Dual-modality imaging with 99mTc and fluorescent indocyanine green using surface-modified silica nanoparticles for biopsy of the sentinel lymph node: an animal study

BACKGROUND: We propose a new approach to facilitate sentinel node biopsy examination by multimodality imaging in which radioactive and near-infrared (NIR) fluorescent nanoparticles depict deeply situated sentinel nodes and fluorescent nodes with anatomical resolution in the surgical field. For this purpose, we developed polyamidoamine (PAMAM)-coated silica nanoparticles loaded with technetium-99m ((99m)Tc) and indocyanine green (ICG). METHODS: We conducted animal studies to test the feasibility and utility of this dual-modality imaging probe. The mean diameter of the PAMAM-coated silica nanoparticles was 30 to 50 nm, as evaluated from the images of transmission electron microscopy and scanning electron microscopy. The combined labeling with (99m)Tc and ICG was verified by thin-layer chromatography before each experiment. A volume of 0.1 ml of the nanoparticle solution (7.4 MBq, except for one rat that was injected with 3.7 MBq, and 1 μg of an ICG derivative [ICG-sulfo-OSu]) was injected submucosally into the tongue of six male Wistar rats. RESULTS: Scintigraphic images showed increased accumulation of (99m)Tc in the neck of four of the six rats. Nineteen lymph nodes were identified in the dissected neck of the six rats, and a contact radiographic study showed three nodes with a marked increase in uptake and three nodes with a weak uptake. NIR fluorescence imaging provided real-time clear fluorescent images of the lymph nodes in the neck with anatomical resolution. Six lymph nodes showed weak (+) to strong (+++) fluorescence, whereas other lymph nodes showed no fluorescence. Nodes showing increased radioactivity coincided with the fluorescent nodes. The radioactivity of 15 excised lymph nodes from the four rats was assayed using a gamma well counter. Comparisons of the levels of radioactivity revealed a large difference between the high-fluorescence-intensity group (four lymph nodes; mean, 0.109% ± 0.067%) and the low- or no-fluorescence-intensity group (11 lymph nodes; mean, 0.001% ± 0.000%, p < 0.05). Transmission electron microscopy revealed that small black granules were localized to and dispersed within the cytoplasm of macrophages in the lymph nodes. CONCLUSION: Although further studies are needed to determine the appropriate dose of the dual-imaging nanoparticle probe for effective sensitivity and safety, the results of this animal study revealed a novel method for improved node detection by a dual-modality approach for sentinel lymph node biopsy

Comparison between squamous cell carcinoma and inflammatory diseases of the oral and maxillofacial region using gallium-67 scintigraphy with computed tomography and magnetic resonance imaging

Purpose: This study aimed to compare squamous cell carcinoma (SCC) with inflammatory diseases of the oral and maxillofacial region using gallium 67 (67Ga) scintigraphy with computed tomography (CT) and magnetic resonance imaging (MRI). Material and methods: Seventy patients with SCC and 15 patients with inflammatory diseases of the oral and maxillofacial region underwent 67Ga scintigraphy with CT and MRI. A comparison between imaging features of 67Ga scintigraphy, CT, and MRI and lesions was performed with the Pearson's chi-squared test. Results: 67Ga scintigraphy was positive for 25 of 39 patients with SCC positive on CT and/or MRI (64.1%) and for 13 of 15 patients with inflammatory diseases positive on CT and/or MRI (86.7%). The detection of inflammatory diseases with 67Ga scintigraphy was higher than that of SCC (p = 0.104). Conclusions: This study compared SCC with inflammatory diseases of the oral and maxillofacial region using 67Ga scintigraphy with CT and MRI. 67Ga scintigraphy is an effective technique for detection of inflammatory diseases of the oral and maxillofacial region

Structural variations in parotid glands induced by radiation therapy in patients with oral carcinoma observed on contrast-enhanced computed tomography

