Precise Detection of Surgical Margin in Head and Neck Cancer Using Dual Near-Infrared Imaging of the Tumor and Tumor Microenvironment

The precise determination of resection margins during head and neck cancer surgery remains an unmet clinical challenge, where balancing complete tumor removal with preservation of healthy tissue is critical. To address this, we developed a dual near-infrared (NIR) fluorescence imaging strategy targeting both tumor cells and the tumor microenvironment (TME) in head and neck squamous cell carcinoma (HNSCC). Armed with 2 small-molecule fluorophores, OCTL14 for tumor-specific imaging and cRGD-ZW800-PEG for TME visualization, we performed real-time intraoperative NIR imaging in a FaDu tongue cancer xenograft model. Fluorophores were administered intravenously, and their targeting efficiency was quantified via time-dependent tumor-to-background ratios (TBRs), with surgical margins validated by histopathology. Our results demonstrated robust detection of cancerous tissue (TBR > 2.0) and surrounding TME (TBR > 1.5) within 4 h post-injection. Histopathology confirmed OCTL14 uptake in tumor cells, while cRGD-ZW800-PEG localized to peritumoral regions and vasculature. This dual-imaging approach offers a promising tool for fluorescence-guided surgery, enabling precise margin delineation to reduce locoregional recurrence and perioperative complications, thereby improving patient outcomes and quality of life.