An in vitro study of the correlation between bubble distribution, acoustic emission, and cell damage by contrast...

The objective of this study was to investigate the influences of total exposure duration and pulse-to-pulse bubble distribution on contrast-mediated cell damage. Murine macrophage cells were grown as monolayers on thin polyester sheets. Contrast agent microbubbles were attached to these cells by incubation. Focused ultrasound exposures (P(r) = 2 MPa) were implemented at a frequency of 2.25 MHz with 46 cycle pulses and pulse repetition frequencies (PRF) of 1 kHz, 500 Hz, 100 Hz, and 10 Hz in a degassed water bath at 10 or 100 pulses. A 1 MHz receive transducer measured the scattered signal. The frequency spectrum was normalized to a control spectrum from linear scatterers. Photomicrographs were captured before, during, and after exposure at a frame rate of 2000 fps and a pixel resolution of 960 x 720. Results clearly show that cell death is increased, up to 60%, by increasing total exposure duration from 0 ms to 100 ms. There was an increasing difference in cell damage between a 10-pulse exposure and a 100-pulse exposure with increasing PRF. The greatest change in damage occurred at 1000 Hz PRF with a 53% increase between 10-pulse and 100-pulse exposures. For each pulse from 0 to 10, an overlay of the 2 mum bubble count with corresponding emission shows consistent behavior in its pulse-to-pulse changes, indicating a correlation between acoustic emission, bubble distribution, and cell damage.