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Manufacture of IRDye800CW-coupled Fe3O4 nanoparticles and their applications in cell labeling and in vivo imaging.

Hou Y, Liu Y, Chen Z, Gu N, Wang J - J Nanobiotechnology (2010)

Bottom Line: The results revealed that the cells were efficiently labeled by the nanoparticles, without any significant effect on cell viability.The results demonstrated that the nanoparticles gradually accumulated in liver and kidney regions following injection, reaching maximum concentrations at 6 h post-injection, following which they were gradually removed from these regions.Real-time live-body imaging effectively reported the dynamic process of the biodistribution and clearance of the nanoparticles in vivo.

View Article: PubMed Central - HTML - PubMed

Affiliation: State key Laboratory of Bioelectronics, Southeast University, Nanjing 210096, China. wangjinke@seu.edu.cn.

ABSTRACT

Background: In recent years, near-infrared fluorescence (NIRF)-labeled iron nanoparticles have been synthesized and applied in a number of applications, including the labeling of human cells for monitoring the engraftment process, imaging tumors, sensoring the in vivo molecular environment surrounding nanoparticles and tracing their in vivo biodistribution. These studies demonstrate that NIRF-labeled iron nanoparticles provide an efficient probe for cell labeling. Furthermore, the in vivo imaging studies show excellent performance of the NIR fluorophores. However, there is a limited selection of NIRF-labeled iron nanoparticles with an optimal wavelength for imaging around 800 nm, where tissue autofluorescence is minimal. Therefore, it is necessary to develop additional alternative NIRF-labeled iron nanoparticles for application in this area.

Results: This study manufactured 12-nm DMSA-coated Fe3O4 nanoparticles labeled with a near-infrared fluorophore, IRDye800CW (excitation/emission, 774/789 nm), to investigate their applicability in cell labeling and in vivo imaging. The mouse macrophage RAW264.7 was labeled with IRDye800CW-labeled Fe3O4 nanoparticles at concentrations of 20, 30, 40, 50, 60, 80 and 100 μg/ml for 24 h. The results revealed that the cells were efficiently labeled by the nanoparticles, without any significant effect on cell viability. The nanoparticles were injected into the mouse via the tail vein, at dosages of 2 or 5 mg/kg body weight, and the mouse was discontinuously imaged for 24 h. The results demonstrated that the nanoparticles gradually accumulated in liver and kidney regions following injection, reaching maximum concentrations at 6 h post-injection, following which they were gradually removed from these regions. After tracing the nanoparticles throughout the body it was revealed that they mainly distributed in three organs, the liver, spleen and kidney. Real-time live-body imaging effectively reported the dynamic process of the biodistribution and clearance of the nanoparticles in vivo.

Conclusion: IRDye800CW-labeled Fe3O4 nanoparticles provide an effective probe for cell-labeling and in vivo imaging.

No MeSH data available.


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Prussian blue staining of cells. The agglomerates of Fe3O4 nanoparticles are stained in blue.
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Figure 2: Prussian blue staining of cells. The agglomerates of Fe3O4 nanoparticles are stained in blue.

Mentions: Cell labeling with iron nanoparticles is very important for biomedical applications [36]. Therefore, this study firstly investigated the applicability of IRDy800CW-MNPs in this field. The macrophage is commonly used as a cellular model to evaluate intravascularly administered agents, especially as it phagocytoses nanoparticles [10]. Therefore, this study employed the mouse macrophage RAW264.7 cell line to perform a cell-labeling assay. The cells were labeled with nanoparticles at various concentrations for 24 h. The cell labeling effect was evaluated by staining cells with Prussian blue and measuring the iron-loading of cells. The Prussian blue staining showed that the cells were effectively labeled by the MNPs and the IRDy800CW-MNPs (Figure 2). The blue-stained agglomerates of the iron nanoparticles in cells increased with the dose of nanoparticles in the cell culture media (Figure 2), which was in accordance with the results of the quantitative measurements of the relative iron-loading of cells using colorimetric and NIRF assays (Figure 3). In comparison, the NIRF assay reported the cellular iron-loading more sensitively than the normal colorimetric assay [69-72].


