Fluorescence tomographic imaging of sentinel lymph node using near-infrared emitting bioreducible dextran nanogels

Jiejing Li,1* Beiqi Jiang,1* Chao Lin,2 Zhigang Zhuang1 1Department of Breast Surgery, Shanghai First Maternity and Infant Hospital, 2The Institute for Biomedical Engineering and Nanoscience, Tongji University School of Medicine, Tongji University, Shanghai, People’s Republic of China *These authors contributed equally to this work Abstract: Sentinel lymph node (SLN) mapping is a critical procedure for SLN biopsy and its diagnosis as tumor metastasis in clinical practice. However, SLN mapping agents used in the clinic frequently cause side effects and complications in the patients. Here, we report the development of a near-infrared (NIR) emitting polymeric nanogel with hydrodynamic diameter of ~28 nm – which is the optimal size for SLN uptake – for noninvasive fluorescence mapping of SLN in a mouse. This polymeric nanogel was obtained by coupling Cy7, an NIR dye, to the self-assembled nanogel from disulfide-linked dextran-deoxycholic acid conjugate with the dextran of 10 kDa, denoted as Dex–Cy7. Fluorescence imaging analysis showed that Dex–Cy7 nanogels had an enhanced photostability when compared to Cy7 alone. After intradermal injection of Dex–Cy7 nanogel into the front paw of a mouse, the nanogels were able to migrate into the mouse’s axillary lymph node, exhibiting longer retention time and higher fluorescence intensity in the node when compared to Cy7 alone. An immunohistofluorescence assay revealed that the nanogels were localized in the central region of lymph node and that the uptake was largely by the macrophages. In vitro and in vivo toxicity results indicated that the dextran-based nanogels were of low cytotoxicity at a polymer concentration up to 1,000 µg/mL and harmless to normal liver and kidney organs in mice at an intravenous dose of 1.25 mg/kg. The results of this study suggest that NIR-emitting polymeric nanogels based on bioreducible dextran-deoxycholic acid conjugates show high potential as fluorescence nanoprobes for safe and noninvasive SLN mapping. Keywords: nanogel, disulfide, dextran, lymph node, tomographic imagin