3 research outputs found

    J774A.1 cell studies demonstrate uptake of paramagnetic and fluorescent HDL by macrophages <i>in vitro</i>.

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    <p>J774A.1 macrophages were incubated for 2 h at 37°C with medium only or with medium containing 2.0 μM (as calculated for Rhodamine B) paramagnetic and Rhodamine B-labeled discoidal HDL (dHDL) or spherical HDL (sHDL) synthesized using a 1:1 mixture of oxidized synthetic apo A-I peptides H4 and H6. (A) Fluorescence intensities of cell lysates were measured and normalized to total cell protein content (mean ± SD, n = 3). (B) Loosely packed cell pellets were generated and <i>T</i><sub>1</sub> values were measured using <i>T</i><sub>1</sub>-weighted MR imaging. Shown in the inset are <i>T</i><sub>1</sub>-weighted images of cell pellets incubated with medium alone (M), dHDL (D) or sHDL (S). (C) Normalized enhancement ratio (NER) values for cell pellets are calculated from corresponding <i>T</i><sub>1</sub>-weighted images and are relative to cells incubated with medium only. (D) Contrast-to-noise ratio (CNR) values for cell pellets are calculated from the corresponding <i>T</i><sub>1</sub>-weighted images and are relative to cells incubated with medium only.</p

    The organ clearance depends on particle shape and composition.

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    <p>The relative signal-to-noise-ratio (SNR) increase values were calculated for regions of interest in the kidney (A) and the liver (B) of apo E KO mice at 4 h, 24 h, 48 h, 72 h following injection of the equivalent of 0.05 mmol Gd kg <sup>−1</sup> of paramagnetic and Rhodamine B-labeled discoidal HDL (dHDL) or spherical HDL (sHDL) containing a 1:1 mixture of oxidized synthetic apo A-I peptides H4 and H6. The data were calculated for each of the timepoints for a representative mouse receiving either dHDL or sHDL using the corresponding <i>T</i><sub>1</sub>-weighted images and are represented as percent SNR increase relative to the baseline pre-contrast scan (mean ± SD). Statistical difference between the 4 h and 72 h time points is shown.</p

    Confocal microscopy confirms the targeted delivery of both lipid and peptide components of GBCA-HDL to the cytoplasm of J774A.1 macrophages.

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    <p>J774A.1 cells were incubated for 2 h at 37°C with medium only or with medium containing 2.0 μM (as calculated for Rhodamine B) paramagnetic and Rhodamine B-labeled discoidal HDL (dHDL) or spherical HDL (sHDL) synthesized using a 1:1 mixture of oxidized synthetic apo A-I peptides H4 and H6. A small portion of the peptide H4 in the dHDL and sHDL preparations was fluorescently labeled with Dylight 488. (A) Differential interference contrast (DIC) image of a single representative J774A.1 macrophage incubated with sHDL (similar images were obtained for dHDL). The cell was stained with 4’,6-diamino-2-phenylindole (DAPI) dye (blue, B), visualized for Dylight 488-labeled peptide H4 (green, C) and Rhodamine B-labeled lipid (red, D). (E) The merged image shows colocalization of Dylight 488-labeled peptide H4 with Rhodamine B-labeled lipid in the cytoplasm of J774A.1 macrophages, indicating specific uptake of intact sHDL particles into the cytoplasm. White scale bar = 7.5 μM.</p
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