Monitoring Muscle Perfusion in Rodents During Short-Term Ischemia Using Power Doppler Ultrasound

The aim of this work was to evaluate the reliability of power Doppler ultrasound (PD-US) measurements made without contrast enhancement to monitor temporal changes in peripheral blood perfusion. On the basis of pre-clinical rodent studies, we found that combinations of spatial registration and clutter filtering techniques applied to PD-US signals reproducibly tracked blood perfusion in skeletal muscle. Perfusion is monitored while modulating hindlimb blood flow. First, in invasive studies, PD-US measurements in deep muscle with laser speckle contrast imaging (LSCI) of superficial tissues made before, during and after short-term arterial clamping were compared. Then, in non-invasive studies, a pressure cuff was employed to generate longer-duration hindlimb ischemia. Here, B-mode imaging was also applied to measure flow-mediated dilation of the femoral artery while, simultaneously, PD-US was used to monitor downstream muscle perfusion to quantify reactive hyperemia. Measurements in adult male and female mice and rats, some with exercise conditioning, were included to explore biological variables. PD-US methods are validated through comparisons with LSCI measurements. As expected, no significant differences were found between sexes or fitness levels in flow-mediated dilation or reactive hyperemia estimates, although post-ischemic perfusion was enhanced with exercise conditioning, suggesting there could be differences between the hyperemic responses of conduit and resistive vessels. Overall, we found non-contrast PD-US imaging can reliably monitor relative spatiotemporal changes in muscle perfusion. This study supports the development of PD-US methods for monitoring perfusion changes in patients at risk for peripheral artery disease.