Precision Visualization of Surgical Margins and Depth Imaging Performance in Orthotopic Hepatocellular Carcinoma Using Galactose-Targeted NIR-II Fluorescence Nanoprobe

Accurate identification of tumor margins during liver cancer resection is crucial for improving prognosis and reducing postoperative complications. This study investigated the feasibility of using a galactose-targeted nanofluorescent probe known as Gal-OH-BDP NPs (GOB NPs), which emits in the NIR-II window, for fluorescence imaging of orthotopic hepatocellular carcinoma in animal models. In vitro imaging was performed with Intralipid emulsion and chicken breast to assess the tissue penetration depth of the GOB NPs and ICG in the NIR window. In vivo NIR-I/II imaging was conducted in orthotopic liver tumor animal models. Pathological validation confirmed that intratumoral injection of GOB NPs resulted in targeted accumulation within liver cancer tissue. Quantitative image analysis revealed that GOB NPs-mediated NIR-II imaging exhibited the highest fluorescence intensity at the tumor site and the lowest background fluorescence in the liver, thus resulting in the highest tumor-to-normal liver tissue ratio (TNR) and the highest tumor margin resolution. Additionally, GOB NPs-mediated NIR-II imaging maintained effective tumor margin visibility at a depth of 8 mm, whereas ICG-mediated NIR-I/II imaging demonstrated effective detection depths limited to 2 and 5 mm, respectively. Following intravenous injection, the GOB NPs exhibited negative staining, thus clearly delineating the tumor boundaries and achieving a high contrast between the tumor and liver tissues.