研究目的
Investigating whether the uptake of very small iron oxide particles (VSOP) and ferumoxytol nanoparticles by macrophages enhances their detection by optical coherence tomography (OCT) for imaging atherosclerotic plaques.
研究成果
VSOP and ferumoxytol nanoparticles significantly increase light scattering and OCT signal when taken up by macrophages, due to clustering in phagolysosomes. This multimodal approach can improve macrophage detection in atherosclerotic plaques, with potential for enhanced OCT imaging in clinical settings.
研究不足
The OCT images of cell pellets may appear noisy due to dense packing and lack of structural features like arterial walls. The study is in vitro, and results need confirmation in vivo. The OCT system was unmodified, and future modifications could enhance detection.
1:Experimental Design and Method Selection:
The study used in vitro experiments with macrophage cell lines to measure light scattering and OCT signals after nanoparticle uptake. Methods included spectrophotometry and OCT imaging to assess backscattering and attenuation.
2:Sample Selection and Data Sources:
RAW 264.7 macrophage cells were used, incubated with VSOP (1 and 2 mM Fe) and ferumoxytol (8.9 mM Fe). Data were collected from cell pellets and nanoparticle suspensions.
3:7 macrophage cells were used, incubated with VSOP (1 and 2 mM Fe) and ferumoxytol (9 mM Fe). Data were collected from cell pellets and nanoparticle suspensions. List of Experimental Equipment and Materials:
3. List of Experimental Equipment and Materials: Equipment included a fluorescence spectrophotometer (F-7000, Hitachi Ltd), OCT imaging system (OPTIS?, St Jude Medical, Inc.), spectrophotometer (Specord? 205, Analytik Jena AG), and TEM (Zeiss EM906). Materials included VSOP, ferumoxytol, DMEM, FBS, penicillin-streptomycin, PBS, and PCR tubes.
4:6). Materials included VSOP, ferumoxytol, DMEM, FBS, penicillin-streptomycin, PBS, and PCR tubes. Experimental Procedures and Operational Workflow:
4. Experimental Procedures and Operational Workflow: Macrophages were cultured, incubated with nanoparticles, washed, and pelleted. Light scattering was measured using spectrophotometry, and OCT imaging was performed on cell pellets. Iron quantification was done using a colorimetric method.
5:Data Analysis Methods:
Data were analyzed using GraphPad Prism 5.0 for statistical analysis, including one-way ANOVA with Tukey's multiple comparison test. OCT images were analyzed using the Tissue Properties tool of the ILUMIEN? OPTIS? Offline Review Workstation.
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fluorescence spectrophotometer
F-7000
Hitachi Ltd
Measuring light scattering of control macrophages, nanoparticle-labeled macrophages, and nanoparticle suspensions in synchronous wavelength scan mode.
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spectrophotometer
Specord? 205
Analytik Jena AG
Measuring light absorption of VSOP and ferumoxytol nanoparticles.
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transmission electron microscope
EM906
Carl Zeiss Meditec AG
Imaging VSOP accumulated in macrophages of atherosclerotic plaques for TEM analysis.
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OCT imaging system
OPTIS?
St Jude Medical, Inc.
Performing OCT imaging of cell pellets and analyzing backscatter and attenuation values.
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ferumoxytol
Feraheme?
AMAG Pharmaceuticals, Inc.
Used as a nanoparticle contrast agent for incubating macrophages and measuring uptake and light scattering effects.
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VSOP
Very small iron oxide particles used as a nanoparticle contrast agent for incubating macrophages and measuring uptake and light scattering effects.
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