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Contributions of mechanical and sonochemical effects to cell membrane damage induced by single-shot pulsed ultrasound with adjacent microbubbles

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Abstract

Purpose

The objective was to investigate the contributions of mechanical effects due to kinetic force induced by the dynamic behavior of microbubbles and sonochemical effects due to free radicals produced by inertial cavitation to cell membrane damage under sonoporation conditions in which cells with adjacent microbubbles were irradiated with single-shot pulsed ultrasound.

Methods

The free radical scavenger cysteamine was used to control the occurrence of sonochemical effects, and the ratios of cells with membrane damage to intact cells were compared in the presence and absence of cysteamine. To determine the optimal dose of cysteamine, free radical production on exposure to burst pulse ultrasound was investigated using KI-starch solutions with different concentrations (0–5 mM) of cysteamine. High-speed observation of the dynamic behavior of Levovist microbubbles during ultrasound exposure was also carried out in the presence and absence of cysteamine, and the difference in the ratios of the maximum bubble diameter to the initial diameter was evaluated. Next, human prostate cancer cells with adjacent Levovist microbubbles were exposed to single-shot pulsed ultrasound with a center frequency of 1 MHz, a peak negative pressure of 1.1 MPa, and a pulse width of 3 μs, and the percentages of cells with membrane damage were evaluated by fluorescent microscopy using propidium iodide in the presence and absence of cysteamine.

Results

It was confirmed that cysteamine at a concentration of 5 mM completely suppressed sonochemical effects without causing a change in the dynamic response of microbubbles to pulsed ultrasound. The percentages of cells with membrane damage in the presence and absence of cysteamine (5 mM) were 10.3% ± 4.1% (n = 13) and 8.7% ± 3.9% (n = 9), respectively. No significant difference was found (P = 0.36).

Conclusion

The results indicate that cell membrane damage induced by single-shot pulsed ultrasound with adjacent microbubbles was due mainly to mechanical effects, not to sonochemical effects.

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References

  1. Newman CM, Lawrie A, Brisken AF, et al. Ultrasound gene therapy: on the road from concept to reality. Echocardiography 2001;18:339–447.

    Article  PubMed  CAS  Google Scholar 

  2. Miller DL, Pislaru SV, Greenleaf JE. Sonoporation: mechanical DNA delivery by ultrasonic cavitation. Somat Cell Mol Genet 2002;27:115–134.

    Article  PubMed  CAS  Google Scholar 

  3. Niidome T, Huang L. Gene therapy progress and prospects: nonviral vectors. Gene Ther 2002;9:1647–1652.

    Article  PubMed  CAS  Google Scholar 

  4. Mehier-Humbert S, Guy RH. Physical methods for gene transfer: improving the kinetics of gene delivery into cells. Adv Drug Deliv Rev 2005;57:733–753.

    Article  PubMed  CAS  Google Scholar 

  5. Bao S, Thrall BD. Transfection of a reporter plasmid into cultured cells by sonoporation in vitro. Ultrasound Med Biol 1997;23: 953–959.

    Article  PubMed  CAS  Google Scholar 

  6. Pislaru SV, Pislaru C, Kinnick RR, et al. Optimization of ultrasound-mediated gene transfer: comparison of contrast agents and ultrasound modalities. Eur Heart J 2003;24:1690–1698.

    Article  PubMed  CAS  Google Scholar 

  7. Miller DL, Quddus J. Lysis and sonoporation of epidermoid and phagocytic monolayer cells by diagnostic ultrasound activation of contrast agent gas bodies. Ultrasound Med Biol 2001;27:1107–1113.

    Article  PubMed  CAS  Google Scholar 

  8. Miller DL, Quddus J. Diagnostic ultrasound-induced membrane damage in phagocytic cells loaded with contrast agent and its relation to Doppler-mode images. IEEE Trans Ultrason Ferroelectr Freq Control 2002;49:1094–102.

    Article  PubMed  Google Scholar 

  9. Kudo N, Miyaoka T, Okada K, et al. Study on mechanism of cell damage caused by microbubbles exposed to ultrasound. In: Proceedings of 2002 IEEE Ultrasonics Symposium, vol 2. Munich: IEEE; 2002. p. 1383–1386.

    Google Scholar 

  10. Marmottant P, Hilgenfeldt S. Controlled vesicle deformation and lysis by single oscillating bubbles. Nature 2003;423:153–156.

    Article  PubMed  CAS  Google Scholar 

  11. Postema M, van Wamel A, Lancée CT, et al. Ultrasound-induced encapsulated microbubble phenomena. Ultrasound Med Biol 2004;30:827–840.

    Article  PubMed  Google Scholar 

  12. Riesz P, Kondo T. Free radical formation induced by ultrasound and its biological implications. Free Radic Biol Med 1992;13: 247–270.

