of ultrasound and microbubbles targeted gene delivery to cultured primary endothelial cells

Cited by 61 publications

References 29 publications

Cationic versus Neutral Microbubbles for Ultrasound-mediated Gene Delivery in Cancer

Cationic versus Neutral Microbubbles for Ultrasound-mediated Gene Delivery in Cancer

Ultrasound and Microbubble-Targeted Delivery of Macromolecules Is Regulated by Induction of Endocytosis and Pore Formation

Ultrasound-mediated targeted drug delivery: recent success and remaining challenges

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