A comparative study on generating hydroxyl radicals by single and two-frequency ultrasound with gold nanoparticles...

Sonodynamic therapy (SDT) is a new manner of killing cancer cells based on the cytotoxic interactions of ultrasound with sonosensitizing agents. It is shown that gold nanoparticles (GNPs) increase the efficiency of cavitation activity of ultrasound. In this study the influence of a single and/or two frequencies of ultrasound waves to generate hydroxyl radicals (^·OH) was assessed in the presence of protoporphyrin IX (PpIX) and/or GNPs. Ultrasound cavitation activity was determined by recording fluorescence signals from chemical terephthalic acid (TA) dosimeters with or without PpIX and/or GNPs at the frequencies of 0.8 and 2.4 MHz individually and aggregately. To study hydroxyl radicals, experiments were performed with and without hydroxyl radical scavengers mannitol, histidine, and sodium azide. Cavitation activity was amplified by increasing ultrasound intensity and exposure time. The cavitation activity induced by dual ultrasound frequency was remarkably higher than the summation of effects produced by individual frequencies. All three scavengers reduced the fluorescence signal level. The effect of GNPs on intensifying cavitation activity at higher frequency was greater than that of lower frequency. PpIX showed a more effective sonosensitizing property at the lower frequency. Also, estimated synergism at dual frequency irradiation was improved in the presence of GNPs. We found that GNPs increased hydroxyl radical production at 2.4 MHz and that PpIX increased hydroxyl radical production at 0.8 MHz. Dual frequency exposure was more effective than single frequency exposure. PpIX at low frequency and gold nanoparticles at high frequency both enhance sonodynamic treatment efficacy.