Effects of low-intensity ultrasound on cell proliferation and reproductivity

Abstract

Ultrasound has been widely used in clinics. Cellular responses to low-intensity ultrasound are parameter-dependent. Proper parameter setting is vital to its exact use. To get guidelines for parameter setting, lowintensity ultrasound stimulation on the proliferation and reproductivity of HepG2 and 3T3 cells in vitro was examined with a 1.06 MHz-generator by changing the parameters(including intensity, pulse repetition frequency and duty cycle)in a wide range. Cell viability and reproductivity at different time after sonication were measured by 3- (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)and colony formation assay to indicate timerelated proliferation. The results illustrate that ultrasound irradiation at 0.4–0.8 W/cm2 and high pulse repetition frequency(100 Hz)can facilitate cell proliferation, while above 0.8 W/cm2 would resist it. The extent of resistance closely correlated with duty cycle and pulse repetition frequency. Resistance effect at low pulse repetition frequency(1 Hz)is greater than that at high pulse repetition frequency(100 Hz)and not time-related. The influence of high pulse repetition frequency is time-accumulated, indicating cellular process involved. These findings would provide valuable guidelines for the application of low-intensity ultrasound in stem cell transformation and tissue engineering.