Comparing transfection efficiency and safety for antisense oligodeoxyribonucleotide between phospholipids-based...

Area of Science:

• Biomedical Engineering
• Nanotechnology
• Molecular Biology

Background:

• Antisense oligodeoxynucleotides (AS-ODNs) are crucial for gene silencing.
• Phospholipids-based vectors are explored for drug/gene delivery.
• Ultrasound (US) can enhance cellular uptake.

Purpose of the Study:

• To compare the transfection efficiency and safety of AS-ODNs using two types of phospholipids-based vectors.
• To evaluate the impact of microbubbles and liposomes on AS-ODNs delivery.
• To assess the effect of ultrasound parameters on transfection outcomes.

Main Methods:

• Utilized AS-ODNs (HA824) with a luciferase reporter plasmid in SK-BR-3 breast cancer cells.
• Employed low-intensity ultrasound (US) with varying concentrations of microbubbles and liposomes.
• Assessed transfection efficiency via fluorescence microscopy, cell viability with propidium iodide assay, and gene expression changes using RT-PCR.
• Investigated membrane pore size alterations with Atomic Force Microscopy (AFM).

Main Results:

• Transfection efficiency increased with microbubble concentration but not liposome concentration.
• Optimal transfection efficiency with minimal cell viability impact was observed at 2% microbubble concentration.
• Excessive sonoporation led to significant membrane pore enlargement and reduced cell viability.

Conclusions:

• US-mediated AS-ODNs transfection is effectively enhanced by phospholipids-based microbubbles.
• This approach demonstrates a promising, safe, and efficient strategy for targeted gene silencing.
• Optimizing microbubble concentration is key to balancing transfection efficacy and cell safety.