Advertisement

The Journal of Membrane Biology

, Volume 249, Issue 5, pp 677–689 | Cite as

Intracellular Delivery of Bleomycin by Combined Application of Electroporation and Sonoporation in Vitro

  • Mindaugas Tamošiūnas
  • Lluis M. Mir
  • Wen-Shiang Chen
  • Alexey Lihachev
  • Mindaugas Venslauskas
  • Saulius Šatkauskas
Article

Abstract

In this study, we aimed to determine whether the combination of electroporation (EP) and ultrasound (US) waves (sonoporation) can result in an increased intracellular delivery of anticancer drug bleomycin. CHO cells were treated with electric pulses (1 or 8 high voltage pulses of 800 or 1200 V/cm, 100 μs or 1 low voltage pulse of 100 or 250 V/cm, 100 ms) and with 880 kHz US of 320 or 500 kPa peak negative pressure, 100 % duty cycle, applied for 2 s in the presence or absence of exogenously added contrast agent microbubbles. Various sequential or simultaneous combinations of EP and sonoporation were used. The results of the study showed that i) sequential treatment of cells by EP and sonoporation enhanced bleomycin electrosonotransfer at the reduced energy of electric field and US; ii) sequential combination of EP and sonoporation induced a summation effect which at some conditions was more prominent when the cells were treated first by EP and then by sonoporation; iii) the most efficient intracellular delivery of bleomycin was achieved by the simultaneous application of cell EP and sonoporation resulting in percentage of reversibly porated cells above the summation level; and iv) compared with sequential application of EP and sonoporation, simultaneous use of electric pulses and US increased cell viability in the absence of bleomycin.

Keywords

Electroporation Sonoporation Drug delivery Electrosonoporation Ultrasound Microbubbles 

Notes

Acknowledgments

This work was funded by Research Council of Lithuania project TAP-03/2012 (scientific cooperation between Lithuania, Latvia and Taiwan), the project from National Science Council, Taiwan, NSC 100-2923-B-002-004-MY3. In addition, this work was partially supported by Culture Department of French Embassy in Lithuania, CNRS, Univ. Paris-Sud, Gustave Roussy, Fondation EDF and Agence Nationale de la Recherche through the Project Soundelivery ANR-14-CE17-0008. This work was also partly performed in the scope of the LEA EBAM, the European Associated laboratory Electric pulses in Biology and Medicine.

