Bioeffects caused by changes in acoustic cavitation bubble density and cell concentration: a unified explanation based on cell-to-bubble ratio and blast radius.

Acoustic cavitation has been shown to load drugs, proteins and DNA into viable cells as a complex function of acoustic and nonacoustic parameters. To better understand and quantify this functionality, DU145 prostate cancer cell suspensions at different cell concentrations (2.5 x 10(5) to 4.0 x 10(7) cells/mL) were exposed to 500 kHz ultrasound (US) over a range of acoustic energy exposures (2 to 817 J/cm(2); peak negative pressures of 0.64 to 2.96 MPa; exposure times of 120 to 2000 ms) in the presence of different initial concentrations of Optison contrast agent bubbles (3.6 x 10(4) to 9.3 x 10(7) bubbles/mL). As determined by flow cytometry, molecular uptake of calcein and cell viability both increased with increasing cell density; viability decreased and uptake was unaffected by increasing initial contrast agent concentration. When normalized relative to the initial contrast agent concentration (e.g., cells killed per bubble), bioeffects increased with increasing cell density and decreased with increasing bubble concentration. These varying effects of contrast agent concentration and cell density were unified through an overall correlation with cell-to-bubble ratio. Additional analysis led to estimation of "blast radii" over which bubbles killed or permeabilized cells; these radii were as much as 3 to 90 times the bubble radius. Combined, these results suggest that extensive molecular uptake into cells at high viability occurs for low-energy exposure US applied at a high cell-to-bubble ratio.