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WIREs Nanomed Nanobiotechnol
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Graphene‐based nanomaterials as molecular imaging agents

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Molecular imaging (MI) is a noninvasive, real‐time visualization of biochemical events at the cellular and molecular level within tissues, living cells, and/or intact objects that can be advantageously applied in the areas of diagnostics, therapeutics, drug discovery, and development in understanding the nanoscale reactions including enzymatic conversions and protein–protein interactions. Consequently, over the years, great advancement has been made in the development of a variety of MI agents such as peptides, aptamers, antibodies, and various nanomaterials (NMs) including single‐walled carbon nanotubes. Recently, graphene, a material popularized by Geim & Novoselov, has ignited considerable research efforts to rationally design and execute a wide range of graphene‐based NMs making them an attractive platform for developing highly sensitive MI agents. Owing to their exceptional physicochemical and biological properties combined with desirable surface engineering, graphene‐based NMs offer stable and tunable visible emission, small hydrodynamic size, low toxicity, and high biocompatibility and thus have been explored for in vitro and in vivo imaging applications as a promising alternative of traditional imaging agents. This review begins by describing the intrinsic properties of graphene and the key MI modalities. After which, we provide an overview on the recent advances in the design and development as well as physicochemical properties of the different classes of graphene‐based NMs (graphene‐dye conjugates, graphene‐antibody conjugates, graphene‐nanoparticle composites, and graphene quantum dots) being used as MI agents for potential applications including theranostics. Finally, the major challenges and future directions in the field will be discussed. WIREs Nanomed Nanobiotechnol 2015, 7:737–758. doi: 10.1002/wnan.1342 This article is categorized under: Diagnostic Tools > In Vivo Nanodiagnostics and Imaging Nanotechnology Approaches to Biology > Nanoscale Systems in Biology
A schematic of the molecular imaging modalities in the realm of graphene‐based nanomaterials.
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In vivo experimental data. (a) Relative tumor volumes of mice (n = 6) treated with DGO, GQD, DGO‐Cur, GQD‐Cur, Cur, and control (PBS); (b) Relative tumor weight of mice (n = 6) treated with DGO, GQD, DGO‐Cur, GQD‐Cur, Cur, and PBS; (c) Photographs of mice treated with DGO, GQD, DGO‐Cur, GQD‐Cur, Cur, and PBS after 14 days; (d) Photographs of tumors after 14 days of treatment with DGO, GQD, DGO‐Cur, GQD‐Cur, Cur, and PBS; (e) in vivo imaging of tumor‐bearing mice after injection of GQDs and GQD‐Cur (10 mg/kg). (Reprinted with permission from Ref . Copyright 2014 Nature Publishing Group)
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Schematic of the preparation of various curcumin‐graphene composites (GO‐Cur, DGO‐Cur, and GQD‐Cur) and their relative anticancer effects. (Reprinted with permission from Ref . Copyright 2014 Nature Publishing Group)
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Fluorescent microscopic images of HeLa cells directly labeled by graphene‐HQDs‐Trf‐1 for 4 h (a), and DOX‐graphene‐HQDs‐Trf‐1 for 6 h (b), 12 h (c), and 24 h (d). (i, fluorescent images by blue light excitation; ii, fluorescent images by green light excitation.) (Reprinted with permission from Ref . Copyright 2013 American Chemical Society)
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Preparation of water dispersible doxorubicin (DOX)‐graphene‐HQDs‐Trf. (a) Graphene oxide (GO). (b) PSS coated graphene. (c) Graphene‐HQDs. (d) Graphene‐HQDs‐Trf. (e) DOX‐graphene‐HQDs‐Trf. (Reprinted with permission from Ref . Copyright 2013 American Chemical Society)
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Chemical structures of different classes of hydrophilic polymers frequently used for functionalization of graphene subtypes in MI.
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Clinical MRI and X‐ray CT results showing the contrast performance of rGO/Fe3O4, rGO/Au, and rGO/Au/Fe3O4 multifunctional probes. (a) T2‐weighted image (TE = 30 milliseconds) of rGO/Fe3O4 nanosacks in CMC gel. Control (gel alone) is on the right, with nanosack concentration increasing right to left: 0.5, 1, 2, 5, 10, 20, 50 µg/mL. (b) T2 map computed from the 24 echo image series. Color bar is T2 in seconds. (c) MRI T2 map of the G/Au/Fe3O4 hybrid. Sack concentrations increase from left to right as 0.05, 0.1, 0.5, 1, 5, 10, 50, 100, and 500 µg/mL. (d) X‐ray CT image (80 kVp) of the G/Au nanosacks (right‐hand L‐shaped sequence) and free Au controls (left‐hand L‐shaped sequence). For the G/Au nanosacks, the sack concentrations in µg/mL from left to right are 2000, 400, 200, 40, 20, gel control, H2O control. For the free Au, the particle concentrations are (left to right) 400, 200, 40, 20, gel control with the 2000 µg/mL sample at the top, and a 200 µg/mL sample of empty sacks immediately beneath it. (e) Table of CT results shown as change in CT number with the attenuation (relative to control gel sample) shown in parentheses. (Reprinted with permission from Ref . Copyright 2012 American Chemical Society)
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FGO Magnetic properties. (a) SQUID magnetization curve. (b) Spin‐spin (T2) relaxation measurements obtained on a 4.7 T MRI for FGO (I: a = 625, c = 500; II: a = 313, c = 250; III: a = 156, c = 125 µg/mL) and GO (I: b = 625, d = 500; II: b = 313, d = 250; III: b = 156, d = 125 µg/mL) with the positive control (*) consisting of diluted magnevist (0.5 mg mL−1). (c) Clockwise rotational magnetic field snapshots of FGO under a field of 12 mT. (Reprinted with permission from Ref . Copyright 2013 Wiley)
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In vivo positron emission tomography/computed tomography (PET/CT) imaging of 64Cu‐labeled GO conjugates in 4T1 murine breast tumor‐bearing mice. (a) Serial coronal PET images of 4T1 tumor‐bearing mice at different time points postinjection of 64Cu‐NOTA‐GO‐TRC105, 64Cu‐NOTA‐GO, or 64Cu‐NOTA‐GO‐TRC105 after a preinjected blocking dose of TRC105. Tumors are indicated by arrowheads. (b) Representative PET/CT images of 64Cu‐NOTA‐GO‐TRC105 in 4T1 tumor‐bearing mice at 16 h postinjection. (Reprinted with permission from Ref . Copyright 2012 American Chemical Society)
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An overview of chemical structures and grafting strategies of graphene‐organic dye conjugates being used in optical imaging.
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