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Comparison of SPIO and USPIO for in Vitro Labeling of Human Monocytes: MR Detection and Cell Function¹

¹ From the Image Sciences Institute, University Medical Center Utrecht, Bolognalaan 50, 3584 CJ Utrecht, the Netherlands (R.D.O.E., E.L.A.B.); and Department of Molecular Cell Biology and Immunology, VU Medical Center, Amsterdam, the Netherlands (S.M.A.v.d.P., E.D., H.E.d.V.). Received January 20, 2006; revision requested March 22; revision received April 20; accepted May 17; final version accepted September 18. Supported by the Dutch MS Research Foundation (MS 01-470) and the European Commission (HPRI-CT-2001-50028). Address correspondence to R.D.O.E. (e-mail: raoul{at}invivonmr.uu.nl).

View larger version (6K): [in this window] [in a new window] [Download PPT slide]   Figure 1a: Graphs show (a) R1 and (b) R2 relaxation rates determined at 4.7-T MR imaging for increasing concentrations of USPIOs () and SPIOs (). Linear regression analyses were performed to calculate relaxivity values r1 and r2 presented as mL/(sec · µg).

View larger version (6K): [in this window] [in a new window] [Download PPT slide]   Figure 1b: Graphs show (a) R1 and (b) R2 relaxation rates determined at 4.7-T MR imaging for increasing concentrations of USPIOs () and SPIOs (). Linear regression analyses were performed to calculate relaxivity values r1 and r2 presented as mL/(sec · µg).

View larger version (26K): [in this window] [in a new window] [Download PPT slide]   Figure 2a: (a) In vitro T2 relaxation time MR image of agar-gel-suspended monocytes labeled with USPIO and SPIO at varying iron concentrations in a 96-well plate. The vehicle-treated sample contained unlabeled monocytes. (b) Graph shows R2 relaxation rates of cell samples. (c) Graph shows T2 relaxation times of increasing concentrations of SPIO-labeled monocytes. Data in b and c are presented as mean ± standard error of the mean. * = P < .05 versus vehicle-treated cells, # = P < .05 versus USPIO-labeled cells, 128e0.67x = 128 · ex.

View larger version (16K): [in this window] [in a new window] [Download PPT slide]   Figure 2b: (a) In vitro T2 relaxation time MR image of agar-gel-suspended monocytes labeled with USPIO and SPIO at varying iron concentrations in a 96-well plate. The vehicle-treated sample contained unlabeled monocytes. (b) Graph shows R2 relaxation rates of cell samples. (c) Graph shows T2 relaxation times of increasing concentrations of SPIO-labeled monocytes. Data in b and c are presented as mean ± standard error of the mean. * = P < .05 versus vehicle-treated cells, # = P < .05 versus USPIO-labeled cells, 128e0.67x = 128 · ex.

View larger version (12K): [in this window] [in a new window] [Download PPT slide]   Figure 2c: (a) In vitro T2 relaxation time MR image of agar-gel-suspended monocytes labeled with USPIO and SPIO at varying iron concentrations in a 96-well plate. The vehicle-treated sample contained unlabeled monocytes. (b) Graph shows R2 relaxation rates of cell samples. (c) Graph shows T2 relaxation times of increasing concentrations of SPIO-labeled monocytes. Data in b and c are presented as mean ± standard error of the mean. * = P < .05 versus vehicle-treated cells, # = P < .05 versus USPIO-labeled cells, 128e0.67x = 128 · ex.

View larger version (119K): [in this window] [in a new window] [Download PPT slide]   Figure 3: Detected iron oxides in monocytes with Prussian blue staining. Iron oxide is blue, and cell nuclei are red. A, Vehicle-treated monocytes. B, Monocytes incubated with USPIOs at a concentration of 1.0 mg Fe/mL. C, Monocytes incubated with SPIOs at a concentration of 1.0 mg Fe/mL. D, Higher magnification of SPIO-labeled monocytes. (Prussian blue stain; original magnification, x40 [A–C], x100 [D].)

View larger version (10K): [in this window] [in a new window] [Download PPT slide]   Figure 4: Graph shows cell viability measured by using trypan blue exclusion for monocytes labeled in the presence of USPIOs () or SPIOs () at varying iron concentrations for 1.5 hours. Data are presented as mean ± standard error of the mean. * = P < .05 versus vehicle-treated cells.

View larger version (15K): [in this window] [in a new window] [Download PPT slide]   Figure 5: Graph shows percentage of total number of added monocytes that migrated across monolayers of brain endothelium after SPIO labeling at a concentration of 1.0 mg Fe/mL. Freshly isolated monocytes (Ctrl) and vehicle-treated cells served as negative controls. Monocytes stimulated with lipopolysaccharide (LPS) for 24 hours served as a positive control. Data are presented as mean ± standard error of the mean. * = P < .05 versus freshly isolated monocytes.

View larger version (13K): [in this window] [in a new window] [Download PPT slide]   Figure 6: Graph shows IL-1 and IL-6 production of labeled monocytes 24 hours after incubation with SPIOs at a concentration of 1.0 mg Fe/mL. Freshly isolated monocytes (Ctrl) served as negative control. Monocytes stimulated with lipopolysaccharide (LPS) for 24 hours served as a positive control. Data are presented as mean ± standard error of the mean. * = P < .05 versus freshly isolated monocytes.