Skip to main content

Advertisement

SpringerLink
Log in

Development, characterization, and in vitro evaluation of phosphatidylcholine–sodium cholate-based nanoparticles for siRNA delivery to MCF-7 human breast cancer cells

  • Research Paper
  • Published:
Journal of Nanoparticle Research Aims and scope Submit manuscript

Abstract

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

References

  • Barenholz Y (2012) Doxil®—The first FDA-approved nano-drug: lessons learned. J Control Release 160:117–134

    Article  Google Scholar 

  • Bosselmann S, Williams RO (2012) Has nanotechnology led to improved therapeutic outcomes? Drug Dev Ind Pharm 38:158–170

    Article  Google Scholar 

  • Cabral DJ, Hamilton JA, Small DM (1986) The ionization behavior of bile acids in different aqueous environments. J Lipid Res 27:334–344

    Google Scholar 

  • Chen H, Guo Z, Yu F, Ki J et al (2008) Influence of La3+ ions on the egg-yolk phosphatidylcholine and sodium taurocholate self-assemblies in aqueous suspension. J Colloid Interface Sci 328:158–165

    Article  Google Scholar 

  • Cukierman E, Khan DR (2010) The benefits and challenges associated with the use of drug delivery systems in cancer therapy. Biochem Pharmacol 80:762–770

    Article  Google Scholar 

  • Dangi JS, Vyas SP, Dixit VK (1998) Effect of various lipid-bile salt mixed micelles on the intestinal absorption of Amphotericin-B in rat. Drug Dev Ind Pharm 24(7):631–635

    Article  Google Scholar 

  • de Fougerolles A, Vornlocher HP, Maraganore J, Lieberman J (2007) Interfering with disease: a progress report on siRNA-based therapeutics. Nat Rev Drug Discov 6(6):443–453

    Article  Google Scholar 

  • Dokka S, Toledo D, Shi X, Castranova V, Rojanasakul Y (2000) Oxygen radical-mediated pulmonary toxicity induced by some cationic liposomes. Pharm Res 17:521–525

    Article  Google 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 3:632–635

    Google 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 40:147–156

    Google Scholar 

  • Fisher TL, Terhorst T, Cao X, Wagner RW (1993) Intracellular disposition and metabolism of fluorescently-labeled unmodified and modified oligonucleotides microinjected into mammalian cells. Nucleic Acids Res 21:3857–3865

    Article  Google 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

  • Geall AJ, Blagbrough IS (2000) Rapid and sensitive ethidium bromide fluorescence quenching assay of polyamine conjugate-DNA interactions for the analysis of lipoplex formation in gene therapy. J Pharm Biomed Anal 22:849–859

    Article  Google Scholar 

  • Gomes-da-Silva LC, Fonseca NA, Moura V, Pedroso de Lima MC, Simões S, Moreira JN (2012) Lipid-based nanoparticles for siRNA delivery in cancer therapy: paradigms and challenges. Acc Chem Res 45(7):1163–1171. doi:10.1021/ar300048p

    Article  Google Scholar 

  • Gooding M, Browne LP, Quinteiro FM, Selwood DL (2012) siRNA delivery: from lipids to cell-penetrating peptides and their mimics. Chem Biol Drug Des 80:787–809

    Article  Google Scholar 

  • Grobmyera SR, Zhoua G, Gutweina LG, Iwakumab N, Sharmac P, Hochwald SN (2012) Nanoparticle delivery for metastatic breast cancer. Nanomedicine 8:S21–S30

    Article  Google Scholar 

  • Günther M, Lipka J, Malek A, Gutsch D, Kreyling W et al (2011) Polyethylenimines for RNAi-mediated gene targeting in vivo and siRNA delivery to the lung. Eur J Pharm Biopharm 77:438–449

    Article  Google Scholar 

  • Guo X, Huang L (2012) Recent advances in nonviral vectors for gene delivery. Acc Chem Res 45(7):971–979. doi:10.1021/ar200151m

