Zeta potential changing phosphorylated nanocomplexes for pDNA delivery.

Area of Science:

• Biomaterials Science
• Gene Therapy
• Polymer Chemistry

Background:

• Gene delivery systems are crucial for therapeutic applications.
• Polyethyleneimine (PEI) is a widely used non-viral gene vector.
• Modifying PEI can enhance its gene delivery efficiency and safety.

Purpose of the Study:

• To evaluate a zeta potential changing system utilizing modified polyethyleneimine (PEI) as a gene delivery vector.
• To investigate the attachment of 6-phosphogluconic acid (6-PGA) to linear and branched PEI.
• To assess the impact of zeta potential modification on gene delivery efficacy.

Main Methods:

• Synthesized PEI-6-phosphogluconic acid (PEI-6-PGA) conjugates via carbodiimide chemistry.
• Formed nanocomplexes with plasmid DNA (pDNA) and modulated zeta potential using carboxymethylcellulose (CMC).
• Evaluated phosphate ester cleavage in Caco-2 and HEK-293 cells and measured green fluorescent protein (GFP) expression for transfection efficiency.

Main Results:

• PEI-6-PGA nanocomplexes exhibited tunable zeta potentials, reduced by CMC and increased by alkaline phosphatase.
• Enzymatic degradation of the phosphate ester was higher in Caco-2 cells compared to HEK-293 cells.
• Transfection efficiency, indicated by GFP expression, was higher in HEK-293 cells and decreased with phosphate inhibitors.

Conclusions:

• Zeta potential changing systems represent a promising strategy for advanced gene delivery.
• In situ generation of positive charges near the cell surface enhances gene delivery.
• The PEI-6-PGA system demonstrates potential for targeted and efficient gene therapy applications.