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Low-intensity ultrasound adjuvant therapy: enhancement of doxorubicin-induced cytotoxicity and the acoustic mechanisms involved

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Abstract

Purpose

In this study, the effects of low-intensity pulsed ultrasound (LIU) as an adjuvant to doxorubicin (DOX) treatment was further investigated in comparison to hyperthermia as another widely used adjuvant. The effects were compared with respect to cell killing and apoptosis induction in U937 cells. Human primary liver cancer (PLC) cells were also used to evaluate the effects of the combinations. The use of an echo contrast agent was investigated for further enhancement of cytotoxicity. Finally, the acoustic mechanisms involved were investigated.

Methods

The effects of different treatment regimens on cell viability were determined using the Trypan blue dye-exclusion test. Apoptosis induction was detected by flow cytometry using fluorescein isothiocyanate-annexin V and propidium iodide staining. The mechanistic study involved electron paramagnetic spin trapping for detecting free radical formation as an indicator of the occurrence of inertial cavitation and spectrophotometry for sucrose hydrolysis as an indicator for noncavitational effects.

Results

The combination treatments exerted synergistic effects on cytotoxicity depending on the acoustic conditions used. The use of LIU as an adjuvant to DOX treatment was shown to be superior to the use of hyperthermia as an adjuvant. Moreover, the combination seems to be promising for other cancer types provided that the acoustic conditions are properly selected with respect to drug concentration. The key ultrasound mechanism responsible for the synergism observed was shown to be the production of free radicals by inertial cavitation. Non-cavitational forces were also shown to contribute to the effect.

Conclusion

This study is motivating to engage in in vivo research with various cancer types as a step toward clinical applicability and is emphasizing on the importance of developing therapeutic protocols for setting LIU parameters with respect to other therapeutic conditions.

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Authors and Affiliations

  1. Department of Radiological Sciences, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, 2630 Sugitani, Toyama, 930-0194, Japan

    Takashi Kondo, Ryohei Ogawa, Mariame A. Hassan, Yukihiro Furusawa & Qing-Li Zhao

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

    Toru Yoshida & Kazuhiro Tsukada

  3. Department of Urology, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Toyama, Japan

    Akihiko Watanabe & Akihiro Morii

  4. Department of Anatomy, Fukuoka University School of Medicine, Fukuoka, Japan

    Loreto B. Feril Jr. & Katsuro Tachibana

  5. Laboratory of Animal Histophysiology, Division of Animal Science, Department of Bioresource Science, Graduate School of Agricultural Science, Kobe University, Kobe, Japan

    Hiroshi Kitagawa

  6. Life Science Research Center, University of Toyama, Toyama, Japan

    Yoshiaki Tabuchi & Ichiro Takasaki

  7. Department of Biochemistry, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Toyama, Japan

    Mohammad H. Shehata

  8. Laboratory of Biomedical Instrumentation and Measurements, Graduate School of Information Science and Technology, Hokkaido University, Sapporo, Japan

    Nobuki Kudo

Authors
  1. Takashi Kondo
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  2. Toru Yoshida
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  3. Ryohei Ogawa
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  4. Mariame A. Hassan
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  5. Yukihiro Furusawa
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  6. Qing-Li Zhao
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  7. Akihiko Watanabe
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  8. Akihiro Morii
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  9. Loreto B. Feril Jr.
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  10. Katsuro Tachibana
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  11. Hiroshi Kitagawa
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  12. Yoshiaki Tabuchi
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  13. Ichiro Takasaki
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  14. Mohammad H. Shehata
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  15. Nobuki Kudo
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  16. Kazuhiro Tsukada
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Corresponding author

Correspondence to Takashi Kondo.

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Kondo, T., Yoshida, T., Ogawa, R. et al. Low-intensity ultrasound adjuvant therapy: enhancement of doxorubicin-induced cytotoxicity and the acoustic mechanisms involved. J Med Ultrasonics 36, 61–68 (2009). https://doi.org/10.1007/s10396-009-0212-8

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  • DOI: https://doi.org/10.1007/s10396-009-0212-8

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