Background: Xerostomia is one of the commonest radiation-induced complications in patients with head and neck carcinoma. The aim of this study was to assess structural variations in parotid glands induced by radiation therapy in patients with oral carcinoma with contras-enhanced computed tomography (CECT). Material/Methods: A retrospective study was performed in 41 patients with oral carcinoma who underwent CECT for head and neck malignancies before and after radiotherapy. We analyzed the relationship between parotid density variations, parotid volume change, as seen on CECT, and the mean radiation dose applied to the parotid glands in patients with oral carcinoma immediately after radiotherapy, and 2 and 3 years later. Results: Immediately after radiotherapy, high-density changes on contrast-enhanced CT were observed in 70.5% of the irradiated parotids. Low-density changes due to fat degeneration were seen in 46.2% and 72.2% of the irradiated parotids 2 and 3 years after radiotherapy, respectively. The mean dose applied to the parotids with the low-density changes and without such changes 3 years after radiotherapy was 46.0 Gy and 27.7 Gy, respectively (p=0.049). Furthermore, parotid shrinkage was observed in 63.6% of the irradiated parotids. Conclusions: This study suggests that the structural variations in parotid glands induced by radiotherapy included high-density changes that were observed immediately after radiotherapy and low-density changes that were seen at late follow-up. This study should be useful for clinicians in the assessment of radiation-induced injuries in the parotids with respect to early prediction of xerostomia

Gallium-67 SPECT-CT for the evaluation of head and neck : preliminary study on maximum standardised uptake value in lesions, and in the parotid and submandibular glands

Purpose: The aim of this study was to investigate the gallium-67 (67Ga) single-photon emission-computed tomography (SPECT-CT) for the evaluation of head and neck, especially maximum standardised uptake value (SUVmax) in lesions, and in the parotid and submandibular glands. Material and methods: Fourteen patients with head and neck lesions who underwent SPECT-CT at 72 hours after injection of 67Ga were included in this study. The 67Ga SPECT-CT parameters SUVmax were compared between the parotid glands and submandibular glands. Statistical analyses for the SUVmax were performed by Mann-Whitney U test. A p value lower than 0.05 was considered as statistically significant. Results: Of all 14 patients, 67Ga SPECT-CT was positive for two cases (one malignant lymphoma and one osteomye­litis). The SUVmax for malignant lymphoma and osteomyelitis were 11.21 and 10.95, respectively. Furthermore, SUVmax for parotid glands and submandibular glands of all patients were 4.86 ± 1.89 and 4.64 ± 1.64, respectively (p = 0.694). Conclusions: This study investigated the 67Ga SPECT-CT for the evaluation of head and neck, especially SUVmax of lesions, and the parotid and submandibular glands. 67Ga SPECT-CT may be an effective technique for the evaluation of maxillofacial lesions

Dual-Labeled Near-Infrared/99mTc Imaging Probes Using PAMAM-Coated Silica Nanoparticles for the Imaging of HER2-Expressing Cancer Cells

We sought to develop dual-modality imaging probes using functionalized silica nanoparticles to target human epidermal growth factor receptor 2 (HER2)-overexpressing breast cancer cells and achieve efficient target imaging of HER2-expressing tumors. Polyamidoamine-based functionalized silica nanoparticles (PCSNs) for multimodal imaging were synthesized with near-infrared (NIR) fluorescence (indocyanine green (ICG)) and technetium-99m (99mTc) radioactivity. Anti-HER2 antibodies were bound to the labeled PCSNs. These dual-imaging probes were tested to image HER2-overexpressing breast carcinoma cells. In vivo imaging was also examined in breast tumor xenograft models in mice. SK-BR3 (HER2 positive) cells were imaged with stronger NIR fluorescent signals than that in MDA-MB231 (HER2 negative) cells. The increased radioactivity of the SK-BR3 cells was also confirmed by phosphor imaging. NIR images showed strong fluorescent signals in the SK-BR3 tumor model compared to muscle tissues and the MDA-MB231 tumor model. Automatic well counting results showed increased radioactivity in the SK-BR3 xenograft tumors. We developed functionalized silica nanoparticles loaded with 99mTc and ICG for the targeting and imaging of HER2-expressing cells. The dual-imaging probes efficiently imaged HER2-overexpressing cells. Although further studies are needed to produce efficient isotope labeling, the results suggest that the multifunctional silica nanoparticles are a promising vehicle for imaging specific components of the cell membrane in a dual-modality manner

HER2-Targeted Multifunctional Silica Nanoparticles Specifically Enhance the Radiosensitivity of HER2-Overexpressing Breast Cancer Cells

