Ultrasound assessment of inflammation and renal tissue injury with microbubbles targeted to P-selectin

Abstract

Background - Routine methods capable of assessing tissue inflammation noninvasively are currently not available. We hypothesized that tissue retention of microbubbles targeted to the endothelial cell adhesion molecule P-selectin would provide a means to assess inflammation with ultrasound imaging. Methods and Results - Phospholipid microbubbles targeted to P-selectin (MBp) were created by conjugating monoclonal antibodies against murine P-selectin to the lipid shell. The microvascular behaviors of MBp and control microbubbles without antibody (MB) or with isotype control antibody (MBiso) were assessed by intravital microscopy of cremasteric venules of control and tumor necrosis factor (TNF)-α-stimulated wild-type mice. Retention of all microbubbles increased (P<0.05) with TNF-α treatment because of increased attachment to activated leukocytes. Extensive attachment of MBp directly to the venular endothelium or to adherent platelet-leukocyte aggregates was observed in TNF-α-stimulated mice, resulting in 4-fold greater (P<0.01) retention of MBp than either MBiso or MB. Enhanced retention of MBp was completely abolished in TNF-α-stimulated P-selectin-deficient mice. The ultrasound signal from microbubbles retained in inflamed tissue was assessed by contrast-enhanced renal ultrasound imaging of the kidneys of mice undergoing ischemia-reperfusion injury. In wild-type mice, this signal was significantly higher (P<0.05) for MBp (12±2 U) than either MBiso (6±3 U) or MB (5±3 U). In P-selectin-deficient mice, the signal for MBp was equivalent to that from control microbubbles. Conclusions - Microvascular retention of microbubbles targeted to P-selectin produces strong signal enhancement on ultrasound imaging of inflamed tissue. These results suggest that site-targeted microbubbles may be used to assess inflammation, tissue injury, and other endothelial responses noninvasively with ultrasound.