References

  1. Chen WS, Brayman AA, Matula TJ, Crum LA (2003) Inertial cavitation dose and hemolysis produced in vitro with or without Optison. Ultrasound Med Biol 29:725–737CrossRefPubMedGoogle Scholar
  2. Čorović S, Al Sakere B, Haddad V, Miklavčič D, Mir LM (2008) Importance of contact surface between electrodes and treated tissue in electrochemotherapy. TCRT 7:393–399Google Scholar
  3. Deng CX, Sieling F, Pan HA, Cui J (2004) Ultrasound-induced cell membrane porosity. Ultrasound Med Biol 30:519–526CrossRefPubMedGoogle Scholar
  4. Escoffre JM, Kaddur K, Rols MP, Bouakaz A (2010) In vitro gene transfer by electrosonoporation. Ultrasound Med Biol 36:1746–1755CrossRefPubMedGoogle Scholar
  5. Hallow DM, Mahajan AD, McCutchen TE, Prausnitz MR (2006) Measurement and correlation of acoustic cavitation with cellular bioeffects. Ultrasound Med Biol 32:1111–1122CrossRefPubMedGoogle Scholar
  6. Jakštys B, Ruzgys P, Tamošiūnas M, Šatkauskas S (2015) Different Cell Viability Assays Reveal Inconsistent Results After Bleomycin Electrotransfer In Vitro. J Membr Biol 248:857–863CrossRefPubMedGoogle Scholar
  7. Kotnik T, Macek-Lebar A, Miklavcic D, Mir LM (2000) Evaluation of cell membrane electropermeabilization by means of a nonpermeant cytotoxic agent. Biotechniques 28:921–926PubMedGoogle Scholar
  8. Kotopoulis S, Delalande A, Popa M, Mamaeva V, Dimcevski G, Gilja OH, Postema M, Gjertsen BT, McCormack E (2014) Sonoporation-enhanced chemotherapy significantly reduces primary tumour burden in an orthotopic pancreatic cancer xenograft. Mol Imaging Biol 16:53–62CrossRefPubMedGoogle Scholar
  9. Larkin J, Soden D, Collins C, Tangney M, Preston JM, Russell LJ, McHale AP, Dunne C, O’Sullivan GC (2005) Combined electric field and ultrasound therapy as a novel anti-tumour treatment. Eur J Cancer 41:1339–1348CrossRefPubMedGoogle Scholar
  10. Lee EW, Loh CT, Kee ST (2007) Imaging guided percutaneous irreversible electroporation: ultrasound and immunohistological correlation. TCRT 6:287–294Google Scholar
  11. Liu Y, Miyoshi H, Nakamura M (2006) Encapsulated ultrasound microbubbles: therapeutic application in drug/gene delivery. J Controlled Release 114:89–99CrossRefGoogle Scholar
  12. Longsine-Parker W, Wang H, Koo C, Kim J, Kim B, Jayaraman A, Han A (2013) Microfluidic electro-sonoporation: a multi-modal cell poration methodology through simultaneous application of electric field and ultrasonic wave. Lab Chip 13:2144–2152CrossRefPubMedGoogle Scholar
  13. Marmottant P, Hilgenfeldt S (2003) Controlled vesicle deformation and lysis by single oscillating bubbles. Nature 423:153–156CrossRefPubMedGoogle Scholar
  14. Mehier-Humbert S, Bettinger T, Yan F, Guy RH (2005) Plasma membrane poration induced by ultrasound exposure: implication for drug delivery. J Control Release 104:213–222CrossRefPubMedGoogle Scholar
  15. Miller DL, Quddus J (2000) Sonoporation of monolayer cells by diagnostic ultrasound activation of contrast agent gas bodies. Ultrasound Med Biol 26:661–667CrossRefPubMedGoogle Scholar
  16. Mir LM, Orlowski S, Belehradek J Jr, Paoletti C (1991) Electrochemotherapy potentiation of antitumour effect of bleomycin by local electric pulses. Eur J Cancer 27:68–72CrossRefPubMedGoogle Scholar
  17. Prentice P, Cuschierp A, Dholakia K, Prausnitz M, Campbell P (2005) Membrane disruption by optically controlled microbubble cavitation. Nat Phys 1:107–110CrossRefGoogle Scholar
  18. Rollan-Haro AM, Smyth A, Hughes P, Reid CN, McHale AP (2005) Electro-sensitisation of mammalian cells and tissues to ultrasound: a novel tumour treatment modality. Cancer Lett 222:49–55CrossRefPubMedGoogle Scholar
  19. Tamošiūnas M, Jurkonis R, Mir LM, Lukoševičius A, Venslauskas MS, Šatkauskas S (2012a) Microbubble sonodestruction rate as a metric to evaluate sonporation efficiency. J Ultrasound Med 31:1993–2000PubMedGoogle Scholar
  20. Tamošiūnas M, Jurkonis R, Mir LM, Lukoševičius A, Venslauskas MS, Šatkauskas S (2012b) Adjustment of ultrasound exposure duration to microbubble sonodestruction kinetics for optimal cell sonoporation in vitro. TCRT 11:375–387Google Scholar
  21. Tomizawa M, Ebara M, Saisho H, Sakiyama S, Tagawa M (2001) Irradiation with ultrasound of low output intensity increased chemosensitivity of subcutaneous solid tumors to an anti-cancer agent. Cancer Lett 173:31–35CrossRefPubMedGoogle Scholar
  22. van Wamel A, Kooiman K, Harteveld M, Emmer M, ten Cate F, Versluis M, de Jong N (2006) Vibrating microbubbles poking individual cells: drug transfer into cells via sonoporation. J. Control Release 112:149–155CrossRefPubMedGoogle Scholar
  23. Venslauskas MS, Šatkauskas S, Rodaitė-Riševičienė R (2010) Efficiency of the delivery of small charged molecules into cells in vitro. Bioelectrochemistry 79:130–135CrossRefPubMedGoogle Scholar
  24. Ward M, Wu J, Chiu JF (1999) Ultrasound induced cell lysis and sonoporation enhanced by contrast agents. J Acoust Soc Am 105:2951–2957CrossRefPubMedGoogle Scholar
  25. Weaver JC, Chizmadzhev YuA (1996) Theory of electroporation: a review. Bioelectrochem Bioenerg 41:135–160CrossRefGoogle Scholar
  26. Yamashita YI, Shimada M, Tachibana K, Harimoto N, Tsujita E, Shirabe K, Miyazaki JI, Sugimachi K (2002) In vivo gene transfer into muscle via electro-sonporation. Hum Gene Ther 13:2079–2084CrossRefPubMedGoogle Scholar
  27. Yamashita YI, Shimada M, Minagawa R, Tsujita E, Harimoto N, Tanaka S, Shirabe K, Miyazaki JI, Maehara Y (2004) Muscle-targeted interleukin-12 gene therapy of orthotopic hepatocellular carcinoma in mice using in vivo electrosonoporation. Mol Cancer Ther 3:177–182Google Scholar
  28. Zhou Y, Kumon RE, Cui J, Deng CX (2009) The size of sonoporation pores on the cell membrane. Ultrasound Med Biol 35:1756–1760CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Mindaugas Tamošiūnas
    • 1
  • Lluis M. Mir
    • 2
  • Wen-Shiang Chen
    • 3
  • Alexey Lihachev
    • 4
  • Mindaugas Venslauskas
    • 1
  • Saulius Šatkauskas
    • 1
  1. 1.Biophysical research group, Faculty of Natural SciencesVytautas Magnus UniversityKaunasLithuania
  2. 2.Vectorology and Anticancer Therapeutics, UMR 8203, Univ. Paris-Sud Gustave RoussyUniversité Paris-SaclayVillejuifFrance
  3. 3.Department of Physical Medicine & RehabilitationNational Taiwan University HospitalTaipeiTaiwan
  4. 4.Institute of Atomic Physics and SpectroscopyUniversity of LatviaRīgaLatvia

Personalised recommendations