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Targeted gold-coated iron oxide nanoparticles for CD163 detection in atherosclerosis by MRI.

Tarin C, Carril M, Martin-Ventura JL, Markuerkiaga I, Padro D, Llamas-Granda P, Moreno JA, García I, Genicio N, Plaza-Garcia S, Blanco-Colio LM, Penades S, Egido J - Sci Rep (2015)

Bottom Line: We have prepared a targeted probe based on gold-coated iron oxide nanoparticles vectorized with an anti-CD163 antibody for the specific detection of CD163 by MRI.Subsequently, the targeted probe was injected in 16 weeks old apoE deficient mice developing atherosclerotic lesions and the pararenal abdominal aorta was imaged by MRI.The accumulation of probe in the site of interest increased over time and the signal intensity decreased significantly 48 hours after the injection.

View Article: PubMed Central - PubMed

Affiliation: Laboratorio de Patología Vascular y Renal. IIS Fundación Jiménez Díaz, Universidad Autónoma. Av. Reyes Católicos 2, 28040, Madrid, Spain.

ABSTRACT
CD163 is a membrane receptor expressed by macrophage lineage. Studies performed in atherosclerosis have shown that CD163 expression is increased at inflammatory sites, pointing at the presence of intraplaque hemorrhagic sites or asymptomatic plaques. Hence, imaging of CD163 expressing macrophages is an interesting strategy in order to detect atherosclerotic plaques. We have prepared a targeted probe based on gold-coated iron oxide nanoparticles vectorized with an anti-CD163 antibody for the specific detection of CD163 by MRI. Firstly, the specificity of the targeted probe was validated in vitro by incubation of the probe with CD163(+) or (-) macrophages. The probe was able to selectively detect CD163(+) macrophages both in human and murine cells. Subsequently, the targeted probe was injected in 16 weeks old apoE deficient mice developing atherosclerotic lesions and the pararenal abdominal aorta was imaged by MRI. The accumulation of probe in the site of interest increased over time and the signal intensity decreased significantly 48 hours after the injection. Hence, we have developed a highly sensitive targeted probe capable of detecting CD163-expressing macrophages that could provide useful information about the state of the atheromatous lesions.

No MeSH data available.


Related in: MedlinePlus

Characterization of glyconanoparticles.(A) Schematic representation of the nanoparticles. (B) UV-Vis spectra of all nanoparticles showing the appearance of the characteristic plasmon of gold. Due to their different solubilities, Fe3O4 NPs and Fe3O4@Au NPs were measured in hexane solution, whereas the rest were measured in water. (C) TEM micrograph and histogram of Fe3O4 NPs. (D) TEM micrograph and histogram of Fe3O4@Au NPs. (E) TEM micrograph and histogram of Fe3O4@Au@Man/CO2H NPs. (F) TEM micrograph and histogram of NP-CD163.
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f1: Characterization of glyconanoparticles.(A) Schematic representation of the nanoparticles. (B) UV-Vis spectra of all nanoparticles showing the appearance of the characteristic plasmon of gold. Due to their different solubilities, Fe3O4 NPs and Fe3O4@Au NPs were measured in hexane solution, whereas the rest were measured in water. (C) TEM micrograph and histogram of Fe3O4 NPs. (D) TEM micrograph and histogram of Fe3O4@Au NPs. (E) TEM micrograph and histogram of Fe3O4@Au@Man/CO2H NPs. (F) TEM micrograph and histogram of NP-CD163.

Mentions: The prepared nanoparticles consisted of a gold-coated iron oxide core covered with thiol ligands bearing either a mannose or a carboxylic acid. ProtG was covalently linked through a peptide bond to the carboxylic moieties and IgG antibodies subsequently grafted onto them (Fig. 1A). Bradford analysis of the unbound protG or antibody found in each step of the functionalization showed that the amount of protG-IgG complex on each nanoparticle ranged from 1 to 2 units. Further evidence of the grafting of the antibodies on the nanoparticles was obtained by SDS-PAGE gels of digested nanoparticles which showed the typical 2 bands of IgG antibodies, at 25 KDa and 50 KDa for the light and heavy chains, respectively (See supplemental Figure S4)23. TEM micrographs showed that the average diameter of iron oxide core was 3.2 nm, which increased up to 6.1 nm after the gold coating process (Fig. 1C–F). UV-Vis spectra were measured and as expected the characteristic plasmon resonance band of gold at around 520 nm appeared in all nanoparticles after the gold coating process (Fig. 1B). Gold and iron content was obtained by ICP-OES (Inductively coupled plasma - optical emission spectroscopy) analysis on Fe3O4@Au@Man/CO2H NPs and it was found to be 59.3 ± 0.8% and 4.03 ± 0.05% in weight, respectively. Relaxivity measurements were performed at 11.7 T and room temperature resulting in r2 and r1 values of 160 mM−1s−1 and 10 mM−1s−1, respectively. The full characterization of these nanoparticles, as superparamagnetic materials and contrast agents, has been previously described by our group2223242526.


