Nephrotoxic nephritis and obstructive nephropathy: evaluation with MR imaging enhanced with ultrasmall superparamagnetic iron oxide-preliminary findings in a rat model

PURPOSE: To evaluate the role of magnetic resonance (MR) imaging enhanced with ultrasmall superparamagnetic iron oxide (USPIO) in the evaluation and differentiation of different types of nephropathies. MATERIALS AND METHODS: Two experimental rat models of nephropathies were studied: a model of nephrotoxic nephritis induced by means of intravenous injection of sheep anti-rat glomerular basement membrane serum (n = 43) and a model of obstructive nephropathy (n = 6). Imaging sessions were performed with a spectrometer operating at 4.7 T with fast low-angle shot, or FLASH, sequences. Signal intensity was measured in each kidney compartment before and 24 hours after intravenous injection of USPIO (90 micromol of iron per kilogram of body weight). MR findings were compared with histologic data and urine protein levels. RESULTS: In the nephrotoxic nephritis model 24 hours after injection of USPIO, a significant signal intensity decrease (P: <.05) was present only in the cortex where the glomerular lesions were located. In the obstructive nephropathy model, the signal intensity decrease (P: <.05) was located in all kidney compartments in response to diffuse interstitial lesions. The decrease in signal intensity was due to iron uptake by either macrophages or mesangial cells gaining endocytic activity and was correlated, in the nephrotoxic nephritis model, to the degree of proteinuria. CONCLUSION: Twenty-four-hour delayed USPIO-enhanced MR imaging may help identify and differentiate various types of nephropathies.     

MR assessment of iodinated contrast-medium-induced nephropathy in rats using ultrasmall particles of iron oxide

The purpose of this study was to determine the diagnostic value of ultrasmall particles of iron oxide (USPIO)-enhanced MR imaging at different concentrations to evaluate experimental nephropathy. This study was conducted in 23 uninephrectomized rats using a model of iodinated contrast media-induced renal failure. Eleven rats received selective intra-arterial renal administration of diatrizoate (370 mg I/m1) and were compared to two control groups, including five animals injected with isotonic saline and seven noninjected animals. MR imaging was performed 28 hours after the procedure, including T1- and T2-weighted images before and after intravenous administration of successively 5 μmol Fe/kg and 60 μmol/kg of USPIO. Results were interpreted qualitatively and quantitatively with respect to pathologic data, and differences were studied statistically. The maximal signal intensity decrease was noted in normal kidneys in cortex (?65 ± 4%) and medulla (?84 ± 5%) on T2-weighted images after injection of 60 μmol/kg of USPIO. At this dose, diseased kidneys displayed less signal intensity decrease than normal kidneys on T2-weighted images (p = .05). Moreover, qualitative analysis showed that the highest sensitivity and specificity to diagnose kidney involvement were obtained with T2-weighted MR images (75% and 91%, respectively) when 60 μmol/kg of USPIO were used (p < .01). USPIO should be useful for in vivo evaluation of the severity of experimentally induced iodinated contrast media renal impairment in animals.     

MR imaging with ultrasmall superparamagnetic iron oxide particles in experimental soft-tissue infections in rats

PURPOSE: To investigate the feasibility of macrophage magnetic resonance (MR) imaging in rats by using an experimental soft-tissue infection model. MATERIALS AND METHODS: Thirteen rats with unilateral calf-muscle infection were imaged with a 4.7-T MR imager at an early chronic stage of infection (day 4 before contrast material injection, days 4-7 after injection). Eleven animals were imaged before and 3 and 24 hours after intravenous application of ultrasmall superparamagnetic iron oxide (USPIO), and eight animals were additionally imaged 48 hours and three animals 72 hours after USPIO application. Two infected rats served as controls. T1- and T2-weighted spin-echo and T2*-weighted gradient-echo sequences were applied. All animals were sacrificed, and histopathologic findings were correlated with findings on MR images. Electron microscopy was performed in two rats. For quantitative analysis, signal intensities on T2*-weighted images and T2 values on T2 maps were measured within regions of interest, and the temporal variation was analyzed by using the signed rank test. RESULTS: Visualization of USPIO-loaded macrophages was most sensitive with a T2*-weighted sequence. USPIO distribution pattern and quantitative analysis of T2 and T2* effects 3 hours after USPIO application were significantly different (P <.05) from those at 24 and 48 hours, reflecting the dynamic transit of the particle accumulation from the intravascular to the intracellular compartment by means of macrophage phagocytosis. Local signal intensity alterations could be correlated with iron-loaded macrophages at histopathologic examination. CONCLUSION: Activated macrophages in acute soft-tissue infection can be labeled with USPIOs and detected with MR imaging because of susceptibility effects.     

