DEVELOPMENT AND CHARACTERIZATION OF DOXORUBICIN AND siRNA ENCAPSULATED CHITOSAN NANOPARTICLES
Keywords:Multidrug Resistance (MDR), Chitosan, Doxorubicin, siRNA, Co-Delivery
Objective: Chitosan nanoparticles (ChNP’s) have been widely studied for drug and gene delivery. In this study, we prepared ChNP’s for co-delivery of doxorubicin (DOX) and siRNA for cancer treatment.
Methods: The ionic gelation method was used to develop ChNP’s. The positively charged DOX and negatively charged siRNA encapsulated into ChNP’s. The particle size and zeta potential of the developed ChNP’s were studied by particle size analyzer and morphology was examined by TEM. Encapsulation of DOX in ChNP’s was confirmed by FTIR spectroscopy. The encapsulation efficiency and in vitro release of DOX were studied by UV-Vis spectrophotometry. The siRNA loading into ChNP’s was confirmed by gel retardation assay.
Results: The developed ChNP’s showed particle size ranged from 127±6.5 to 215±8.5 nm with zeta potential ranged from 16.5±0.3 to 25.8±0.3. Transmission Electron Micrograph showed DOX and siRNA encapsulated ChNP’s are polydisperse and spherical in nature. FTIR study confirmed the binding of DOX with ChNP’s with absorption peaks at 1016 cm-1, 1316 cm-1, 1412 cm-1, 1645 cm-1 and 3370 cm-1. The TPP:Ch ratio 0.1:0.5 showed the highest encapsulation efficiency 69±3.24%, with initial burst release and then sustained or slow release of DOX. Agarose gel retardation study confirmed the encapsulation of siRNA in ChNP’s by retarded migration of siRNA-ChNP’s in comparison with naked siRNA.
Conclusion: The developed ChNP’s successfully encapsulated the DOX and siRNA and showed the sustain release of DOX. In conclusion, our study shown that ChNP’s is having a potential of co-loading of DOX-siRNA as an efficient drug delivery system for the treatment of various cancers such as colorectal cancer, breast cancer etc.
Chandran SP, Natarajan SB, Chandrasekaran S, Mohd Shahimi MSB. Nano drug delivery strategy of 5-fluorouracil for the treatment of colorectal cancer. J Cancer Res Practice 2017;4:45-8.
Cisterna BA, Kamaly N, Choi WI, Tavakkoli A, Farokhzad OC, Vilos C. Targeted nanoparticles for colorectal cancer. Nanomedicine (Lond) 2016;11:2443-56.
Kim HJ, Kim A, Miyata K, Kataoka K. Recent progress in the development of siRNA delivery vehicles for cancer therapy. Adv Drug Delivery Rev 2016;104:61-77.
Janes KA, Fresneau MP, Marazuela A, Fabra A, Alonso MaJ. Chitosan nanoparticles as delivery systems for doxorubicin. J Controlled Release 2001;73:255-67.
Nogueira Librelotto DR, Scheeren LE, Vinardell MP, Mitjans M, Rolim CMB. Chitosan-tripolyphosphate nanoparticles functionalized with a pH-responsive amphiphile improved the in vitro antineoplastic effects of doxorubicin. Colloids Surf B 2016;147:326-35.
Souto GD, Farhane Z, Casey A, Efeoglu E, McIntyre J, Byrne HJ. Evaluation of cytotoxicity profile and intracellular localization of doxorubicin-loaded chitosan nanoparticles. Anal Bioanal Chem 2016;408:5443-55.
Hellmers F, Ferguson P, Koropatnick J, Krull R, Margaritis A. Characterization and in vitro cytotoxicity of doxorubicin-loaded γ-polyglutamic acid-chitosan composite nanoparticles. Biochem Eng J 2013;75:72–8.
Butt AM, Amin MC, Katas H, Abdul Murad NA, Jamal R, Kesharwani P. Doxorubicin and siRNA codelivery via chitosan-coated pH-responsive mixed micellar polyplexes for enhanced cancer therapy in multidrug-resistant tumors. Mol Pharm 2016;13:4179-90.
Conde J, de la Fuente JM, Baptista PV. Nanomaterials for reversion of multidrug resistance in cancer: new hope for an old idea? Front Pharmacol 2013;4:134.
Sadreddini S, Safaralizadeh R, Baradaran B, Aghebati Maleki L, Hosseinpour Feizi MA, Shanehbandi D, et al. Chitosan nanoparticles as a dual drug/siRNA delivery system for treatment of colorectal cancer. Immunol Lett 2017;181:79-86.
Bumcrot D, Manoharan M, Koteliansky V, Sah DW. RNAi therapeutics: a potential new class of pharmaceutical drugs. Nature Chem Biol 2006;2:711-9.
Ku SH, Jo SD, Lee YK, Kim K, Kim SH. Chemical and structural modifications of RNAi therapeutics. Adv Drug Delivery Rev 2016;104:16-28.
Lam JK, Chow MY, Zhang Y, Leung SW. siRNA Versus miRNA as therapeutics for gene silencing. Mol Ther Nucleic Acids 2015;4:e252.
Saraswathy M, Gong S. Recent developments in the co-delivery of siRNA and small molecule anticancer drugs for cancer treatment. Materials Today 2014;17:298-306.
Ibrahim H, Farid O, Samir A, Mosaad R. Preparation of chitosan antioxidant nanoparticles as drug delivery system for enhancing of anti-cancer drug. Key Eng Materials 2018;759:92-7.
Calvo P, Remunan Lopez C, Vila Jato JL, Alonso MJ. Novel hydrophilic chitosan-polyethylene oxide nanoparticles as protein carriers. J Appl Polymer Sci 1997;63:125-32.
Sun Q, Wang X, Cui C, Li J, Wang Y. Doxorubicin and anti-VEGF siRNA co-delivery via nano-graphene oxide for enhanced cancer therapy in vitro and in vivo. Int J Nanomed 2018;13:3713-28.
Xu Y, Du Y. Effect of the molecular structure of chitosan on protein delivery properties of chitosan nanoparticles. Int J Pharm 2003;250:215-26.