Synthesis and characterization of Fe₃O₄-PEG-LAC-chitosan-PEI nanoparticle as a survivin siRNA delivery system

Abstract

<jats:p> The limited effectiveness of the conventional methods for cancer treatment makes the researchers to find novel safe and effective therapeutic strategies. One of these strategies is to use small interfering RNAs (siRNAs). A major challenge here is the siRNA delivery into the cells. The purpose of this study was to design and prepare a biocompatible, biodegradable, and safe nanosized particle for siRNA delivery into human breast cancer MCF-7 and leukemia K562 cells. Chemically synthesized magnetic nanoparticles containing polyethyleneglycol-lactate polymer (PEG-LAC), chitosan, and polyethyleneimine (PEI) were successfully prepared and used as a gene delivery vehicle. The nanoparticles were characterized by Fourier transform infrared spectroscopy and zeta potential. The Fe<jats:sub>3</jats:sub>O<jats:sub>4</jats:sub>-PEG-LAC-chitosan-PEI nanoparticle showed efficient and stable survivin siRNA loading in gel retardation assay. The cytotoxicity of the prepared nanoparticle was studied using 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay and was compared with that of mitoxantrone (MTX) in combination with the prepared siRNA delivery system to evaluate the possible synergic effect of MTX and survivin siRNA. The nanoparticles with and without noncomplementary siRNA showed low toxicity against both cell lines; however, a twofold decrease was observed in cell survival percent after MTX addition to MCF-7 cells treated with either nanoparticle itself or complexed with noncomplementary siRNA. While survivin siRNA nanoplex caused threefold decrease in the cell survival percent, its combination with MTX did not result in a significant increase in the cytotoxic effect. Therefore, Fe<jats:sub>3</jats:sub>O<jats:sub>4</jats:sub>-PEG-LAC-chitosan-PEI nanoparticle should be considered as a potential carrier for enhanced survivin siRNA delivery into MCF-7 and K562 cells. </jats:p>

Chitosan as possible inhibitory agents and delivery systems in leukemia

“…In recent years, the use of RNA interference in gene therapy methods has been widely welcomed by researchers. However, one of the major problems with this treatment is the transfer of small interfering RNAs (siRNAs) to target tumor cells [158][159][160][161][162][163][164]. Various pathways are involved in the inhibition or progression of cancer in the body, the most important of which are cell death and survival.…”

“…This gene is highly expressed in cancer cells. Reports indicate that if survivin expression is suppressed, cancer cells become sensitive to anticancer compounds and drugs [158,165,166]. From these findings, it can be concluded that inhibiting Survivin expression is a potentially valuable target for cancer treatment and suppression of its progression.…”

“…In the synthesis of these carrier nanoparticles, PEG reduces PEI toxicity and ensures the stability of colloidal particles. On the other hand, PEG, in addition to its high biocompatibility, prevents the deposition of nanoparticles [158,173]. Arami et al [158] designed the "Fe3O4-PEG-LAC-chitosan-PEI" nanoparticle carrier, which due to the properties of the compounds used in it, has a sufficient positive charge to react with siRNAs [158].…”

“…On the other hand, PEG, in addition to its high biocompatibility, prevents the deposition of nanoparticles [158,173]. Arami et al [158] designed the "Fe3O4-PEG-LAC-chitosan-PEI" nanoparticle carrier, which due to the properties of the compounds used in it, has a sufficient positive charge to react with siRNAs [158]. In their study, they used the nanoparticle to transfer survivin siRNA to human breast cancer cells (MCF-7) and human chronic myelogenous leukemia cells (K562) in vitro.…”

Nanoparticles as Theranostic Vehicles in Experimental and Clinical Applications—Focus on Prostate and Breast Cancer

“…The protein survivin, encoded by the BIRC5 oncogene in the human genome, and which inhibits the caspase activation and therefore downregulates the apoptotic pathway, has received much attention lately. Several attempts, also including NPs, have been made to distribute anti-survivin siRNA in order to silence the BIRC5 gene [445,446,447]. For NP-based GTH applications for PCa, there are also quite few publications the last decade [141,448,449,450,451,452].…”