Folate receptor mediated intracellular gene delivery using the charge changing solid lipid nanoparticles

Area of Science:

• Biomaterials Science
• Nanotechnology
• Gene Therapy

Background:

• Non-viral gene delivery systems offer improved safety over viral vectors but suffer from low transfection efficiency.
• Folic acid receptor-mediated endocytosis presents a strategy to enhance cellular uptake and transfection rates.
• Developing effective non-viral carriers is crucial for advancing gene therapy applications.

Purpose of the Study:

• To synthesize and characterize novel charge-changing Solid Lipid Nanoparticles (Folate-chitosan-CHETA-Sln) for enhanced gene delivery.
• To evaluate the targeting capability and transfection efficiency of these nanoparticles in folate receptor-positive and -negative cells.
• To assess the biocompatibility and stability of the developed gene delivery system.

Main Methods:

• Folate-chitosan and cholesterol derivative CHETA were synthesized and used to create Folate-chitosan-CHETA-Sln nanoparticles via a reverse micelle-double emulsion method.
• Particle size, zeta potential, and morphology were analyzed using dynamic light scattering and scanning electron microscopy (SEM).
• Transfection efficiency was evaluated in SKOV3 (folate receptor-overexpressing) and A549 (folate receptor-deficient) cell lines, with cytotoxicity assessed in HEK 293 cells.

Main Results:

• Folate-chitosan-CHETA-Sln nanoparticles exhibited a size of 254.5 ± 20 nm and an initial zeta potential of -40.5 ± 0.8 mV, with a spherical morphology observed via SEM.
• Treatment with dithiothreitol (DTT) reversed the zeta potential from negative to positive (20.5 ± 1.9 mV), indicating charge-changing properties.
• Enhanced reporter gene expression was observed in SKOV3 cells compared to A549 cells, demonstrating targeted delivery. The system showed no significant cytotoxicity in HEK 293 cells and maintained transfectivity in the presence of serum.

Conclusions:

• Folate-chitosan-CHETA-Sln nanoparticles possess favorable physical characteristics and demonstrate high transfection efficiency, particularly in targeting folate receptor-expressing cells.
• The charge-changing capability and stability in serum suggest potential for effective non-viral gene delivery.
• These nanoparticles represent a promising novel system for gene delivery applications, balancing biocompatibility with enhanced transfection rates.