Journal of Nanoparticle Research

, Volume 10, Issue 3, pp 449–454 | Cite as

Factors that affect the efficiency of antisense oligodeoxyribonucleotide transfection by insonated gas-filled lipid microbubbles

Research Paper

Abstract

Objective: To investigate the factors that affect the efficiency of antisense oligodeoxyribonucleotide(AS-ODNs) transfection by insonated gas-filled lipid microbubbles. Methods: Lipid microbubbles filled with two types of gases–air and C3F8, were prepared respectively. An AS-ODNs sequence HA824 and a breast cancer cell line SK-BR-3 were used to define the various operating variables determining the transfection efficiency of insonated microbubbles. Two mixing methods, three levels of mixing speed, different mixing durations and various ultrasound initiation time after mixing were examined respectively. Transfection efficiency was detected by fluorescence microscopy. Results: C3F8 microbubbles gave higher levels of AS-ODNs transfection efficiency than air microbubbles in all test conditions. Transfection efficiency resulted from mixing method A (incubation of HA824 and microbubbles before mixing cells) did not show significant difference with that of mixing method B (without incubation of HA824 and microbubbles before mixing cells). Mixing speed, duration of mixing and ultrasound initiation time after mixing were central to determining HA824 transfection efficiency in vitro. The optimum parameters for SK-BR-3 cells were found at a mixing speed of 40–50 rpm for 30–60 s with less than 60 s delay before ultrasound. Conclusion: Ultrasound-mediated AS-ODNs transfection enhanced by C3F8–filled lipid microbubbles represents an effective avenue for AS-ODNs transfer.

Keywords

Antisense oligodeoxyribonucleotide Ultrasound Transfection Microbubbles Nanomedicine 

References

  1. Ambrosini E, Ceccherini-Silberstein F, Erfle V, Aloisi F, Levi G (1999) Gene transfer in astrocytes: comparison between different delivering methods and expression of the HIV-1 protein Nef. J Neurosci Res 55(5):569–577CrossRefGoogle Scholar
  2. Bell H, Kimber WL, Li M, Whittle IR (1998) Liposomal transfection efficiency and toxicity on glioma cell lines: in vitro and in vivo studies. Neuroreport 9(5):793–798CrossRefGoogle Scholar
  3. Christiansen JP, French BA, Klibanov AL, Kaul S, Lindner JR (2003) Targeted tissue transfection with ultrasound destruction of plasmid-bearing cationic microbubbles. Ultrasound Med Biol 29(12):1759–1767CrossRefGoogle Scholar
  4. Gogate PR, Wilhelm AM, Pandit AB (2003) Some aspects of the design of sonochemical reactors. Ultrason Sonochem 10(6):325–330CrossRefGoogle Scholar
  5. Greenleaf WJ, Bolander ME, Sarkar G, Goldring MB, Greenleaf JF (1998) Artificial cavitation nuclei significantly enhance acoustically induced cell transfection. Ultrasound Med Biol 24(4):587–595CrossRefGoogle Scholar
  6. Kim YJ, Kim TW, Chung H, Kwon IC, Sung HC, Jeong SY (2003) The effects of serum on the stability and the transfection activity of the cationic lipid emulsion with various oils. Int J Pharm 252(1–2):241–252CrossRefGoogle Scholar
  7. Lawrie A, Brisken AF, Francis SE, Wyllie D, Kiss-Toth E, Qwarnstrom EE, Dower SK, Crossman DC, Newman CM (2003) Ultrasound-enhanced transgene expression in vascular cells is not dependent upon cavitation-induced free radicals. Ultrasound Med Biol 29(10):1453–1461CrossRefGoogle Scholar
  8. Mack KD, Wei R, Elbagarri A, Abbey N, McGrath MS (1998) A novel method for DEAE-dextran mediated transfection of adherent primary cultured human macrophages. J Immunol Methods 211(1–2):79–86CrossRefGoogle Scholar
  9. Melkonyan H, Sorg C, Klempt M (1996) Electroporation efficiency in mammalian cells is increased by dimethyl sulfoxide (DMSO). Nucleic Acids Res 24(21):4356–4357CrossRefGoogle Scholar
  10. Miura S, Tachibana K, Okamoto T, Saku K (2002) In vitro transfer of antisense oligodeoxynucleotides into coronary endothelial cells by ultrasound. Biochem Biophys Res Commun 298(4):587–590CrossRefGoogle Scholar
  11. Nabel EG, Gordon D, Yang ZY, Xu L, San H, Plautz GE, Wu BY, Gao X, Huang L, Nabel GJ (1992) Gene transfer in vivo with DNA–liposome complexes: lack of autoimmunity and gonadal localization. Human Gene Ther 3(6):649–656CrossRefGoogle Scholar
  12. Porter TR, Iversen PL, Li S, Xie F (1996) Interaction of diagnostic ultrasound with synthetic oligonucleotide-labeled perfluorocarbon-exposed sonicated dextrose albumin microbubbles. J Ultrasound Med 15(8):577–584Google Scholar
  13. Price RJ, Kaul S (2002) Contrast ultrasound targeted drug and gene delivery: an update on a new therapeutic modality. J Cardiovasc Pharmascol Therapeut 7(3):171–180CrossRefGoogle Scholar
  14. Tzavelas C, Bildirici L, Rickwood D (2004) Factors that affect the efficiency of cell transfection by immunoporation. Anal Biochem 328(2):219–224CrossRefGoogle Scholar
  15. Unger E, Porter T, Culp W, Labell R, et al (2004) Therapeutic applications of lipid-coated microbubbles. Adv Drug Deliv Rev 56(9):1291–1314CrossRefGoogle Scholar
  16. Wang XH (2005) Gene transfer with Levovist and plasmid GFP into skeletal muscle cell. Chin J Ultrasonogr 14(6):464–466Google Scholar
  17. Watanabe Y, Nomoto H, Takezawa R, Miyoshi N, Akaike T (1994) Highly efficient transfection into primary cultured mouse hepatocytes by use of cation-liposomes: an application for immunization. J Biochem 116(6):1220–1226Google Scholar
  18. Zhao YZ, Luo YK, Zhang Y, Mei XG, Tang J (2005a) Property and contrast-enhancement effects of lipid ultrasound contrast agent: a preliminary experimental study. Ultrasound Med Biol 31(4):537–543CrossRefGoogle Scholar
  19. Zhao YZ, Liang HD, Mei XG, Halliwell M (2005b) Preparation, characterization and in vivo observation of phospholipid-based gas-filled microbubbles containing hirudin. Ultrasound in Med Biol 31(9):1237–1243CrossRefGoogle Scholar
  20. Zhao YZ, Luo YK, Tang J, Mei XG, Zhang Y, Lin Q (2006) Echogenic phospholipids-based gas-filled microbubbles as delivery system of antisense oligodeoxynucleotides. Yao Xue Xue Bao 41(9):899–904Google Scholar
  21. Zhong ZR, Zhang ZR, Liu J, Deng Y, Fu Y, Song QG, He Q (2007) Characterization of transferrin-modified procationic–liposome protamine-DNA complexes. Yakugaku Zasshi 127(3):533–539CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  1. 1.Department of Clinical PharmacologyGeneral Hospital of Beijing Military Command of PLABeijingChina
  2. 2.Madam Medical Management GroupBeijingChina

Personalised recommendations