Ultrasound-Mediated Lysozyme Microbubbles Targeting NOX4 Knockdown Alleviate Cisplatin-Exposed Cochlear Hair Cell...

Area of Science:

• Biomedical Engineering
• Molecular Biology
• Ototoxicity Research

Background:

• Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase 4 (NOX4) is crucial for reactive oxygen species (ROS) generation in cisplatin (CDDP)-induced ototoxicity.
• Current methods for targeted gene silencing face challenges in efficacy and delivery.

Purpose of the Study:

• To evaluate ultrasound-mediated lysozyme microbubble (USMB) cavitation for enhancing NOX4 siRNA transfection.
• To assess the efficacy of this method in reducing CDDP-induced ROS and cytotoxicity in vitro and ex vivo.

Main Methods:

• Lysozyme-shelled microbubbles (LyzMBs) were loaded with NOX4 siRNA (siNOX4/LyzMBs).
• siNOX4 delivery was compared using naked siRNA, LyzMB-mixed siRNA, and siNOX4-loaded LyzMBs with and without ultrasound (US) sonication.
• Transfection efficiency was measured by quantifying siRNA uptake in HEI-OC1 cells and mouse organ of Corti explants.

Main Results:

• USMB-mediated delivery of siNOX4 achieved high transfection efficacy (48.18%) in HEI-OC1 cells, significantly reducing CDDP-induced ROS.
• Ex vivo studies on mouse organ of Corti explants demonstrated a more potent NOX4 gene silencing effect with USMB and US sonication.
• siNOX4 loading onto LyzMBs stabilized the siRNA, enhancing transfection and silencing effects.

Conclusions:

• Ultrasound-mediated lysozyme microbubbles offer an effective strategy for NOX4 siRNA delivery.
• This USMB-based approach shows significant potential for mitigating CDDP-induced ototoxicity.
• The method holds promise for future clinical applications in preventing chemotherapy-related hearing damage.