Chitosan-DNA nanoparticles: synthesis and optimization for long-term storage and effective delivery.

Area of Science:

• Biotechnology
• Nanotechnology
• Gene Delivery

Background:

• Chitosan nanoparticles (CsNPs) are cost-effective for DNA delivery.
• Current CsNP synthesis methods lack optimized long-term storage solutions.
• This study addresses CsNP storage and functional stability.

Purpose of the Study:

• To develop and optimize a protocol for CsNP synthesis, lyophilization, and storage.
• To maintain CsNP transfection efficiency after long-term storage.
• To enable broader applications of CsNP technology.

Main Methods:

• CsNP synthesis and characterization using DLS, TEM, and zeta potential measurements.
• HEK293 cell transfection with CsNPs and gene expression analysis via flow cytometry and confocal microscopy.
• Optimization of lyophilization and sonication parameters for CsNPs.

Main Results:

• Optimal N:P ratios (2:1 and 3:1) yielded smaller CsNPs with high transfection efficiency.
• Lyophilized CsNPs lost 50% efficiency, but sonication restored it to fresh CsNP levels.
• Sonicated CsNPs maintained spherical morphology and showed minimal cytotoxicity (>90% viability).

Conclusions:

• An optimized protocol for CsNP synthesis, lyophilization, and sonication was established.
• This method allows for long-term storage of CsNPs while preserving transfection efficiency.
• The findings facilitate wider use of CsNPs in gene delivery applications.