Synthesis and characterization of chitosan nanoparticles: Insights from in-vitro analysis

Chitosan nanoparticles (CNPs) are widely acknowledged for their versatility in various domains, particularly in areas relevant to pharmaceuticals and vaccines. This study aimed to produce CNPs as a foundational element for nanoparticle-based delivery systems intended for potential use in vaccine development. We outline a straightforward yet effective synthesis method for generating high-yield, uniform CNPs ranging in size from 100 to 250 nm. The synthesis involved ionic gelation using medium molecular weight chitosan (with a deacetylation level exceeding 75%) and sodium tripolyphosphate as a crosslinker. This was followed by thorough homogenization to ensure consistency, and purification via 0.45 µm polyethersulfone syringe filters. The CNPs underwent characterization for zeta size, zeta charge, and polydispersity index (PDI), while scanning transmission electron microscopy was employed for morphological analysis. Internalization of these chitosan nanoparticles into MDBK cells was achieved through the use of FITC-conjugated counterparts (FCNP). Furthermore, we investigated the impact of chitosan concentrations on CNP size, charge, and polydispersity. These synthesized CNPs exhibit a strong affinity for proteins, DNA, and other biomolecules, positioning them as potential candidates for the development of nanoparticle-based delivery systems tailored to vaccine development.