Quantification of Kidney Inflammation Using Nanobubble-mediated Contrast Enhanced Ultrasound

Kidney inflammation is a central driver of acute kidney injury (AKI) and its progression to chronic kidney disease (CKD). While several imaging and biomarker-based approaches are under development, clinically validated non-invasive methods to directly quantify renal inflammation remain limited. This study introduces a novel approach using contrast-enhanced ultrasound (CEUS) with Cy5-labeled nanobubbles (NBs) to address this critical knowledge gap. Using a murine ischemia-reperfusion injury (IRI) model, CEUS imaging enabled real-time visualization of inflammation-induced changes in kidney perfusion and vascular integrity. Parametric analyses of non-linear imaging revealed delayed time-to-peak (TTP) and increased area under the falling curve (AUfC) in IRI kidneys, suggesting impaired microvascular perfusion and NB retention. Decorrelation time (DT) mapping further identified prolonged NB retention in the IRI group, indicating increased capillary permeability and NB extravasation. These findings correlated with histological and immunofluorescent analyses, which confirmed the presence of tubular injury, extravascular Cy5 signal localization, and increased neutrophil infiltration in inflamed kidney tissues. This study is the first to establish CEUS with NBs as a non-invasive, quantitative method for measuring kidney inflammation. With strong correlations between imaging metrics and histologic injury scores, this technology provides an accessible and non-invasive tool for monitoring renal inflammation and reducing reliance on invasive renal biopsies.