    Article  PubMed  CAS  Google Scholar 

  13. Korosoglou G, Hardt SE, Bekeredjian R, et al. Ultrasound exposure can increase the membrane permeability of human neutrophil granulocytes containing microbubbles without causing complete cell destruction. Ultrasound Med Biol 2006;32:297–303.

    Article  PubMed  Google Scholar 

  14. Flynn HG. Generation of transient cavities in liquids by microsecond pulses of ultrasound. J Acoust Soc Am 1982;72:1926–1932.

    Article  Google Scholar 

  15. Crum LA, Fowlkes JB. Acoustic cavitation generated by microsecond pulses of ultrasound. Nature 1986;319:52–54.

    Article  Google Scholar 

  16. Fowlkes JB, Crum LA. Cavitation threshold measurements for microsecond length pulses of ultrasound. J Acoust Soc Am 1988;83:2190–2201.

    Article  PubMed  CAS  Google Scholar 

  17. Kondo T, Nishimura J, Kitagawa H, et al. Optimization of enhancement of therapeutic efficacy of ultrasound frequency-dependent effects on iodine formation from KI-starch solutions and ultrasound-induced killing of rat thymocytes. J Med Ultrasonics 2003; 30:93–101.

    Article  Google Scholar 

  18. Okada K, Kudo N, Niwa K, et al. A basic study on sonoporation with microbubbles exposed to pulsed ultrasound. J Med Ultrasonics 2005;32:3–11.

    Article  Google Scholar 

  19. Kondo T, Yoshii G. Effect of intensity of 1.2 MHz ultrasound on change in DNA synthesis of irradiated mouse L cells. Ultrasound Med Biol 1985;11:113–119.

    Article  PubMed  CAS  Google Scholar 

  20. Miller DL, Williams AR. Bubble cycling as the explanation of the promotion of ultrasonic cavitation in a rotating tube exposure system. Ultrasound Med Biol 1989;15:641–648.

    Article  PubMed  CAS  Google Scholar 

  21. Roots R, Okada S. Estimation of lifetime and diffusion distances of radicals involved in X-ray-induced DNA strand breaks or killing of mammalian cells. Radiat Res 1975;64:306–320.

    Article  PubMed  CAS  Google Scholar 

  22. Lawrie A, Brisken AF, Francis SE, et al. Ultrasound-enhanced transgene expression in vascular cells is not dependent upon cavitation-induced free radicals. Ultrasound Med Biol 2003;29: 1453–1461.

    Article  PubMed  Google Scholar 

  23. Clarke PR, Hill CR. Physical and chemical aspects of ultrasonic disruption of cells. J Acoust Soc Am 1970;47:649–653.

    Article  PubMed  CAS  Google Scholar 

  24. Young FR. Sonoluminescence from water containing dissolved gases. J Acoust Soc Am 1976;60:100–104.

    Article  CAS  Google Scholar 

  25. Kondo T, Misík V, Riesz P. Effect of gas-containing microspheres and echo contrast agents on free radical formation by ultrasound. Free Radic Biol Med 1998;25:605–612.

    Article  PubMed  CAS  Google Scholar 

  26. Edmonds PD, Sancier KM. Evidence for free radical production by ultrasonic cavitation in biological media. Ultrasound Med Biol 1983;9:635–639.

    Article  PubMed  CAS  Google Scholar 

  27. Unger EC, Porter T, Culp W, et al. Therapeutic applications of lipid-coated microbubbles. Adv Drug Deliv Rev 2004;56:1291–1314.

    Article  PubMed  CAS  Google Scholar 

  28. Lum AF, Borden MA, Dayton PA, et al. Ultrasound radiation force enables targeted deposition of model drug carriers loaded on microbubbles. J Control Release 2006;111:128–134.

    Article  PubMed  CAS  Google Scholar 

  29. Suzuki R, Takizawa T, Negishi Y, et al. Gene delivery by combination of novel liposomal bubbles with perfluoropropane and ultrasound. J Control Release 2007;117:130–136.

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Graduate School of Information Science and Technology, Hokkaido University, N14 W9, Sapporo, 060-0814, Japan

    Kengo Okada, Nobuki Kudo & Katsuyuki Yamamoto

  2. Department of Radiological Sciences, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Toyama, Japan

    Takashi Kondo

Authors
  1. Kengo Okada
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  2. Nobuki Kudo
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  3. Takashi Kondo
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  4. Katsuyuki Yamamoto
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Correspondence to Kengo Okada.

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Okada, K., Kudo, N., Kondo, T. et al. Contributions of mechanical and sonochemical effects to cell membrane damage induced by single-shot pulsed ultrasound with adjacent microbubbles. J Med Ultrasonics 35, 169–176 (2008). https://doi.org/10.1007/s10396-008-0192-0

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  • DOI: https://doi.org/10.1007/s10396-008-0192-0

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