title: Fluorescent molecular imaging of metastatic lymph node using near-infrared emitting low molecular weight heparin modified nanoliposome based on enzyme-substrate interaction.
abstract: Tumor metastatic lymph node mapping has been widely used to predict the metastatic spread of primary tumor and guide the lymph node dissection in clinical practice. In this research, a new near-infrared (NIR)-emitting low molecular weight heparin (LMWHEP)-modified Cy7-loaded nanoliposome (LMWHEP-NLips/Cy7) was developed and had the particle size of about 80 nm and the fluorescence intensity of about 2300, which is optimal for metastatic lymph node uptake and imaging. The NIR-emitting nanoliposomes were designed by LMWHEP coating on the surface of Cy7-loaded nanoliposome (NLips/Cy7) according to electrostatic attraction. The LMWHEP-NLips/Cy7 with negligible cytotoxicity for Hela and RAW264.7 cells and was found to be fluorescent stability compared with the Cy7-free dye at room temperature. The BALB/c nude mice bearing tumor lymphatic metastasis was established at eighth week post-injection by subcutaneously injecting Hela cells suspension. Heparanase (HPA) expression concentrations quantitatively measured by ELISA kit respectively were 237.42U/mL, 214.82U/mL and 128.45U/mL in the extracellular Hela cells, metastatic popliteal and iliac lymph node. LMWHEP-NLips/Cy7 successfully increased fluorescence signal in the metastatic lymph node compared with normal lymph node and achieve in vivo and ex vivo high fluorescence signal within 10 min and retention time up to 4 h post-injection. Maximum mean fluorescence intensity of the LMWHEP-NLips/Cy7 group was significantly more than NLips/Cy7 group (increase 2-fold in the metastatic popliteal lymph node and 4.8-fold in the metastatic iliac lymph node, p < 0.05). The experimental results demonstrating LMWHEP-NLips/Cy7 have the potential utility as specific, biosafe and stable near-infrared imaging contrast agents for HPA-expression tumor metastatic lymph node mapping. Copyright © 2016 John Wiley & Sons, Ltd.
authors: School of Pharmacy, Shenyang Pharmaceutical University, Shenyang, China.
authors: Tiantian
authors: T
authors: Ye
authors: School of Pharmacy, Shenyang Pharmaceutical University, Shenyang, China.
authors: Hefeng
authors: H
authors: Zhang
authors: Department of General Surgery, Peking Union Medical College hospital, Chinese Academy of Medical Sciences, Peking, China.
authors: Ge
authors: G
authors: Chen
authors: School of Pharmacy, China Medical University, Shenyang, China.
authors: Lei
authors: L
authors: Shang
authors: School of Pharmacy, Shenyang Pharmaceutical University, Shenyang, China.
authors: Shujun
authors: S
authors: Wang
mesh_terms: D000818:Animals; D003287:Contrast Media / Q000737:chemistry*; D005966:Glucuronidase / Q000032:analysis / Q000378:metabolism* / Q000494:pharmacology; D006367:HeLa Cells; D006495:Heparin, Low-Molecular-Weight / Q000528:radiation effects*; D064593:Heterografts; D006801:Humans; D007259:Infrared Rays; D008081:Liposomes; D008207:Lymphatic Metastasis / Q000000981:diagnostic imaging*; D051379:Mice; D057054:Molecular Imaging / Q000379:methods*; D053758:Nanoparticles / Q000737:chemistry; D061848:Optical Imaging / Q000379:methods*; D000067996:RAW 264.7 Cells; D000099175:Heparanase
chemical_list: D003287:Contrast Media; D005966:Glucuronidase; D006495:Heparin, Low-Molecular-Weight; D008081:Liposomes; D000099175:Heparanase
keywords: heparanase; low molecular weight heparin; lymph node mapping; metastasis; nanoliposome; near-infrared fluorescent