SignificanceShortwave-infrared (SWIR) imaging is reported to yield better contrast in fluorescence-guided surgery than near-infrared (NIR) imaging, due to a reduction in scattering. This benefit of SWIR was shown in animal studies, however not yet in clinical studies with patient samples.AimWe investigate the potential benefit of SWIR to NIR imaging in clinical samples containing cetuximab-IRDye800CW in fluorescence-guided surgery.ApproachThe potential of the epidermal growth factor-targeted NIR dye cetuximab-IRDye800CW in the shortwave range was examined by recording the absorption and emission spectrum. An ex vivo comparison of NIR and SWIR images using clinical tumor samples of patients with penile squamous cell carcinoma (PSCC) and head and neck squamous cell carcinoma (HNSCC) containing cetuximab-IRDye800CW was performed. The comparison was based on the tumor-to-background ratio and an adapted contrast-to-noise ratio (aCNR) using the standard of care pathology tissue assessment as the golden standard.ResultsBased on the emission spectrum, cetuximab-IRDye800CW can be detected in the SWIR range. In clinical PSCC samples, overall SWIR imaging was found to perform similarly to NIR imaging (NIR imaging is better than SWIR in the 2/7 criteria examined, and SWIR is better than NIR in the 3/7 criteria). However, when inspecting HNSCC data, NIR is better than SWIR in nearly all (5/7) examined criteria. This difference seems to originate from background autofluorescence overwhelming the off-peak SWIR fluorescence signal in HNSCC tissue.ConclusionSWIR imaging using the targeted tracer cetuximab-IRDye800CW currently does not provide additional benefit over NIR imaging in ex vivo clinical samples. Background fluorescence in the SWIR region, resulting in a higher background signal, limits SWIR imaging in HNSCC samples. However, SWIR shows potential in increasing the contrast of tumor borders in PSCC samples, as shown by a higher aCNR over a line.
Cancer continues to be a significant global health issue in the 21st century, presenting substantial risks to the health and well-being of individuals worldwide. Although there have been improvements in comprehending the molecular processes of the disease and creating treatments that specifically target it, a considerable portion of patients continue to encounter difficulties in attaining favorable results. Conventional two-dimensional (2D) cell cultures have been extensively used in cancer research. However, their inability to accurately mimic the intricate characteristics of tumors limits their effectiveness in predicting how anticancer treatments would perform in clinical settings. In order to overcome these restrictions, three-dimensional (3D) cell culture models, specifically multicellular spheroids, have arisen as promising tool for investigating cancer biology and therapeutic response. This work analyzes the development and growth dynamics of spheroids obtained from four distinct cancer cell lines: 9L-GFP, U251-RFP, A431, and FaDu. The stability and growth features of these spheroids were evaluated by culturing them using different cell counts and dilution ratios. Confocal microscopy was used to observe the formation of spheroids and measure their sizes for a duration of seven days. The results of our study reveal clear variations in growth patterns and stability profiles across the investigated cell lines. Notably, the 9L-GFP cell line demonstrates exceptional stability and continuous growth. The statistical analysis demonstrated that spheroids exhibited the most stability when the cell count was 25,000 cells and the dilution ratio was 1:3, as indicated by the high R-squared values. These findings highlight the significance of adjusting the number of cells and dilution ratios to ensure consistent and replicable spheroid formation. In summary, our study emphasizes the capacity of 3D spheroid models as effective instruments in cancer research and medication development, providing vital information about tumor biology and therapeutic responses in a context that closely resembles the physiological conditions.
In the United States, the annual incidence of oral squamous cell carcinoma (OSCC) exceeds 50,000 cases. Primary tumor resection remains the first line of treatment in these patients yet follow up neck dissection and chemoradiation treatment may be indicated if cancer has spread to tumor draining lymph nodes. There is a push to minimize morbidity from neck dissection by sentinel lymph node biopsy, where only the first lymph node(s) draining the primary tumor are excised and evaluated for cancer spread. However, with current pathology methods, results are not available to surgeons until patients have been sent home. In response, we are developing a method to rapidly stain and image whole excised lymph nodes in less than 30 min, so surgeons can react to positive cases while patients are still on the operating table. Here we present a human head and neck cancer spheroid model implanted in porcine lymph nodes as a means evaluating the potential for our staining and imaging protocols to rapidly identify cancer burden in lymph nodes.
An increasing number of cancer surgery protocols are including sentinel lymph node biopsies on the day of resection to stage for non-palpable spread of cancer through tumor draining lymph nodes. The challenge is that often a tumor-positive node will make it necessary to perform an enhanced resection of the lymphatic network, and if lymph node processing is not completed within the timeframe of surgery, then patients may have to be called back for additional surgery or have to undergo amplified chemo or radiation therapy. Our group is working on a rapid lymph node staining and fluorescence tomography system that we call ADEPT to provide surgeons with lymph node biopsy results within 15 min. The aim is to minimize the number of callback surgery or amplified therapy procedures to minimize stress to patients and reduce health care costs. This work predicts, using Monte Carlo photon propagation modeling simulations, that ADEPT has the potential to yield greater than 95% accuracy in detecting the smallest amount of cancer considered clinically relevant withing 15 min of tissue processing and imaging.
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