USPIO-enhanced MR imaging of macrophage infiltration in native and transplanted kidneys: initial results in humans

The purpose of this study was to evaluate the detection and characterization of macrophage infiltration in native and transplanted kidneys using ultrasmall superparamagnetic iron oxide particles (USPIO). Among 21 patients initially enrolled, 12 scheduled for renal biopsy for acute or rapidly progressive renal failure (n = 7) or renal graft rejection (n = 5) completed the study. Three magnetic resonance (MR) sessions were performed with a 1.5-T system, before, immediately after and 72 h after i.v. injection of USPIO at doses of 1.7–2.6 mg of iron/kg. Signal intensity change was evaluated visually and calculated based on a region of interest (ROI) positioned on the kidney compartments. Histological examination showed cortical macrophage infiltration in four patients (>5 macrophages/mm²), two in native kidneys (proliferative extracapillary glomerulonephritis) and two in transplants (acute rejection). These patients showed a 33 ± 18% mean cortical signal loss on T2*-weighted images. In the remaining eight patients, with <5 macrophages/mm², there was no cortical signal loss. However, in three of these, presenting with ischemic acute tubular necrosis, a strong (42 ± 18%) signal drop was found in the medulla exclusively. USPIO-enhanced MR imaging can demonstrate infiltration of the kidneys by macrophages both in native and transplanted kidneys and may help to differentiate between kidney diseases.     

MR receptor imaging: ultrasmall iron oxide particles targeted to asialoglycoprotein receptors

Previously we have reported that ultrasmall superparamagnetic iron oxide (USPIO) particles migrate across capillary endothelium, a prerequisite for the design of particulate pharmaceuticals for MR receptor imaging. In the current study, USPIO particles are directed specifically to asialoglycoprotein (ASG) receptors by coupling galactose terminals in the form of arabinogalactan (AG) to these particles. Biodistribution data showed that ASG-directed, AG-coated USPIO (AG-USPIO) particles selectively accumulate in the liver but not in other organs. Electron microscopy of liver showed electron-dense iron oxide particles bound to hepatocyte cell-surface membranes and in large numbers within intracellular lysosomes. The specificity of AG-USPIO for asialoglycoprotein receptors was confirmed by incubation experiments with and without ASG-blocking agents such as D(+)galactose and asialofetuin. In vivo MR imaging in rats showed a significant decrease in liver signal intensity at low doses (2 mumol Fe/kg); no significant changes were observed in the spleen. This decrease in signal intensity is larger than that observed with conventional iron oxides at equal doses. These initial data suggest that, for the first time, superparamagnetic agents can be directed to specific sites for MR imaging by strategies such as receptor targeting.     

MR imaging of relapsing multiple sclerosis patients using ultra-small-particle iron oxide and compared with gadolinium

BACKGROUND AND PURPOSE: Inflammatory multiple sclerosis (MS) lesions are characterized by microglia activation and infiltration of T cells, B cells, and macrophages across the blood-brain barrier (BBB). In the experimental autoimmune encephalomyelitis (EAE) rat model of MS, previous MR imaging investigations with a new contrast agent ultra-small-particle iron oxide (USPIO) that accumulates in phagocytic cells revealed in vivo the presence of macrophage brain infiltration. The goal of this study was to characterize MS lesions with the use of this contrast agent. METHODS: A prospective MR imaging study of 10 patients with MS in acute relapses was achieved by using USPIO and gadolinium. RESULTS: Twenty-four hours after USPIO injection, 33 acute MS lesions in 9 patients showed USPIO uptake. Lesions were seen as high signal intensities on T1-weighted images and low signal intensities on T2-weighted images. Gadolinium enhancement was seen in 31 of these lesions in 7 patients. These 7 patients presented 24 gadolinium-enhanced lesions that did not enhance with USPIO. Two patients showed USPIO-enhanced lesions but no gadolinium-enhanced lesions. CONCLUSION: Taken together with earlier findings obtained in experimental models or in human stroke, the visualization of macrophage activity in vivo with USPIO characterize a distinct cellular and inflammatory event of the dynamic process of MS lesion formation. The macrophage activity information obtained with USPIO is distinct and complementary to the increased BBB permeability seen with gadolinium.     

