PENGARUH VARIASI PH PADA FORMULASI TERHADAP KARAKTERISTIK NANOPARTIKEL KITOSAN SEBAGAI AGEN PENGENKAPSULASI PLASMID PIMSY

• Main Author: Hanifah Ramadhani, Nur
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/80828
• Institution: Institut Teknologi Bandung

Infectious Myonecrosis (IMN), a disease caused by the pathogen Infectious Myonecrosis Virus (IMNV), causes an increase in shrimp mortality rates ranging from 30% to 100%. Invertebrates such as shrimp do not have adaptive immunity, so the development of a vaccine to prevent IMNV is not possible. An alternative is the RNA interference (RNAi) method. In the delivery process, oral exposure is a suitable approach in shrimp farming because it is easy and more efficient in terms of time and cost. However, in this delivery system, plasmids containing dsRNA sequences cannot be exposed directly because they are less stable and easily degraded by various enzymatic activities. One of the delivery mechanisms that can be used is plasmid encapsulation using nanoparticle technology based on natural polymers such as chitosan. One of the factors that affect the results of the physical and chemical characteristics of chitosan nanoparticles is pH, which will also affect the interaction of nanoparticles with cells. Therefore, this study aims to compare the characteristics of chitosan nanoparticles formulated with pH variations of 3.5, 4.5, and 5.5 and determine the best pH variation to be used in encapsulating pIMSY plasmids containing RNAi sequences. Nanoparticles were formed by ionic gelation method followed by tests including characterization: indirect encapsulation efficiency, plasmid stability, size (Z-average), polydispersity index as well as TEM image visualization, zeta potential, and in vitro release. The results showed that the pH parameter in the encapsulation process has an influence on the strength of cross-linking between chitosan and TPP which in turn causes differences in the characteristics: encapsulation efficiency; Z-average size; zeta potential; and in vitro release of plasmids from chitosan nanoparticles. Based on this study, the most optimum formulation pH is at pH 4.5 variation which has a profile in the form of z-average size 113.87 nm; polydispersity index 0.49; zeta potential 0.2 mV; encapsulation efficiency 71.91%; and on the 18th day of the cumulative in vitro release was 9.1%.