The effect of low frequency and low intensity ultrasound combined with microbubbles on the sonoporation efficiency of MDA-MB-231 cells

Ultrasound can produce certain biophysical effects including thermal and non-thermal effects on cells. Sonoporation, the most widely studied non-thermal biological effect of ultrasound, is considered to be the basis for new therapeutic applications. Ultrasound irradiation can increase the permeability of cell membranes through sonoporous effect, which makes molecules like those of drugs, protein, and DNA that normally cannot pass through the cell membranes be able to enter cells. Considering the poor therapeutic effect and poor prognosis of triple negative breast cancer, we aimed to explore the experimental conditions and find the optimal parameters to improve the therapeutic efficacy of chemotherapeutic drugs for MDA-MB-231 cells.

In our study, the optimal parameters of the sonoporous effect for MDA-MB-231 cells were 300 mW/cm2 of ultrasound intensity, 2 minutes of irradiation time, and 20% microbubbles concentration.

By establishing an experimental and control group, our study investigated the effect of low frequency and low intensity ultrasound combined with microbubbles on MDA-MB-231 cell membrane permeability at different times. We conducted factorial cross-design and set 3 levels of ultrasound intensity: 230, 300, and 370 mW/cm2; 3 levels of irradiation time: 1, 2, and 3 minutes; and 6 levels of microbubble doses: 0, 0.2, 0.4, 0.6, 0.8, and 1 mL.

Results show that ultrasound intensity, time of irradiation, and microbubbles concentration are not only related to but also have interactive effects on the sonoporation efficiency of MDA-MB-231 cells, with the rank order being sound intensity, irradiation time, and microbubble concentration. The average positive rates (%) of FD4 staining in sound intensities of 230, 300, and 370 mW/cm2 levels were 1.20±0.71, 13.80±5.86, and 10.71±4.36, respectively; and in irradiated times of 1, 2, and 3 min they were 7.54±5.95, 9.74±8.42, and 8.59±5.80, respectively. When the microbubbles increased according to the gradient of 0, 0.2, 0.4, 0.6, 0.8, and 1 mL, the positive rates (%) of FD4 staining were 7.32±5.89, 9.26±7.39, 8.31±5.67, 10.12±8.42, 8.67±7.23, and 7.72±6.24. Conclusions: In our study, the optimal parameters of the sonoporous effect for MDA-MB-231 cells were 300 mW/cm2 of ultrasound intensity, 2 minutes of irradiation time, and 20% microbubbles concentration.