Targeting of ultrasmall superparamagnetic iron oxide (USPIO) particles to tumor cells in Vivo by using transferrin receptor pathways

Human transferrin was covalently coupled to ultrasmall superparamagnetic iron oxide (USPIO) particles, and the trans-ferrin-USPIO obtained was investigated in vivo in experimental SMT/2A tumor-bearing rats (rat mammary carcinoma). Physicochemical characterization showed an overall size of 36 nm (DLS) with a core size of 5 nm (TEM). Relaxivities were R,₁ = 23.6 and R₂ = 52.1 liter/mmol · s (0.47 T). Bound transferrin was 280 μg/mg of iron. Pharmacokinetic investigations revealed a half-life of 17 min in normal rats. The MR evaluation of tumor signal intensity over time showed a 40% (range 25–55%) signal reduction 150 min after injection with the reduction persisting for at least 8 h. Control experiments using the parent USPIO compound or USPIO labeled with a nonspecific human serum albumin (HSA-USPIO) showed a change of only 10% (range 5–15%) in tumor signal intensity over time. The results demonstrate that a combination of the USPIO relaxivity properties with the specificity of transferrin-medi-ated endocytosis allows in vivo detection of tumors by MR imaging.     

Use of a blood-pool contrast agent for MR-guided vascular procedures: feasibility of ultrasmall superparamagnetic iron oxide particles

RATIONALE AND OBJECTIVES: The purpose of this study was to examine the dose dependency of the intravascular signal intensity after injection of ultrasmall superparamagnetic iron oxide (USPIO) particles (SH U 555 C) in a rabbit model studied with a low-field-strength magnetic resonance (MR) imaging system. The results were used to facilitate MR-guided vascular procedures in a pig. MATERIALS AND METHODS: All experiments were performed at 0.2 T. To determine the optimum USPIO (or SH U 555 C) dose for intravascular interventions, the authors acquired coronal three-dimensional MR angiographic images in 12 rabbits after injection of four dose levels (10, 20, 30, and 40 micromol of iron per kilogram body weight). The intraaortic signal intensities were measured in user-defined regions of interest. For numerical analysis, signal intensity enhancement was computed. Subsequently MR image-guided procedures were performed in USPIO-enhanced vessels in one pig. RESULTS: The signal intensity evaluation shows a clear-cut dose dependence in both early and late phases after administration of SH U 555 C. A high-spatial-resolution MR angiogram acquired 20 minutes after injection yielded the best results with the highest dose (40 micromol of iron per kilogram); at that dose, intravascular enhancement was sufficient for vascular procedures for 60 minutes after injection. CONCLUSION: SH U 555 C is a promising contrast agent for MR angiography and MR-guided vascular procedures in an open low-field-strength MR imager.     

In vitro characterization of two different ultrasmall iron oxide particles for magnetic resonance cell tracking

