Developing a Ready-to-Use Lipid Nanoparticle Technology for Nucleic Acid Delivery Based on Deep Eutectic Solvents.

Area of Science:

• Biotechnology
• Materials Science
• Drug Delivery

Background:

• Microfluidic technology is a common method for producing lipid nanoparticles (LNPs) for nucleic acid delivery.
• The widespread use of microfluidics is limited by specialized, expensive equipment and consumables.
• Current methods often rely on organic solvents, posing cost and safety concerns.

Purpose of the Study:

• To develop a novel, cost-effective, and user-friendly technology for producing lipid nanoparticles (LNPs).
• To create ready-to-use lipid nanoparticles (RULNPs) using deep eutectic solvents (DESs) as an alternative to microfluidics.
• To evaluate the physicochemical properties, delivery efficacy, and cellular uptake of DES-based RULNPs.

Main Methods:

• A deep eutectic solvent (DES) composed of fructose and glycerol ([Fru][Gly]) was utilized.
• Lipids and nucleic acids were dissolved in the DES, facilitating RULNP formation through simple physical mixing and hydration.
• Physicochemical properties and in vitro delivery efficacy of RULNPs were compared to traditional LNPs.

Main Results:

• The [Fru][Gly] DES effectively dissolved lipids and nucleic acids, enabling the formation of RULNPs.
• RULNPs demonstrated comparable physicochemical properties and nucleic acid (plasmid DNA and RNA) delivery efficacy to LNPs.
• Mechanistic studies indicated superior cellular uptake of RULNPs compared to LNPs, despite comparable endosomal escape.

Conclusions:

• Deep eutectic solvent-based RULNP technology offers a rapid, straightforward, and cost-effective alternative for LNP production.
• This innovative approach circumvents the limitations of microfluidic methods, including high costs and reliance on organic solvents.
• DES-based RULNPs hold significant potential for revolutionizing nucleic acid delivery applications.