We investigated the effects of targeted functionalized silica nanoparticles on the radiosensitivity of cancer cells. Better control of the local concentration of silica nanoparticles may facilitate their use as an adjuvant in conjunction with ionizing radiation to target cancer cells while preventing damage to normal cells. Hyperbranched polyamidoamine (PAMAM) was grafted onto the surface of amorphous silica nanoparticles to functionalize them. The PAMAM-coated silica nanoparticles (PCSNs) were then conjugated with fluorescent dyes. Anti-HER2 antibodies were covalently attached to the labeled PCSNs. The HER2-overexpressing SK-BR3 breast cancer cell line was incubated in medium containing the PCSN probes. After incubation; the cells were exposed to X-ray radiation. Cells were counted in all samples using cell proliferation assays; and apoptotic cells were detected. The cell survival results showed that the combination of the targeted PCSN probes and radiation reduced the survival rate of SK-BR3 cells to a greater extent than when either PCSN probes, PCSNs or radiation were applied individually. The results also showed an increase in apoptosis in the SK-BR3 cells that internalized the PCSN probes and were then irradiated. Based on these data, PCSN probes act as specific radiosensitizing agents for HER2-overexpressing cells

Structural Variations in Parotid Glands Induced by Radiation Therapy in Patients with Oral Carcinoma Observed on Contrast-Enhanced Computed Tomography

Background: Xerostomia is one of the commonest radiation-induced complications in patients with head and neck carcinoma. The aim of this study was to assess structural variations in parotid glands induced by radiation therapy in patients with oral carcinoma with contras-enhanced computed tomography (CECT). Material/Methods: A retrospective study was performed in 41 patients with oral carcinoma who underwent CECT for head and neck malignancies before and after radiotherapy. We analyzed the relationship between parotid density variations, parotid volume change, as seen on CECT, and the mean radiation dose applied to the parotid glands in patients with oral carcinoma immediately after radiotherapy, and 2 and 3 years later. Results: Immediately after radiotherapy, high-density changes on contrast-enhanced CT were observed in 70.5% of the irradiated parotids. Low-density changes due to fat degeneration were seen in 46.2% and 72.2% of the irradiated parotids 2 and 3 years after radiotherapy, respectively. The mean dose applied to the parotids with the low-density changes and without such changes 3 years after radiotherapy was 46.0 Gy and 27.7 Gy, respectively (p=0.049). Furthermore, parotid shrinkage was observed in 63.6% of the irradiated parotids. Conclusions: This study suggests that the structural variations in parotid glands induced by radiotherapy included high-density changes that were observed immediately after radiotherapy and low-density changes that were seen at late follow-up. This study should be useful for clinicians in the assessment of radiation-induced injuries in the parotids with respect to early prediction of xerostomia

Imaging of Metastatic Cancer Cells in Sentinel Lymph Nodes using Affibody Probes and Possibility of a Theranostic Approach

The accurate detection of lymph node metastases is essential for treatment success in early-stage malignant cancer. Sentinel lymph node (SLN) biopsy is the most effective procedure for detecting small or micrometastases that are undetectable by conventional imaging modalities. To demonstrate a new approach for developing a more efficient SLN biopsy procedure, we reported a two-stage imaging method combining lymphoscintigraphy and near-infrared (NIR) fluorescence imaging to depict metastatic cancer cells in SLNs in vivo. Furthermore, the theranostic potential of the combined procedure was examined by cell culture and xenograft mouse model. Anti-HER2 and anti-epidermal growth factor receptor (EGFR) affibody probes were used for NIR fluorescence imaging. Strong NIR fluorescence signal intensity of the anti-EGFR affibody probe was observed in SAS cells (EGFR positive). Radioactivity in the SLNs was clearly observed in the in vivo studies. High anti-EGFR affibody NIR fluorescence intensity was observed in the metastatic lymph nodes in mice. The addition of the IR700-conjugated anti-EGFR affibody to the culture medium decreased the proliferation of SAS cells. Decreased proliferation was shown in Ki-67 immunohistochemistry in xenograft tumors. Our data suggest that a two-stage combined imaging method using lymphoscintigraphy and affibody probes may offer the direct visualization of metastatic lymph nodes as an easily applied technique in SLN biopsy. Although further animal studies are required to assess the effect of treating lymphatic metastasis in this approach, our study results provide a foundation for the further development of this promising imaging and treatment strategy for earlier lymph node metastasis detection and treatment