APPLICATION OF BOX-BEHNKEN DESIGN IN OPTIMIZATION OF SURFACE MODIFIED CHITOSAN-ACEMANNAN LIPID NANOPARTICLES AS RIFAMPICIN DELIVERY SYSTEM

• Main Author: Soraya Choirunissa, Eldi
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/44495
• Institution: Institut Teknologi Bandung

Intracelullar infections become important public health concern due to persistent and difficult to treat. Various pathogenic bacteria such as Staphylococcus aureus (SA) and Mycobacterium tuberculosis (Mtb) invade targeted cells, which are able to escape from phagolysosomal degradation and persist within cytoplasm for prolonged periods. Unfortunately, many antibiotics fail to accumulate efficiently and exhibit poor retention inside the cells. The goal of this study is to develop an active intracellular targeting vehicle of rifampicin in the form of surface modified lipid nanoparticle formulation (ACE-NP) by chitosan-acemannan conjugate (COS-ACE) using Box-Behnken design. The nanoparticles were prepared using emulsification-ultrasonication method by mixing oil phase of lipid and rifampicin solution and aqueous solution containing COS-ACE and polisorbate 80. Subsequently, the particles ACE-NP was cross-linked with sodium tripolyphosphate for gelation ionotropic solidification. The formulation was optimized using Box-Behnken design with the dependent variables of lipid, COS-ACE, and polisorbate 80 concentrations and the responses of particle size, entrapment efficiency (EE) and drug loading (DL). The optimum formula lied at the center point of the model, which consisted of 0.75% lipid; 0.75% COS-ACE; 5% polisorbate 80. The formula showed particle size of 416.14 ± 60.16 nm, EE of 88.51 ± 0.21% and DL of 35.46 ±0.09 % with % errors of 0.72; 0.03 and 0.16, respectively. This ACE-NP had zeta potential value of +1.66 mV and a similar release profile of rifampicin ACE-NP at pH 5.2 and 7.4. The minimum inhibition and bactericidal concentration of ACE-NP was equivalent with those of RIF solution concentration at 25 µg/mL. Importantly, based on qualitative assay, the ACE-NP was shown no-obvious nucleus cell translocation within Vero cells and 3t3 Balb/C cells. This study indicates that the optimum formula obtained in Box-Behnken design is potential candidate for delivering intracellular targeting of RIF as antibiotics.