Manufacture of IRDye800CW-coupled Fe3O4 nanoparticles and their applications in cell labeling and in vivo imaging.

Hou Y, Liu Y, Chen Z, Gu N, Wang J - J Nanobiotechnology (2010)

Prussian blue staining of cells. The agglomerates of Fe3O4 nanoparticles are stained in blue.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2984479&req=5

Figure 2: Prussian blue staining of cells. The agglomerates of Fe3O4 nanoparticles are stained in blue.
Mentions: Cell labeling with iron nanoparticles is very important for biomedical applications [36]. Therefore, this study firstly investigated the applicability of IRDy800CW-MNPs in this field. The macrophage is commonly used as a cellular model to evaluate intravascularly administered agents, especially as it phagocytoses nanoparticles [10]. Therefore, this study employed the mouse macrophage RAW264.7 cell line to perform a cell-labeling assay. The cells were labeled with nanoparticles at various concentrations for 24 h. The cell labeling effect was evaluated by staining cells with Prussian blue and measuring the iron-loading of cells. The Prussian blue staining showed that the cells were effectively labeled by the MNPs and the IRDy800CW-MNPs (Figure 2). The blue-stained agglomerates of the iron nanoparticles in cells increased with the dose of nanoparticles in the cell culture media (Figure 2), which was in accordance with the results of the quantitative measurements of the relative iron-loading of cells using colorimetric and NIRF assays (Figure 3). In comparison, the NIRF assay reported the cellular iron-loading more sensitively than the normal colorimetric assay [69-72].

Bottom Line: The results revealed that the cells were efficiently labeled by the nanoparticles, without any significant effect on cell viability.The results demonstrated that the nanoparticles gradually accumulated in liver and kidney regions following injection, reaching maximum concentrations at 6 h post-injection, following which they were gradually removed from these regions.Real-time live-body imaging effectively reported the dynamic process of the biodistribution and clearance of the nanoparticles in vivo.

View Article: PubMed Central - HTML - PubMed

Affiliation: State key Laboratory of Bioelectronics, Southeast University, Nanjing 210096, China. wangjinke@seu.edu.cn.

ABSTRACT

Background: In recent years, near-infrared fluorescence (NIRF)-labeled iron nanoparticles have been synthesized and applied in a number of applications, including the labeling of human cells for monitoring the engraftment process, imaging tumors, sensoring the in vivo molecular environment surrounding nanoparticles and tracing their in vivo biodistribution. These studies demonstrate that NIRF-labeled iron nanoparticles provide an efficient probe for cell labeling. Furthermore, the in vivo imaging studies show excellent performance of the NIR fluorophores. However, there is a limited selection of NIRF-labeled iron nanoparticles with an optimal wavelength for imaging around 800 nm, where tissue autofluorescence is minimal. Therefore, it is necessary to develop additional alternative NIRF-labeled iron nanoparticles for application in this area.

Results: This study manufactured 12-nm DMSA-coated Fe3O4 nanoparticles labeled with a near-infrared fluorophore, IRDye800CW (excitation/emission, 774/789 nm), to investigate their applicability in cell labeling and in vivo imaging. The mouse macrophage RAW264.7 was labeled with IRDye800CW-labeled Fe3O4 nanoparticles at concentrations of 20, 30, 40, 50, 60, 80 and 100 μg/ml for 24 h. The results revealed that the cells were efficiently labeled by the nanoparticles, without any significant effect on cell viability. The nanoparticles were injected into the mouse via the tail vein, at dosages of 2 or 5 mg/kg body weight, and the mouse was discontinuously imaged for 24 h. The results demonstrated that the nanoparticles gradually accumulated in liver and kidney regions following injection, reaching maximum concentrations at 6 h post-injection, following which they were gradually removed from these regions. After tracing the nanoparticles throughout the body it was revealed that they mainly distributed in three organs, the liver, spleen and kidney. Real-time live-body imaging effectively reported the dynamic process of the biodistribution and clearance of the nanoparticles in vivo.

Conclusion: IRDye800CW-labeled Fe3O4 nanoparticles provide an effective probe for cell-labeling and in vivo imaging.

No MeSH data available.


Related in: MedlinePlus