Quantitative relations of acoustic inertial cavitation with sonoporation and cell viability

Ultrasound-induced acoustic cavitation assists gene delivery, possibly by increasing the permeability of the cell membranes. How the cavitation dose is related to the sonoporation rate and the cell viability is still unknown and so this in vitro study quantitatively investigated the effects of cavitation induced by 1-MHz pulsed ultrasound waves and the contrast agent Levovist® (containing microbubbles when reconstituted by adding saline and shaken) on the delivery of short DNA-FITC molecules into HeLa cells. The concentrations of cells and DNA-FITC were 2 × 105 cells/mL and 40 μg/mL, respectively. The cavitation was quantified as the inertial cavitation dose (ICD), corresponding to the spectral broadband signal enhancement during microbubble destruction. The relations of ICD with sonoporation and cell viability were examined for various acoustic pressures (0.48-1.32 MPa), Levovist® concentrations (1.12 × 105−1.12 × 107 bubbles/mL) and pulse durations (1-10 cycles). The linear regressions of the sonoporation rate versus ICD and the cell viability versus ICD were y = 28.67x + 10.71 (R2 = 0.95) and z = −62.83x + 91.18 (R2 = 0.84), respectively, where x is ICD, y is the sonoporation rate and z is the cell viability. These results show that the sonoporation rate and the cell viability are highly correlated with the ICD, indicating that sonoporation results may be potentially predicted using ICD. (E-mail: paichi@cc.ee.ntu.edu.tw)