DEVELOPMENT OF A KIDNEY TARGETTED NANOCARRIER FORMULATION AGAINST CISPLATIN INDUCED NEPHROTOXICITY

Abstract

Chitosan is a natural polymer which is less toxic in the body than other synthetic polymers. Chitosan sodium tripolyphosphate (TPP) nanoparticles produced by ionic gelation method are known to be good drug delivery agents for human body tissues, such as brain and kidney. In this study, to eliminate nephrotoxicity in kidney caused by cisplatin, an anti-cancer drug, chitosan-TPP nanoparticles loaded with gene silencing siRNAs were used. In vitro studies have been tested in human kidney cell line Hek293 cells and the toxic effects of nanoparticles were found to be low compared to 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide and 2,3- Bis-(2-Methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide, disodium salt cell cytotoxicity results. The penetration of nanoparticles into the cell was confirmed by fluorescence microscopy and flow cytometry. In vivo studies have shown that nanoparticle injected with siRNA loaded into rats of type 6-8 week in the Balb-c type. Control and siRNA-loaded chitosan nanoparticles cisplatin-treated mice group were used for the animal experiment. After cisplatin injections, creatinine and BUN assays were performed to detect the level changing by nanoparticles after cisplatin-induced siRNA delivery. GAPDH is a control gene and PKCδ, P53, OCT1, OCT2 and γGT genes have important roles in kidney proximal tubule cells. In this study, mRNA levels of these genes were also examined by quantitative PCR. While siRNAs were decreased in the first days of injection, this effect was lost in the following days. Thus, the silencing potential of each siRNA is variable. However, this variability shows a significant change in the study.