RATIONALE AND OBJECTIVES: Comparison of two different ultrasmall superparamagnetic iron oxide (USPIO) particles in terms of their intracellular cell-labeling properties of macrophages and subsequent visualization by MR imaging. MATERIALS AND METHODS: Cultures containing the macrophage cell line P-388D1 were incubated with a neutral carboxydextran-coated USPIO preparation (DDM 43/34/103) or an acidic citrate-coated USPIO (VSOP-C125). Experiments were performed in which incubation concentration and duration were varied and phagocytosis and pinocytosis suppressed by specific inhibitors. In cell culture specimens iron content was measured quantitatively and signal intensities determined by in vitro MR imaging. RESULTS: VSOP-C125 is incorporated by cells much faster than DDM 43/34/103 and produces significantly higher final intracellular iron concentrations per cell (3420 vs. 727 ng/million cells). Both preparations show similar signal-reducing effects at MR imaging relative to the Fe content per cell. Intracellular USPIO has a much lower detection threshold at MR imaging (50/80 micromol/L) than extracellular USPIO in free solution (300 micromol/L). CONCLUSIONS: Citrate-coated USPIO particles VSOP-C125 appear to have more favorable properties for magnetic labeling of macrophages than the carboxydextran-coated USPIO preparation DDM 43/34/103.     

Ultrasmall superparamagnetic iron oxide: an intravenous contrast agent for assessing lymph nodes with MR imaging

An ultrasmall superparamagnetic iron oxide (USPIO) preparation was evaluated as a potential intravenous contrast agent for lymph nodes. Relaxation time measurements and magnetic resonance (MR) imaging were performed in rats with normal lymph nodes and in rats with lymph node metastases. In normal animals, lymph node relaxation times decreased maximally within 24-48 hours after intravenous administration of USPIO. Twenty-four hours after administration, the T2 of normal lymph nodes had decreased from 74 msec +/- 2.2 to 30 msec +/- 0.7 (USPIO, 40 mumol of iron per kilogram) or 15 msec +/- 0.0 (200 mumol Fe/kg), whereas the T2 of metastatic nodes did not change. MR imaging of the animal model of nodal metastases confirmed the hypothesis that intravenously administered USPIO decreases signal intensity of normal but not metastatic nodes. A single intravenous administration of USPIO may allow detection of nodal metastases on the basis of signal intensity characteristics rather than the currently used, insensitive size characteristics.     

Iron oxide-enhanced MR lymphography: initial experience

The detection of nodal metastases is of utmost importance in oncologic imaging. Ultrasmall superparamagnetic iron oxide particles (USPIO) are novel contrast agents specifically developed for MR lymphography. After intravenous administration, they are taken up by the macrophages of the lymph nodes, where they accumulate. They reduce the signal intensity (SI) of normally functioning nodes on postcontrast T2-and T2*-weighted images through the magnetic susceptibility effects on iron oxide. Metastatic nodes, in which macrophages are replaced by tumor cells, show no significant change in SI on postcontrast T2-and T2*-weighted images. Early clinical experience suggests that USPIO-enhanced MR lymphography improves the sensitivity and specificity for the detection of nodal metastases. It also suggests that micrometastases could be detected in normal-sized nodes. This article reviews the physiochemical properties of USPIO contrast agents, their enhancement patterns, and early clinical experience.     

Comparison of ultrasmall particles of iron oxide (USPIO)-enhanced T2-weighted, conventional T2-weighted, and gadolinium-enhanced T1-weighted MR images in rats with experimental autoimmune encephalomyelitis

BACKGROUND AND PURPOSE: Ultrasmall particles of iron oxide (USPIO) constitute a contrast agent that accumulates in cells from the mononuclear phagocytic system. In the CNS they may accumulate in phagocytic cells such as macrophages. The goal of this study was to compare USPIO-enhanced MR images with conventional T2-weighted images and gadolinium-enhanced T1-weighted images in a model of experimental autoimmune encephalomyelitis (EAE). METHODS: Nine rats with EAE and four control rats were imaged at 4.7 T and 1.5 T with conventional T1- and T2-weighted sequences, gadolinium-enhanced T1-weighted sequences, and T2-weighted sequences obtained 24 hours after intravenous injection of a USPIO contrast agent, AMI-227. Histologic examination was performed with hematoxylin-eosin stain, Perls' stain for iron, and ED1 immunohistochemistry for macrophages. RESULTS: USPIO-enhanced images showed a high sensitivity (8/9) for detecting EAE lesions, whereas poor sensitivity was obtained with T2-weighted images (1/9) and gadolinium-enhanced T1-weighted images (0/9). All the MR findings in the control rats were negative. Histologic examination revealed the presence of macrophages at the site where abnormalities were seen on USPIO-enhanced images. CONCLUSION: The high sensitivity of USPIO for macrophage activity relative to other imaging techniques is explained by the histologic findings of numerous perivascular cell infiltrates, including macrophages, in EAE. This work supports the possibility of intracellular USPIO transport to the CNS by monocytes/macrophages, which may have future implications for imaging of human inflammatory diseases.     

