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Background  Sonodynamic therapy (SDT) is a new approach for cancer treatment. Repair by reoxygenation induces cell damage in all treatment which uses photo- and sonosensitizers. In this study, the in vivo antitumor effect of dual-frequency sonication is investigated at low-level intensity and hematoporphyrin (Hp). It is used for the treatment of spontaneous breast adenocarcinoma of Balb/c mice with a variety of dose repetition and fractionation regimes.    Methods  Eighty tumor-bearing mice were divided into eight groups, the control group (A); the sham group (B); the injection of Hp alone group (C); 30-min dual-frequency sonication with Hp injection in one repetition at the first day group (D); two repetitions at the first and sixth days group (E); three repetitions at the first, sixth, and twelfth days group (F); four repetitions at the first, sixth, twelfth, and eighteenth days (30 min/repetition) group (G); and the fractional treatment group treated by dual-frequency sonication and Hp injection at the first, third, sixth, and ninth days (7.5 min/fraction) ( H ). For each group, the tumor growth delay was calculated during 30 days after treatment. These tumors were studied histopathologically.    Results  The results show that the treatment with ultrasound dose repetition in two, three, and four times (E, F, and G groups) were effective in delaying tumor growth compared with one-time sonication (D group) ( p  < 0.05). Also, the ultrasound dose fractionation is more effective in decreasing the tumor growth rate compared with the ultrasound dose repetition in four repeats and in one repeat from the 12th to the 30th day ( p  < 0.05). Histopathological studies indicated that the mitotic activity of tumor cells was reduced following treatment with four fraction and four repetition protocols.    Conclusion  The ultrasound dose fractionation and repetition technique with dual-frequency sonication can have a useful therapeutic effect in sonodynamic therapy with the possibility of use in future clinical applications.

Evaluation of fractionated and repeated sonodynamic therapy by using dual frequency for murine model of breast adenocarcinoma