Microfluidic-Driven Lipid Nanoparticles for Improved miRNA Delivery via Endo-Lysosomal Trafficking Optimization

Area of Science:

• Biotechnology
• Nanomedicine
• Gene Delivery

Background:

• Lipid nanoparticles (LNPs) are crucial for delivering small interfering RNA (siRNA) and microRNA (miRNA).
• Optimizing LNP physicochemical properties is essential for effective gene silencing.
• Understanding the impact of post-processing on LNP characteristics is vital for reproducible and efficient miRNA delivery systems.

Purpose of the Study:

• To investigate how various post-processing techniques influence the characteristics of miRNA-loaded lipid nanoparticles (LNP-miRNA).
• To correlate LNP physicochemical properties with their in vitro transfection performance and intracellular miRNA delivery.
• To establish a framework for optimizing non-viral miRNA delivery systems through tailored post-processing.

Main Methods:

• Comparison of blank and miRNA-loaded LNPs (LNP-miRNA) regarding size, polydispersity index, zeta potential, electrophoretic mobility, and conductivity.
• Application of post-processing techniques: sonication, filtration, dialysis, and thermal treatment.
• In vitro transfection models to assess intracellular trafficking and miRNA persistence.

Main Results:

• miRNA encapsulation increased LNP size by 43.6% due to structural changes.
• Sonication and filtration reduced particle size and improved uniformity, enhancing colloidal stability.
• Dialysis refined particle size but decreased electrophoretic mobility; however, dialyzed LNPs showed earlier intracellular availability and prolonged miRNA persistence.
• Sonication and filtration increased zeta potential to +29.3 mV, enhancing colloidal stability, while dialysis reduced it to +1.9 mV.

Conclusions:

• Post-processing methods significantly alter LNP characteristics, impacting both stability and transfection efficiency.
• While sonication and filtration yield favorable physicochemical properties for colloidal stability, dialysis enhances intracellular miRNA delivery and persistence.
• Tailoring post-processing techniques is key to optimizing LNP-based miRNA delivery systems for specific therapeutic applications.