USPIO-enhanced MRI of highly invasive and highly metastasizing transplanted human squamous cell carcinoma: an experimental study

Objective

The aim of this study was to investigate the signal intensity characteristics of highly invasive and highly metastasizing transplanted human squamous cell carcinoma using ultra-small super-paramagnetic iron oxide (USPIO)-enhanced MRI and to correlate them with USPIO distribution to tumour components revealed by histological examination.

Methods

13 nude mice with transplanted human squamous cell carcinoma in the oral cavity were imaged before and 24 hours after intravenous administration of USPIO. The difference in signal intensity between pre-contrast and post-contrast MR images was visually evaluated. For quantitative analysis, signal intensity within a region of interest was measured. Histological findings were correlated with MR findings. The approximate USPIO concentration was evaluated using USPIO phantoms.

Results

Seven tumours had an area showing signal intensity increase on post-contrast T₁ weighted images. Histopathologically, six of those tumours contained a small amount of iron particles in the stroma. The USPIO concentration was presumed low. Two tumours had an area showing signal intensity decrease on post-contrast T₁ and T₂ weighted images. The areas had a large amount of iron particles in the stroma and the USPIO concentration was presumed high. There was a minimal amount of iron particles in tumour parenchymal cells.

Conclusions

The amount of USPIO accumulation into tumour stroma was considered to affect MR signal intensity. A small amount increases T₁ weighted signal intensity, whereas a large amount decreases T₁ and T₂ weighted intensity. The USPIO accumulation into the tumour parenchyma was not thought to affect MR signal intensity.     

Bone marrow: ultrasmall superparamagnetic iron oxide for MR imaging

An ultrasmall superparamagnetic iron oxide (USPIO) preparation was evaluated as a potential intravenous contrast agent for magnetic resonance (MR) imaging of bone marrow. One hour after administration of USPIO (40, 80, and 160 mumols of iron per kilogram body weight) in rats and rabbits, T1 and T2 relaxation times were, respectively, approximately 30%, 50%, and 65% lower than precontrast relaxation times. Maximum decrease in relaxation times of marrow occurred within 1-24 hours after intravenous administration; thereafter, relaxation times slowly returned to normal within 7 days. In vivo MR imaging of rabbits and rats confirmed that USPIO decreases signal intensity of red and yellow marrow. The decrease was most marked with gradient echo pulse sequences. An animal model of intramedullary tumor demonstrated the potential of USPIO to enable differentiation between tumor and normal red marrow. USPIO-enhanced MR imaging improves detection of smaller tumors and allows differentiation of tumor deposits from islands of hyperplastic or normal red marrow.     

Detection of synovial macrophages in an experimental rabbit model of antigen-induced arthritis: ultrasmall superparamagnetic iron oxide-enhanced MR imaging