    Article  Google Scholar 

  • Hammad MA, Müller BW (1998) Increasing drug solubility by means of bile salt–phosphatidylcholine-based mixed micelles. Eur J Pharm Biopharm 46(3):361–367

    Article  Google Scholar 

  • 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):247–251

    Google Scholar 

  • Judge AD, Robbins M, Tavakoli I, Levi J, Hu L et al (2009) Confirming the RNAi-mediated mechanism of action of siRNA-based cancer therapeutics in mice. J Clin Invest 119(3):661–673

    Article  Google Scholar 

  • Kapoor M, Burgess DJ, Patil SD (2012) Physicochemical characterization techniques for lipid based delivery systems for siRNA. Int J Pharm 427:35–57

    Article  Google Scholar 

  • Katas H, Oya Alpar H (2006) Development and characterization of chitosan nanoparticles for siRNA delivery. J Control Release 115(2):216–225

    Article  Google Scholar 

  • Khurana B, Goyal AK, Budhiraja A, Arora D, Vyas SP (2010) siRNA delivery using nanocarriers—an efficient tool for gene silencing. Curr Gene Ther 10(2):139–155

    Article  Google Scholar 

  • Lammers T, Kiessling F, Hennink WE, Storm G (2012) Drug targeting to tumors: principles, pitfalls and (pre-) clinical progress. J Control Release 161:175–187

    Article  Google 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 115–175

  • Lv H, Zhang S, Wang B, Cui S, Yan J (2006) Toxicity of cationic lipids and cationic polymers in gene delivery. J Control Release 114(1):100–109

    Article  Google Scholar 

  • Manjunath N, Dykxhoorn DM (2010) Advances in synthetic siRNA delivery. Discov Med 9(48):418–430

    Google 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

  • Müller K (1981) Structural dimorphism of bile salt/lecithin mixed micelles. A possible regulatory mechanism for cholesterol solubility in bile? X-ray structure analysis. Biochemistry 20:404–414

    Article  Google Scholar 

  • Naik S, Chougule M, Padhi BK, Misra A (2005) Development of novel lyophilized mixed micelle amphotericin B formulation for treatment of systemic fungal infection. Curr Drug Deliv 2(2):177–184

    Article  Google Scholar 

  • Nishikawa M, Huang L (2001) Nonviral vectors in the new millennium: delivery barriers in gene transfer. Hum Gene Ther 12(8):861–870

    Article  Google Scholar 

  • Omedes Pujol M, Coleman DJL, Allen CD, Heidenreich O, Fulton DA (2013) Determination of key structre-activity relationships in siRNA delivery with a mixed micelle system. J Control Release 172(3):939–945

    Article  Google Scholar 

  • Ozpolat B, Sood AK, Lopez-Berestein G (2010) Nanomedicine based approaches for the delivery of siRNA in cancer. J Intern Med 267(1):44–53

    Article  Google Scholar 

  • Modi P (2000) Mixed micellar delivery system and method of preparation. US Patent 6017545 A, 25 January 2000

  • 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

  • Schroeder A, Levins CG, Cortez C, Langer R, Anderson DG (2010) Lipid-based nanotherapeutics for siRNA delivery. J Intern Med 267(1):9–21

    Article  Google Scholar 

  • Shim MS, Kwon YJ (2010) Efficient and targeted delivery of siRNA in vivo. FEBS J 277(23):4814–4827

    Article  Google Scholar 

  • Smidt JH, Grit M, Crommelin DJA (1994) Dissolution kinetics of griseofulvin in mixed micellar solutions. J Pharm Sci 83:1209–1212

    Article  Google Scholar 

  • Spagnou S, Miller AD, Keller M (2004) Lipidic carriers of siRNA: differences in the formulation, cellular uptake, and delivery with plasmid DNA. Biochemistry 43(42):13348–13356

    Article  Google Scholar 

  • Sun Y, Miguéliz I, Navarro G, Tros de Ilarduya C (2009) Structural and morphological studies of cationic liposomes-DNA complexes. Lett Drug Des Discov 6:33–37

