Sonochemicals increase the mutation frequency of V79 cells in vitro

Phosphate buffered saline (PBS) was insonated or sham-insonated (1 MHz, 35 W/cm², continuous wave, 30 min) in rotating (200 rpm) sterile polystyrene culture tubes. After treatment, the PBS was used immediately to suspend washed Chinese hamster V79 cells in vitro. Cells were incubated in the PBS at 37°C for 15 min and then transferred to complete growth medium. Some insonation regimens also involved the inclusion of Albunex® (ALX; an ultrasound microbubble contrast agent) to enhance ultrasound-induced inertial cavitation. Following exposure to the pretreated PBS and 6 d of subculture in complete medium, the cells were assayed for plating efficiencies and mutation frequencies (resistance to 6-thioguanine). X-rays (3 Gy) served as a positive control. Cells exposed to insonated PBS with or without ALX or x-rays had statistically significantly elevated mean mutation frequencies (4.37 ± 0.97, 4.54 ± 1.00, and 24.28 ± 3.83 mutant colonies/10⁶ viable cells, respectively) relative to corresponding control regimens (ultrasound sham, 2.44 ± 0.56; x-ray sham, 2.96 ± 0.88 mutant colonies/10⁶ viable cells. The data supported the hypothesis that sonochemicals resulting from inertial cavitation have mutagenic potential.