PURPOSE: To evaluate intravenously administered ultrasmall superparamagnetic iron oxide (USPIO) as a marker of macrophage activity in an experimental rabbit model of antigen-induced arthritis. MATERIALS AND METHODS: Unilateral arthritis was induced by means of intraarticular injection of methylated bovine serum albumin in 10 knees of 10 rabbits that had been presensitized to the same antigen. The contralateral knees in these rabbits, as well as six knees in three other rabbits, served as controls. After onset of arthritis, all knees were imaged prior to and 24 hours after administration of USPIO. The magnetic resonance (MR) imaging protocol included T1-weighted spin-echo, T2-weighted fast spin-echo, T2*-weighted gradient-echo, and short inversion time inversion-recovery sequences. Images were analyzed quantitatively and qualitatively with regard to signal characteristics and pattern. MR findings were correlated with histopathologic findings. Wilcoxon signed rank test was used to compare results of signal-to-noise ratio calculations before and after USPIO administration. RESULTS: All knees with intraarticular injection of antigen suspension developed unilateral arthritis, whereas no signs of arthritis occurred in the control knees. On USPIO-enhanced images obtained 24 hours after contrast agent administration, significant T1 (P =.03) and more predominantly T2* (P =.02) and T2 effects (P =.01) were evident in the synovium of all 10 arthritic knees, which reflected USPIO uptake by macrophages in the synovial tissue. To a lesser extent, T2* effects were present also within the joint effusion (P =.01). No significant changes in signal characteristics were detected in the 10 nonarthritic knees in the antigen-injected group or the six knees in the control group (P =.06-.91). Histologic examination confirmed uptake of iron in the macrophages of arthritic knees. Changes in MR signal characteristics within the arthritic synovium and synovial effusion were visually detectable after intravenous administration of USPIO. CONCLUSION: MR imaging at 1.5 T can depict USPIO uptake in phagocytic-active macrophages in an antigen-induced arthritis animal model.     

Dosage determination of ultrasmall particles of iron oxide for the delineation of microvasculature in the Wistar rat brain

PURPOSE: This investigation sought to optimize ultrasmall particles of iron oxide (USPIO) contrast agent dosage for visualizing cerebral microvasculature on an 8.0-Tesla ultra high field magnetic resonance imaging system. MATERIALS AND METHODS: USPIO contrast agent was intravenously administered to 3 groups of 4 rats at 1, 2, and 3 mg Fe/kg. Each animal was scanned before and after injection of USPIO using a high resolution T2*-weighted gradient recalled echo sequence with an in-plane resolution of 78 microm. The signal-to-noise ratio (SNR) and the number of microvessels visualized within the cortex and basal ganglia were calculated and compared before and after the administration of USPIO. RESULTS: As the USPIO dose increased, microvascular conspicuity increased, and SNR decreased. A dosage of 2 mg Fe/kg improved microvascular visualization in both cortex and basal ganglion regions relative to 1 mg Fe/kg without significantly sacrificing SNR as was the case at 3 mg Fe/kg. CONCLUSION: Two mg Fe/kg USPIO is an optimal dose when imaging normal rat cerebral microvasculature using GRE T2*-weighted MR imaging at a field strength of 8 T.     

急性实验性变态反应性脑脊髓炎模型的MRI表现及其病理基础

目的 探讨急性实验性变态反应性脑脊髓炎(EAE)模型的MRI表现及其病理基础.方法 用豚鼠全脑脊髓匀浆诱导EAE大鼠模型(6只),观察其体重变化和临床表现,同时设对照组(6只).在EAE大鼠发病后进行MR常规扫描、钆剂增强扫描,并静脉使用超小超顺磁性氧化铁(USPIO),24 h后行USPIO增强扫描,扫描完毕立即处死大鼠取脑,行脑组织切片的HE染色、髓鞘染色和普鲁士蓝染色.观察MRI和病理检查的异常所见,并进行对照.结果EAE大鼠组在诱导后第8～9天开始体重下降,在第10～11天出现临床症状.MR常规扫描未见明确异常,钆剂增强后仅见脑膜弥漫增厚及增强,USPIO增强扫描可见T2WI上延髓实质内大片低信号区,T1WI可见对应部位的高信号.梯度回波T2*WI比T2WI显示在脑干病灶以外的小脑白质内低信号.临床评分高的大鼠其低信号范围也较大.对照组大鼠未见异常.病理检查发现发病大鼠脑白质内散在血管套袖,部分伴相邻区域的脱髓鞘改变.普鲁士蓝染色发现病灶内巨噬细胞胞质内有蓝染颗粒,沉积部位与T2WI上低信号对应.结论USPIO增强MRI可以揭示常规MRI和钆剂增强成像未能显示的急性EAE病变,活体显示EAE病灶内巨噬细胞的分布,具有重要的研究价值和应用前景.     

