Preparation of CaP/pDNA nanoparticles by reverse micro-emulsion method: Optimization of formulation variables using...

Area of Science:

• Biomaterials Science
• Nanotechnology
• Gene Delivery

Background:

• Developing effective non-viral gene delivery vectors is crucial for gene therapy.
• Calcium phosphate (CaP) nanoparticles offer potential due to their biocompatibility and biodegradability.
• Plasmid DNA (pDNA) is a common cargo for gene delivery applications.

Purpose of the Study:

• To optimize the preparation of calcium phosphate/plasmid DNA (CaP/pDNA) nanoparticles.
• To characterize the physical and functional properties of the optimized nanoparticles.
• To evaluate the gene transfection efficiency of the developed CaP/pDNA nanoparticles.

Main Methods:

• Preparation of CaP/pDNA nanoparticles using a Triton X-100/Butanol/Cyclohexane/Water reverse microemulsion system.
• Optimization of preparation parameters using the Box-Behnken design method based on particle size.
• Characterization of particle size, polydispersity index, and pDNA encapsulation efficiency.

Main Results:

• Optimized CaP/pDNA nanoparticles achieved a size of 60.23 ± 4.72 nm with a polydispersity index of 0.252.
• pDNA encapsulation efficiency exceeded 90% in the optimized nanoparticles.
• The optimized nanoparticles exhibited pH sensitivity, biocompatibility, and higher transfection efficiency compared to unoptimized formulations.

Conclusions:

• The reverse microemulsion method, optimized via Box-Behnken design, effectively produces small, highly encapsulated CaP/pDNA nanoparticles.
• The optimized CaP/pDNA nanoparticles possess desirable properties including pH sensitivity and biocompatibility for gene delivery.
• These nanoparticles represent a promising non-viral vector for enhanced gene transfection applications.