Development, characterization, and in vitro evaluation of phosphatidylcholine–sodium cholate-based nanoparticles for siRNA delivery to MCF-7 human breast cancer cells
- 206 Downloads
Phosphatidylcholine–sodium cholate (SC)-based nanoparticles were designed, characterized, and evaluated as plausible oligonucleotides delivery systems. For this purpose, formulation of the systems was optimized to obtain low cytotoxic vehicles with high siRNA-loading capacity and acceptable transfection ability. Mixtures of soybean phosphatidylcholine (SPC) and SC were prepared at different molar ratios with 2 % w/v total concentration; distilled water and two different buffers were used as dispersion medium. Nanoparticles below 150 nm were observed showing spherical shape which turned smaller in diameter as the SC molar proportion increased, accounting for small unilamellar vesicles when low proportions of SC were present in the formulation, but clear mixed micellar solutions at higher SC percentages. Macroscopic characteristics along with physico-chemical parameters values supported the presence of these types of structures. SYBR green displacement assays demonstrated an important oligonucleotide binding that increased as bile salt relative content got higher. Within the same molar ratio, nanoparticles showed the following binding efficiency order: pH 7.4 > pH 5.0 > distilled water. siRNA-loading capacity assays confirmed the higher siRNA binding by the mixed micelles containing higher SC proportion; moreover, the complexes formed were smaller as the SC:SPC ratio increased. Considering cytotoxicity and siRNA-loading capacity, 1:2 and 1:4 SPC:SC formulations were selected for further biological assays. Nanoparticles prepared in any of the three media were able to induce dsRNA uptake and efficiently transfect RNA for gene silencing, for the compositions prepared in buffer pH 5.0 being the most versatile.
KeywordssiRNA delivery system Antisense therapy Phosphatidylcholine–sodium cholate-based nanoparticles Gene knockdown Nanomedicine
Support for these studies was provided by the National Agency of Scientific and Technological Promotion (ANPCyT), Ministry of Science, Technology, and Productive Innovation, Argentina, the University of Buenos Aires, and the National Science Research Council (CONICET). The authors have no relevant financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript. No writing assistance was utilized in the production of this manuscript.
- Cabral DJ, Hamilton JA, Small DM (1986) The ionization behavior of bile acids in different aqueous environments. J Lipid Res 27:334–344Google Scholar
- Dorasamy S, Moganavelli S, Ariatti M (2009) Rapid and sensitive fluorometric analysis of novel galactosylated cationic liposome interaction with siRNA. Afr J Pharm Pharmacol 3:632–635Google Scholar
- Dürr M, Hager J, Lohr JP (1994) Investigation on mixed micelle and liposome preparations for parenteral use based on soya phosphatidylcholine. Eur J Pharm Biopharm 40:147–156Google Scholar
- Gándola Y, Pérez SE, Irene PE, Sotelo AI, Miquet JG, Corradi GR, Carlucci AM, González L (2014) Mitogenic effects of phosphatidylcholine nanoparticles on MCF-7 breast cancer cells. Biomed Res Int. Vol 2014, Article ID 687037, p 13. doi: 10.1155/2014/687037
- Hendradi E, Obata Y, Takayama K, Nagai T (2003) Effect of bile salts-lecithin mixed micelles on the skin permeation of diclofenac in rats. STP Pharm Sci 13(4):247–251Google Scholar
- Lemke TL (2008) Pharmaceutical biotechnology: from nucleic acids to personalized medicine. In: (ed) Foye’s principles of medicinal chemistry, 6th edn. Lippincott Williams & Wilkins, Philadelphia, pp 115–175Google Scholar
- Manjunath N, Dykxhoorn DM (2010) Advances in synthetic siRNA delivery. Discov Med 9(48):418–430Google Scholar
- Monteagudo E, Gándola Y, González L, Bregni C, Carlucci A (2012) Development, characterization and in vitro evaluation of tamoxifen microemulsions. J Drug Deliv. Vol 2012, Article ID 236713, p 11. doi: 10.1155/2012/236713
- Modi P (2000) Mixed micellar delivery system and method of preparation. US Patent 6017545 A, 25 January 2000Google Scholar
- Pérez SE, Gándola Y, Carlucci AM, González L, Turyn D, Bregni C (2012) Formulation strategies, characterization, and in vitro evaluation of lecithin-based nanoparticles for siRNA delivery. J Drug Deliv. Vol 2012, Article ID 986265, p 9, doi: 10.1155/2012/986265