Targeted gold-coated iron oxide nanoparticles for CD163 detection in atherosclerosis by MRI.

Tarin C, Carril M, Martin-Ventura JL, Markuerkiaga I, Padro D, Llamas-Granda P, Moreno JA, García I, Genicio N, Plaza-Garcia S, Blanco-Colio LM, Penades S, Egido J - Sci Rep (2015)

Characterization of glyconanoparticles.(A) Schematic representation of the nanoparticles. (B) UV-Vis spectra of all nanoparticles showing the appearance of the characteristic plasmon of gold. Due to their different solubilities, Fe3O4 NPs and Fe3O4@Au NPs were measured in hexane solution, whereas the rest were measured in water. (C) TEM micrograph and histogram of Fe3O4 NPs. (D) TEM micrograph and histogram of Fe3O4@Au NPs. (E) TEM micrograph and histogram of Fe3O4@Au@Man/CO2H NPs. (F) TEM micrograph and histogram of NP-CD163.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4663748&req=5

f1: Characterization of glyconanoparticles.(A) Schematic representation of the nanoparticles. (B) UV-Vis spectra of all nanoparticles showing the appearance of the characteristic plasmon of gold. Due to their different solubilities, Fe3O4 NPs and Fe3O4@Au NPs were measured in hexane solution, whereas the rest were measured in water. (C) TEM micrograph and histogram of Fe3O4 NPs. (D) TEM micrograph and histogram of Fe3O4@Au NPs. (E) TEM micrograph and histogram of Fe3O4@Au@Man/CO2H NPs. (F) TEM micrograph and histogram of NP-CD163.
Mentions: The prepared nanoparticles consisted of a gold-coated iron oxide core covered with thiol ligands bearing either a mannose or a carboxylic acid. ProtG was covalently linked through a peptide bond to the carboxylic moieties and IgG antibodies subsequently grafted onto them (Fig. 1A). Bradford analysis of the unbound protG or antibody found in each step of the functionalization showed that the amount of protG-IgG complex on each nanoparticle ranged from 1 to 2 units. Further evidence of the grafting of the antibodies on the nanoparticles was obtained by SDS-PAGE gels of digested nanoparticles which showed the typical 2 bands of IgG antibodies, at 25 KDa and 50 KDa for the light and heavy chains, respectively (See supplemental Figure S4)23. TEM micrographs showed that the average diameter of iron oxide core was 3.2 nm, which increased up to 6.1 nm after the gold coating process (Fig. 1C–F). UV-Vis spectra were measured and as expected the characteristic plasmon resonance band of gold at around 520 nm appeared in all nanoparticles after the gold coating process (Fig. 1B). Gold and iron content was obtained by ICP-OES (Inductively coupled plasma - optical emission spectroscopy) analysis on Fe3O4@Au@Man/CO2H NPs and it was found to be 59.3 ± 0.8% and 4.03 ± 0.05% in weight, respectively. Relaxivity measurements were performed at 11.7 T and room temperature resulting in r2 and r1 values of 160 mM−1s−1 and 10 mM−1s−1, respectively. The full characterization of these nanoparticles, as superparamagnetic materials and contrast agents, has been previously described by our group2223242526.

Bottom Line: We have prepared a targeted probe based on gold-coated iron oxide nanoparticles vectorized with an anti-CD163 antibody for the specific detection of CD163 by MRI.Subsequently, the targeted probe was injected in 16 weeks old apoE deficient mice developing atherosclerotic lesions and the pararenal abdominal aorta was imaged by MRI.The accumulation of probe in the site of interest increased over time and the signal intensity decreased significantly 48 hours after the injection.

View Article: PubMed Central - PubMed

Affiliation: Laboratorio de Patología Vascular y Renal. IIS Fundación Jiménez Díaz, Universidad Autónoma. Av. Reyes Católicos 2, 28040, Madrid, Spain.

ABSTRACT
CD163 is a membrane receptor expressed by macrophage lineage. Studies performed in atherosclerosis have shown that CD163 expression is increased at inflammatory sites, pointing at the presence of intraplaque hemorrhagic sites or asymptomatic plaques. Hence, imaging of CD163 expressing macrophages is an interesting strategy in order to detect atherosclerotic plaques. We have prepared a targeted probe based on gold-coated iron oxide nanoparticles vectorized with an anti-CD163 antibody for the specific detection of CD163 by MRI. Firstly, the specificity of the targeted probe was validated in vitro by incubation of the probe with CD163(+) or (-) macrophages. The probe was able to selectively detect CD163(+) macrophages both in human and murine cells. Subsequently, the targeted probe was injected in 16 weeks old apoE deficient mice developing atherosclerotic lesions and the pararenal abdominal aorta was imaged by MRI. The accumulation of probe in the site of interest increased over time and the signal intensity decreased significantly 48 hours after the injection. Hence, we have developed a highly sensitive targeted probe capable of detecting CD163-expressing macrophages that could provide useful information about the state of the atheromatous lesions.

No MeSH data available.


Related in: MedlinePlus