FORMULATION AND DENDRITIC CELL MATURATION TEST OF CHITOSAN- HBsAg BASED NANOPARTICLES CONTAINING Moringa oleifera Lam. LEAF EXTRACT AS ADJUVANT

• Main Author: Ika Wijayanti, Rosa
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/37133
• Institution: Institut Teknologi Bandung

The majority of vaccine antigens currently under investigation are composed of highly purified recombinant molecules or subunits of pathogens and as such they lack several features of the pathogens, including the inherent immunostimulatory property, and thus often do not elicit strong immune responses. The purpose of this study was to develop chitosan-based nanoparticle formulations as antigen carriers, HBsAg as an antigen model and extract of Moringa oleifera, Lam. as an adjuvant model to get a better antigen delivery system with the immune response as the parameter. In this study, the ability of Moringa oleifera extract as an adjuvant was tested to improve the immunogenicity of hepatitis B surface antigen (HBsAg). Design of vacine development also plays an important role in achieving increased immune response. This reasearch was to develop HBsAg-loaded nanoparticles and Moringa oleifera aqueous leaf extract as adjuvant by using chitosan polymer. Chitosan nanoparticles was prepared by ionic gelation method using sodium tripolyphosphate as cross-linking agent. A system was composed of chitosan in which HBsAg and M. oleifera extract were incorporated. Concentration of HBsAg used in this combination was 10 µg/mL and concentrations of extract were 10, 50 and 100 µg/mL, respectively. In this study, three types nanoparticles were produced, HBsAg-loaded nanoparticles, M. oleiferaloaded nanoparticles and combination of HBsAg-M. oleifera loaded nanoparticles. The nanoparticles formed were characterized for particle size, polydispersity index, zeta potential, HBsAg entrapment eficiency using silver staining method on SDS-PAGE and extract entrapment efficiency using total flavonoid method. To evaluate the effectiveness of the nanoparticles in stimulating immune response, in vitro dendritic cell maturation study was carried out using flowcytomery with CD86, CD83, CD40 as markers. The results showed that particles size were between 167,58–248,88 nm with a good polydispesity index (<0,5) and exhibited a high positive zeta potential (30,92-37,92). Entrapment efficiency of extract in separated formula was in the range 72,62–86,32 % and entrapment efficiency of HBsAg in formulation was in the range 98,40– 99,63%. The percentage of dendritic cell maturation using all three of labels as markers gave an average above of 50%. In vitro testing showed maturation of dendritic cell in the developed form.