Quantitative photoacoustic imaging for early detection of muscle ischemia injury

Acute lower extremity ischemia is a limb-and life-threatening problem. The timing of clinical intervention is critical to achieving optimal outcomes. However, there has been a lack of effective techniques capable of evaluating muscle and limb damage. Microcirculatory injury is the initial pathological change during ischemic muscle injury. Here, we performed photoacoustic imaging (PAI) in real time to quantitatively detect the degree of microcirculatory injury of ischemic muscles in a rat model in which Evans blue (EB), which strongly binds to albumin in blood, was used as a nontoxic molecular PA probe. The right lower hind limbs of Sprague-Dawley (SD) rats were subjected to 2 or 3 hours of tourniquet-induced ischemia. Then, PA imaging of the tibialis anterior (TA) muscles in the anterior compartment was performed for 0-24 h after the release of compression. Twenty-four hours after reperfusion, rats were euthanized and examined for pathology, edema and muscle viability. Imaging at 680 nm on rats revealed that there was significant signal enhancement in the TA muscles of the two injury groups compared to the control group, and the 3-h injury group had significantly higher PA signal intensity than the 2-h injury group at each time point. Histopathology results obtained from both the normal and the damaged muscles correlated well with the PAI findings. In conclusion, PA imaging is a promising modality for quantitatively detecting limb and muscle ischemic injury and may pave the road for further clinical application.