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Molecular Pharmaceutics
Title Page Title: Development of a Nano-system for Targeted Eradication of Macrophage Ablation
Authors: Byeongtaek Oh 1 and Chi H. Lee, 1,* 1
Division of Pharmaceutical Sciences, School of Pharmacy, University of Missouri, Kansas City, MO 64108, USA *Corresponding Author Molecular Pharmaceutics
Chi H. Lee, Ph.D. Professor 2464 Charlotte Street, HSB-4242 Division of Pharmaceutical Sciences University of Missouri at Kansas City Kansas City, MO 64108 816-235 2408 (Tel) 816-235-5779 (Fax)
[email protected] ACS Paragon Plus Environment
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Molecular Pharmaceutics
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ABSTRACT This study was aimed to develop and evaluate a smart nano-system which targeted photo-thermal ablation of inflammatory macrophages in atherosclerotic plaque. Mannosylated-reduced graphene oxide (Man-rGO) was synthesized using three step procedures: (1) preparation of ox-GOs, (2) microwave-assisted synthesis of PEI-rGOs and (3) mannosylation of PEI-rGO using reductive amination reaction (Man-rGOs). The ζ-potential of Man-rGO that signifies electrophoretic mobility of the charged surface was examined using Zetasizer Nano ZS. The effects of Man-rGO on the cell viability was evaluated using LDH assay and MTS assay. The targeting efficacy of Man-rGO was assessed using the cellular uptake rate by M2-polarized (i.e., which is induced by IL-4) macrophage. The effects of NOMela loaded in Man-rGO on the enhancement of phagocytosis were evaluated by examining the phagocytic clearance rate of zymosan-FITC particles. The microwave-assisted reduction of GOs was adapted for a facile synthesis of polyethyleneimine-reduced GO (PEI-rGO). The mannose functionalization (Man-rGO) of PEI-rGO produced a greater number of amide linkages formed by reductive amination reaction between PEI-rGO and mannose. The ζ-potential of PEI-rGO was +30.6±3.3 mV, whereas that of Man-rGO was down to +13.1±3.8 mV upon interaction with mannose mainly due to the conjugation of mannose on the PEI-rGO surface. Near-infrared (NIR) irradiation increased the temperature of Man-rGO solution to around 45 °C, suggesting that Man-rGO is more potent than ox-GO or rGO in photothermal ablation activity triggered by NIR laser irradiation (808 nm). All testing formulations at the concentrations up to 10 µg/ml exerted less than 10% of membrane disintegration. For MTS study, more than 90% of cell viability were maintained at the concentrations (up to 10 µg/ml) of all tested formulations. The fluorescent microscopy images of cells after 1 hr incubation demonstrated that Man-rGO were mainly accumulated at the subcellular level where the mannose receptors were overexpressed. The cell viability of macrophages significant decreased upon exposure to Man-rGO irradiated with NIR, but no changes were observed from that of mast cells (for mast cells, 98.3±0.3%; for macrophages, 67.8±1.3%, p
