Preparation of N-Trimethyl Chitosan-Protein Nanoparticles Intended for Vaccine Delivery

Abstract

In this paper, various N-trimethyl chitosan (TMC) of different molecular-weights (approximately 100 KD, approximately 200 KD, and approximately 400 KD, respectively) with the approximately degree of quartenization (DQ) of 40% were successfully synthesized. In vitro cytotoxicity of TMC solution showed the dependence of TMC concentration from 20 microg/ml to 500 microg/ml on the relative cell activity. Molecular weight of TMC did not greatly affect the cytotoxicity of TMC against HEK293 and L929 cells. TMC nanoparticles and alginate modified TMC nanoparticles were prepared by the ionic gelation method. Subsequently, we investigated the properties of TMC nanoparticles and alginate modified TMC nanoparticles intending for oral delivery of antigens. Molecular weight of TMC did not affect the loading capacity (LC) and in vitro release behavior of TMC nanoparticles. However, BSA concentration and alginate modification have strongly effect on properties of TMC nanoparticles (particle size; surface charge; loading efficiency and loading capacity). In vitro release behavior indicated that alginate modification could efficiently decrease initial burst release and extend release time in phosphate buffer (PBS, pH 7.4) and acidic solution (0.1 M HCl, pH = 1) at 37 degrees C. SDS-polyacrylamide gel electrophoresis (SDS-PAGE) assay showed that alginate modification could effectively improve the stability of TMC nanoparticles and protect BSA from degradation or hydrolysis in acidic condition for at least 2 h.