Modulation of Chitosan-TPP Nanoparticle Properties for Plasmid DNA Vaccines Delivery

Area of Science:

• Biotechnology
• Nanomedicine
• Vaccine Development

Background:

• Nucleic acid vaccines require effective delivery systems for viral infections like SARS-CoV-2 and HPV.
• Chitosan (CS) and tripolyphosphate (TPP) are biocompatible polymers suitable for nanoparticle formulation.

Purpose of the Study:

• To develop and characterize chitosan-tripolyphosphate-plasmid DNA (CS-TPP-pDNA) nanoparticles for nucleic acid vaccine delivery.
• To evaluate the stability, cytotoxicity, cellular uptake, and antigen expression of these nanoparticles.

Main Methods:

• Nanoparticle characterization using dynamic light scattering (DLS) for size and charge, and scanning electron microscopy (SEM) for morphology.
• In vitro evaluation of nanoparticle stability, cytotoxicity, and cellular uptake.
• Assessment of HPV E7 antigen expression using polymerase chain reaction (PCR) and real-time PCR.

Main Results:

• CS-TPP-pDNA nanoparticles exhibited spherical/oval shapes, sizes <180 nm, and positive zeta potentials (>20 mV).
• Nanoparticles demonstrated good stability in storage and physiological conditions, with improved cell viability upon buffer elimination.
• Increased E7 antigen transcription was observed with higher pDNA concentrations (60 μg/mL).

Conclusions:

• CS-TPP-pDNA polyplexes represent a promising and stable vehicle for nucleic acid vaccine delivery.
• This nanoparticle system shows potential for preventing and treating viral infections, including those caused by emergent pathogens.