    Article  Google Scholar 

  • Sznitowska M, Klunder M, Placzek M (2008) Paclitaxel solubility in aqueous dispersions and mixed micellar solutions of lecithin. Chem Pharm Bull 56(1):70–74

    Article  Google Scholar 

  • Tan Y, Qi J, Lu Y, Hu F, Yin Z, Wu W (2013) Lecithin in mixed micelles attenuates the cytotoxicity of bile salts in Caco-2 cells. Toxicol In Vitro 27(2):714–720

    Article  Google Scholar 

  • Venneman NG, Huisman SJ, Moschetta A, van Berge-Henegouwen GP, van Erpecum KJ (2002) Effects of hydrophobic and hydrophilic bile salt mixtures on cholesterol crystallization in model biles. Biochim Biophys Acta 1583(2):221–228

    Article  Google Scholar 

  • Walter A, Vinson P, Kaplun A, Talmon Y (1991) Intermediate structures in the cholate-phosphatidylcholine vesicle-micelle transition. Biophys J 60:1315–1325

    Article  Google Scholar 

  • Walter A, Kuehl G, Barnes K, VanderWaerdt G (2000) The vesicle-to-micelle transition of phosphatidylcholine vesicles induced by nonionic detergents: effects of sodium chloride, sucrose and urea. Biochim Biophys Acta 1508:20–33

    Article  Google Scholar 

  • Wu SY, Singhania A, Burgess M et al (2011) Systemic delivery of E6/7 siRNA using novel lipidic particles and its application with cisplatin in cervical cancer mouse models. Gene Ther 18(1):14–22

    Article  Google Scholar 

  • Zamora MR, Budev M, Rolfe M, Gottlieb J, Humar A et al (2011) RNA interference therapy in lung transplant patients infected with respiratory syncytial virus. Am J Respir Crit Care Med 183:531–538

    Article  Google Scholar 

  • Zhao Q-Q et al (2009) N/P ratio significantly influences the transfection efficiency and cytotoxicity of a polyethylenimine/chitosan/DNA complex. Biol Pharm Bull 32(4):706–710

    Article  Google Scholar 

Download references

Acknowledgments

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.

Author information

Author notes

    Authors and Affiliations

    1. Department of Pharmaceutical Technology, Faculty of Pharmacy and Biochemistry, University of Buenos Aires, Junín 954, 1113AAD, Buenos Aires, Argentina

      Sebastián Ezequiel Pérez & Adriana Mónica Carlucci

    2. Institute and Department of Biological Chemistry, Faculty of Pharmacy and Biochemistry, University of Buenos Aires, Junín 954, 1113AAD, Buenos Aires, Argentina

      Yamila Gándola & Lorena González

    Authors
    1. Sebastián Ezequiel Pérez
      View author publications

      You can also search for this author in PubMed Google Scholar

    2. Yamila Gándola
      View author publications

      You can also search for this author in PubMed Google Scholar

    3. Adriana Mónica Carlucci
      View author publications

      You can also search for this author in PubMed Google Scholar

    4. Lorena González
      View author publications

      You can also search for this author in PubMed Google Scholar

    Corresponding author

    Correspondence to Sebastián Ezequiel Pérez.

    Additional information

    Sebastián Ezequiel Pérez and Yamila Gándola contributed equally to this paper.

    Rights and permissions

    Reprints and permissions

    About this article

    Check for updates. Verify currency and authenticity via CrossMark

    Cite this article

    Pérez, S.E., Gándola, Y., Carlucci, A.M. et al. Development, characterization, and in vitro evaluation of phosphatidylcholine–sodium cholate-based nanoparticles for siRNA delivery to MCF-7 human breast cancer cells. J Nanopart Res 17, 128 (2015). https://doi.org/10.1007/s11051-015-2937-1

    Download citation

    • Received:

    • Accepted:

    • Published:

    • DOI: https://doi.org/10.1007/s11051-015-2937-1

    Keywords

    Search

    Navigation