Lymph nodes: microstructural anatomy at MR imaging

High-resolution microscopic magnetic resonance (MR) images of rodent lymph nodes were directly correlated with sections obtained for histologic study to determine the microstructural anatomy of lymph nodes seen at MR imaging and to evaluate signal intensity changes induced by a novel intravenous lymphotropic MR contrast agent (ultrasmall superparamagnetic iron oxide [USPIO]). High-resolution T2-weighted images of unenhanced lymph nodes demonstrated medullary sinus as regions of low signal intensity and follicles as high-intensity structures. After a single intravenous administration of USPIO (160 mumol/kg), both T1-weighted and T2-weighted images showed areas of focal signal intensity loss in medullary sinuses corresponding to the distribution of uptake by macrophages. Lymph follicles appeared unchanged in signal intensity, as they are largely devoid of macrophages. This model of microscopic MR imaging should provide the basis for (a) understanding differences between patterns of contrast-enhanced normal lymph nodes and those of diseased ones and (b) guiding the development of targeting strategies for novel pharmaceuticals at the cellular level.     

MRI detection of macrophages labeled using micrometer-sized iron oxide particles

PURPOSE: To evaluate cellular labeling of immune cells using micron-sized iron oxide particles (MPIOs) and evaluate the MR relaxivity and MRI detection of the labeled cells. MATERIALS AND METHODS: Immune cells isolated from mice and rats were labeled with three different sizes of MPIO particles (0.35, 0.90, or 1.63 microm). These labeled cells were characterized using transmission electron microscopy (TEM), fluorescence microscopy, flow cytometry, MR relaxometry, and MRI. RESULTS: Macrophage uptake of MPIOs was found to be highest for the 1.63-microm size particles. MR relaxivity measurements indicated greater spin-spin relaxation for MPIO-labeled cells relative to cells labeled with nanometer-sized ultra-small superparamagnetic iron oxide (USPIO) particles with similar iron content. TEM and fluorescence microscopy indicated cellular uptake of multiple MPIO particles per cell. Macrophages labeled with 1.63-microm MPIOs had an average cellular iron uptake of 39.1 pg/cell, corresponding to approximately 35 particles per cell. CONCLUSION: Cells labeled with one or more MPIO particles could be readily detected ex vivo at 11.7 Tesla and after infusion of the MPIO-labeled macrophages into the kidney of a rat, hypointense regions of the outer cortex are observed, in vivo, by MRI at 4.7 Tesla.     

Macrophage infiltration detected at MR imaging in rat kidney allografts: early marker of chronic rejection?

PURPOSE: To evaluate detection of iron-loaded macrophages at magnetic resonance (MR) imaging as a noninvasive means to monitor early signs of chronic allograft rejection in the life-supporting Fisher-to-Lewis rat kidney transplantation model. MATERIALS AND METHODS: Experiments followed the Swiss federal regulations of animal protection. Male Fisher (n = 37) and Lewis (n = 77) rats were used. After removal of a native recipient kidney and transplantation of a donor kidney, the recipient rat's contralateral kidney was removed. Allografts and control syngeneic grafts comprised, respectively, kidneys from Fisher and Lewis donors transplanted into Lewis rats. Recipients were imaged by using a gradient-echo MR sequence 24 hours after intravenous administration of superparamagnetic iron oxide (SPIO) particles. Biochemical analyses of blood and urine, as well as assessments of Banff scores (reference standard for histologic classification of graft rejection), were performed. Statistical tests used were analysis of variance for multiple comparisons with Bonferroni tests, Mann-Whitney tests, and Pearson correlations with Bonferroni corrections. RESULTS: A SPIO dose-dependent decrease in cortical MR signal intensity occurred in allografts between 8 and 16 weeks after transplantation. A strong significant negative correlation (P = .005 for 0.3 mL/kg SPIO dose, P = .003 for 1.0 mL/kg SPIO dose) was found between MR signal intensity and Banff scores, which deteriorated over the experimental period. Proteinuria occurred at 16 weeks. Blood and urine creatinine levels remained unchanged up to week 28. CONCLUSION: This MR imaging method is more robust than the usually adopted creatinine clearance method for the detection of early signs of allograft chronic rejection in the Fisher-to-Lewis rat kidney transplantation model.