Stable Polymer-Lipid Hybrid Nanoparticles Based on mcl-Polyhydroxyalkanoate and Cationic Liposomes for mRNA Delivery

Area of Science:

• Biotechnology
• Materials Science
• Nanotechnology

Background:

• Polymer-lipid hybrid nanoparticles (PLNs) offer improved stability for cationic lipid carriers.
• PLNs combine advantages of polymer nanoparticles and liposomes, enhancing physical stability and biocompatibility.
• Natural polymers like polyhydroxyalkanoate (PHA) can serve as matrices for PLN preparation.

Purpose of the Study:

• To develop stable cationic hybrid PLNs using a specific cationic lipid (2X3), DOPE, and mcl-PHA.
• To evaluate the role of mcl-PHA in enhancing the stability of PLNs in ionic solutions.
• To assess the mRNA-eGFP delivery efficiency of the developed PLNs to BHK-21 cells.

Main Methods:

• Synthesis of cationic hybrid PLNs using 2X3, DOPE, and mcl-PHA from *Pseudomonas helmantisensis* P1.
• Encapsulation and delivery of mRNA-eGFP to BHK-21 cells.
• Stability assessment in phosphate-buffered saline (PBS) using dynamic light scattering, electrophoretic mobility, and transmission electron microscopy.

Main Results:

• The developed PLNs successfully encapsulated and delivered mRNA-eGFP to BHK-21 cells.
• Cationic lipid carriers (CLs) showed decreased stability and aggregation in increasing PBS concentrations.
• PLNs demonstrated stability for 120 hours at +4 °C in isotonic PBS, without aggregation.

Conclusions:

• The hydrophobic polymer mcl-PHA significantly enhances the stability of cationic PLNs in ionic environments.
• PLNs offer a superior alternative to CLs for nucleic acid delivery in the presence of physiological salt concentrations.
• The developed PLNs show considerable potential for future in vivo nucleic acid delivery studies.