UJI AKTIVITAS ANTIMALARIA FALCIPARUM NANOPARTIKEL BERBASIS KITOSAN YANG MENGANDUNG ANTISENSE OLIGODEOKSINUKLEOTIDA BERTARGET GEN dhs DAN eba-175

• Main Author: Puji Kusuma D, Dea
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/44574
• Institution: Institut Teknologi Bandung

Malaria infection caused by Plasmodium sp. parasite still has many casualties in the world. In Indonesia, malaria cases still occur in East Indonesia. The main problems of conventional malaria medications are multiple drug resistance and no specific target in parasite’s intracellular. This study tried to develop a new therapy for malaria using asODNs (antisense oligonucleotides) that specifically target to eba-175 and dhs genes of Plasmodium falciparum, that is responsible for the most death. To prevent ODN degradation by nuclease enzymes and to facilitate the internalization of ODN into red blood cells, ODN was packaged into chitosan-based nanoparticles (NPs). Those nanoparticles were chitosan NP, chitosan-poloxamer NP, and chitosan-PLGA NP. Chitosan and chitosan-poloxamer NPs were prepared using ionic gelation method. The sizes of nanoparticles were less than 200 nm with polydispersity index of less than 0.5. Chitosan-PLGA NP was prepared using emulsification solvent diffusion method. The mean size of particles obtained was 561.6 ± 89.3 nm with polydispersity index 0.345 ± 0.044. ODN targeted against dhs loaded chitosan-poloxamer NP showed the highest schizont growth inhibition of approximately 68.1%. Additionally, the inhibition exhibited significantly different (p<0.05) between free ODN and 0.5 µM ODN-loaded chitosan-poloxamer NP. PCR products’ size confirmation was performed to select appropriate primers for PCR-based gene expression assay. Eba-175 f1r1, dhs f4r4, and 18 rRNA f1r1 primers had correct sizes and each primer generated merely one band so that it can be used for PCR-based assays.