In vivo visualization and characterization of inflamed intestinal wall: the exploration of targeted microbubbles in assessing NF‐κB expression

NF‐κB, a critical cytokine of inflammatory bowel diseases (IBD), is a viable marker to reflect the inflammatory activity of the intestine. We aimed to develop NF‐κB‐targeted microbubbles (MBs) and perform molecular contrast‐enhanced ultrasound (CEUS) to quantify NF‐κB expressions on the intestinal wall in IBD mice in vivo. In this study, NF‐κB‐targeted MBs were fabricated by connecting biotin‐loaded NF‐κB antibodies and avidin‐loaded MBs. NF‐κB‐targeted MBs presented as transparent and round bubbles with an average diameter of 1.03/μm±0.01. The specific binding of targeted MBs and inflammatory cells was validated by in vitro experiments, including flow cytometry, Western blot and immunofluorescence, which revealed the specific binding of targeted MBs and inflammatory cells. Subsequently, NF‐κB‐targeted CEUS imaging was performed on mice with chemical‐induced colitis, and the peak intensity (PI) and time‐to‐peak (TTP) were quantified. Pathological and immunohistochemical (IHC) examinations were further implemented. For the target CEUS group, fast enhancement followed by slow subsiding was observed. The PI of target CEUS of the IBD mice was significantly higher than that of non‐target CEUS of the IBD mice, healthy controls and target CEUS of the treated IBD mice (34835%[13379–73492%] VS 437%[236–901%], 130%[79–231%], 528%[274–779%], p<0.0001), in accordance with the IHC results of NF‐κB expressions. The TTP of target CEUS of the treated mice was significantly higher than that of untreated mice (35.7s [18.1–49.5s] VS 8.3s [4.2–12.5s], p<0.0001). Therefore, we suggested that NF‐κB‐targeted CEUS could accurately detect and quantify NF‐κB expressions on the intestinal walls of IBD, enabling the evaluation of intestinal inflammation.