Ultrasound bioeffects in rats: quantification of cellular damage in the fetal liver after pulsed Doppler imaging

Abstract

To determine whether pulsed Doppler examination of the ductus venosus in rat fetuses could damage exposed tissue. On gestational day 18, the livers of a mean of approximately five fetuses per mother (n = 5.14, SD = 1.6), in a cohort of 35 pregnant female rats, were exposed individually to pulsed Doppler and these were considered the 'exposed group'. The remaining fetuses in each pregnant rat (n = 5.16, SD = 2.1) formed the 'control group'. We tested for 600, 300, 60, 20, 15, 10 and 3 s of exposure of the fetal ductus venosus and the damage was evaluated measuring a cell death index of apoptotic activity at 7 h post-exposure (n = 16). In addition, subgroups of mothers were sacrificed at 2, 4, 5, 7, 12 and 24 h post-exposure to determine when the damage appeared and disappeared and whether this depended on the exposure time. After exposure of 20 s or more, we observed significant damage, as assessed by caspase 3 activity (a marker of apoptotic activity related to tissue damage), in all cases; after 15 s of exposure, some samples presented damage (P = 0.4); there was no damage after 10 s or 3 s of exposure (P = 0.87 and P = 0.3, respectively). There was a positive linear correlation between apoptotic index and pulsed Doppler exposure time, (Pearson's coefficient = 0.324, P < 0.01). No liver still showed significant damage at 12 or 24 h post-exposure (P > 0.05 and P > 0.4). We observed reversible damage after pulsed Doppler imaging in an in-vivo fetal liver tissue rat model and found that longer exposure times produced more tissue damage. We established that 10 s was the maximum exposure time to ensure absence of damage to tissue in this model. It would appear sensible to recommend expert supervision of pulsed Doppler imaging and to have intervals between subsequent examinations.