![]() ![]() Overall, these NE compositionsĬan be used with a library of surface attachable ligand/moieties for To the nanoemulsion surfactant coat and present the reactive cyclooctyne 25− 30 Here, we modify the dibenzocyclooctyne succinimidyl ester (DBCO-NHSĮster) linker with a lipid tail, dioctadecylamine, to facilitate incorporation Such as radioisotope labeling, live-cell imaging, and surface modificationīut has not been adapted for use with nanoemulsions. Has been used successfully for a variety of biological applications, This click chemistry occurs in mild reaction conditions and has low We demonstrate a strategy commonly known as strain-promotedĪlkyne–azide cycloaddition (SPAAC) to facilitate NE conjugation 16, 23 However, current methods for preparing surface-functionalized nanoemulsionsĪre time intensive and require a multistep process.įunctionalization, we present a novel methodologyįor efficient moiety attachment using copper-free click chemistryĪnd click-ready NE. 22 Functionalized NEsīind successfully to targets and are stable in proper storage conditions. Limitations with this strategy, such as the maleimide-linked productsīeing sensitive to hydrolysis, instability in vivo, 21 and the chemistry optimally requiringĪn inert atmosphere. Of covalent thioether bonds between maleimide and cysteine groups Sulfhydryl-based linker chemistry for NE conjugation by the formation 3 This requires chemical modification of the surface of theĭroplets to introduce functional groups, e.g., amine, carboxyl, or Surface after the emulsification process is complete. In the post-emulsification approach, the ligand is conjugated to the To withstand the extreme shear forces imposed by the homogenization. Method may be simplistic to implement, ligands must be robust enough Microfluidization) to form the stabilized NE. 16 Pre-emulsification techniques rely on ligandĬonjugation before the high-pressure, homogenization process (i.e., Two distinct methods, including pre- and post-emulsification approaches. 14, 15 Strategies for NE surface functionalization have been reported using Polymers, dyes, and chelates are of the greatest interest. Probes harboring a variety of ligand types such as peptides, antibodies, Our goal is to adorn NE with ligands bearingĬellular biomarkers as well as moieties for multimodal imaging. 11, 12 Overall, 19F MRI detection is feasible when 10 3–10 5 cells locally accumulate to generate an image hot-spot. The utility of 19F MRI has beenĭemonstrated for imaging stem cells, 2, 8 various immuneĬell subsets, 9, 10 macrophage-associated inflammatoryĭiseases, and regenerative medicine. NE is injected intravenously, and droplets are taken up by phagocyticĬells of the reticuloendothelial system (RES), including macrophagesĪt sites of inflammation. Of emerging cytotherapies used to treat cancer. This approach is particularly useful for following the fate Cell uptakeĬan be achieved by simple coincubation with cells of interest ex vivo, followed by delivery of the labeled cells to the Suspensions of PFC droplets, stabilized using a surfactant. Most often formulated as nanoemulsions (NEs), which are colloidal To their F-dense composition, which affords high MRI sensitivity as Perfluorocarbon (PFC) oils are a common fluorine tracer agent due 3, 4 Coincident 1H MRI provides the anatomical context for image interpretation. MRI due to the relatively high sensitivity, which is comparable to 1H nuclei, and its low background in biological tissues compared Tracking is a relatively new use for MRI that is making inroads towardĪn attractive tracer probe for in vivo “hot-spot” Visualizing soft-tissue anatomical morphology at high-resolution and Imaging (MRI) is a powerful technique for noninvasively Overall, these dataĭemonstrate the potential of PFC nanoemulsions incorporating DBCOĪs a versatile platform for generating functionalized probes. Of the fluorescent PFC by macrophages was demonstrated both in vitro in cultured macrophages and in situ in an acute inflammation mouse model, where fluorescence imagingĪnd 1H/ 19F magnetic resonance imaging (MRI) The feasibility of functionalization the by click reaction. As proof-of-concept,įluorescent dye-azides were conjugated to PFC nanoemulsions, demonstrating The addition of DBCO-lipid to the surfactant in PFC nanoemulsionsĭid not affect nanoemulsion size or nanoemulsion stability. Incorporating dibenzocyclooctyne (DBCO) by enabling postemulsificationįunctionalization via a click reaction with azide-containing ligands. This work, we demonstrate the utility of perfluorocarbon (PFC) nanoemulsions Targeting moieties and payloads for multimodal image generation. That is readily modifiable to accommodate binding of different molecular We describe an in vivo imaging probe platform
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