Ultrasound and microbubble mediated plasmid DNA uptake: A fast, global and multi-mechanisms involved process

Area of Science:

• Biomedical Engineering
• Molecular Biology
• Cell Biology

Background:

• Ultrasound and microbubbles show promise for intracellular gene delivery.
• The precise mechanisms of plasmid DNA uptake via ultrasound and microbubbles remain unclear.

Purpose of the Study:

• To elucidate the detailed intracellular uptake process of plasmid DNA mediated by ultrasound and microbubbles.
• To highlight the specific role of microbubbles in this gene delivery method.

Main Methods:

• Utilized targeted microbubbles to localize ultrasound's acoustic force on cell membranes.
• Employed high-speed video microscopy and 3D confocal fluorescence microscopy.
• Investigated two ultrasound modes: high-pressure short pulse and low-pressure long pulse.

Main Results:

• High-pressure ultrasound induced reversible cell membrane disruption, allowing plasmid DNA entry.
• Plasmid DNA internalization and nuclear localization occurred rapidly (<2 min) under both ultrasound modes.
• Uptake was global, not restricted to microbubble attachment sites, indicating multi-mechanistic involvement.

Conclusions:

• Ultrasound-activated microbubbles enable fast and global plasmid DNA delivery into cells.
• The process involves multiple uptake pathways, including membrane disruption and other mechanisms.
• This technique offers a versatile approach for intracellular gene delivery.