Contrast enhancement in experimental radiation-induced liver injury: Comparison of hepatocellular and reticuloendothelial particulate contrast agents

We compared the liver enhancement of two super-paramagnetic agents, polycrystalline iron oxide nano-particles (PION) and PION coated with asialofetuin (ASF), in an experimental model of focal radiation-induced hepatitis. PION, a reticuloendothelial system-directed agent, and PION-ASF, a hepatocellular-directed agent, were compared for time-dependent liver enhancement in an experimental model of radiation-induced liver injury. Using the reticuloendothelial system (RES)-directed PION, the normal, nonirradiated portion of the liver decreased in signal intensity (SI) with a mean negative enhancement of –66% ± 4, whereas the irradiated portion (60 Gy, 3 days before imaging) of the liver decreased in SI by –24% ± 2, significantly less (P < .05). SI changes in irradiated liver tissue using PION were dose-dependent, being more pronounced with lower radiation exposure. The difference in SI changes induced by PION-ASF between irradiated and nonirradiated liver was not statistically different, but SI decreased with a mean negative enhancement of –80% ± 2. The RES-directed PION is more sensitive for the detection of radiation-induced hepatitis than is the hepatocyte-directed PION-ASF. The insensitivity of PION-ASF enhancement for diffuse liver injury may be clinically advantageous for detecting focal lesions in the presence of diffuse hepatic injury.     

Superparamagnetic iron oxide contrast agents: physicochemical characteristics and applications in MR imaging

Superparamagnetic iron oxide MR imaging contrast agents have been the subjects of extensive research over the past decade. The iron oxide particle size of these contrast agents varies widely, and influences their physicochemical and pharmacokinetic properties, and thus clinical application. Superparamagnetic agents enhance both T1 and T2/T2* relaxation. In most situations it is their significant capacity to reduce the T2/T2* relaxation time to be utilized. The T1 relaxivity can be improved (and the T2/T2* effect can be reduced) using small particles and T1-weighted imaging sequences. Large iron oxide particles are used for bowel contrast [AMI-121 (i.e. Lumirem and Gastromark) and OMP (i.e. Abdoscan), mean diameter no less than 300 nm] and liver/spleen imaging [AMI-25 (i.e. Endorem and Feridex IV, diameter 80-150 nm); SHU 555A (i.e. Resovist, mean diameter 60 nm)]. Smaller iron oxide particles are selected for lymph node imaging [AMI-227 (i.e. Sinerem and Combidex, diameter 20-40 nm)], bone marrow imaging (AMI-227), perfusion imaging [NC100150 (i.e. Clariscan, mean diameter 20 nm)] and MR angiography (NC100150). Even smaller monocrystalline iron oxide nanoparticles are under research for receptor-directed MR imaging and magnetically labeled cell probe MR imaging. Iron oxide particles for bowel contrast are coated with insoluble material, and all iron oxide particles for intravenous injection are biodegradable. Superparamagnetic agents open up an important field for research in MR imaging.     

Assessment of T1 and T2* effects in vivo and ex vivo using iron oxide nanoparticles in steady state--dependence on blood volume and water exchange.

Accurate knowledge of the relationship between contrast agent concentration and tissue relaxation is a critical requirement for quantitative assessment of tissue perfusion using contrast-enhanced MRI. In the present study, using a pig model, the relationship between steady-state blood concentration levels of an iron oxide nanoparticle with a hydrated diameter of 12 nm (NC100150 Injection) and changes in the transverse and longitudinal relaxation rates (1/T2* and 1/T1, respectively) in blood, muscle, and renal cortex was investigated at 1.5 T. Ex vivo measurements of 1/T2* and 1/T1 were additionally performed in whole pig blood spiked with different concentrations of the iron oxide nanoparticle. In renal cortex and muscle, 1/T2* increased linearly with contrast agent concentration with slopes of 101 +/-22 s(-1)mM(-1) and 6.5 +/-0.9 s(-1)mM(-1) (mean +/- SD), respectively. In blood, 1/T2* increased as a quadratic function of contrast agent concentration, with different quadratic terms in the ex vivo vs. the in vivo experiments. In vivo, 1/T1 in blood increased linearly with contrast agent concentration, with a slope (T1-relaxivity) of 13.9 +/- 0.9 s(-1)mM(-1). The achievable increase in 1/T1 in renal cortex and muscle was limited by the rate of water exchange between the intra- and extravascular compartments and the 1/T1-curves were well described by a two-compartment water exchange limited relaxation model.     

Field dependent transverse relaxation rate increase may be a specific measure of tissue iron stores

The degree to which MRI magnet field strength affects measured transverse relaxation rates (R₂) defines a measure termed the field dependent R₂ increase (FDRI). We report here the results of in vivo and in vitro experiments that were conducted to evaluate whether FDRI is a potentially useful measure of tissue iron stores. T₂ relaxation times were obtained using two clinical MRI instruments operating at 0.5 and 1.5 Tesla, and relaxation rates (R₂) were calculated as the reciprocal of T₂. The in vivo experiment measured R₂ in human brain frontal white matter, caudate nucleus, putamen, and globus pallidus. The FDRI was very highly correlated with published brain iron levels for the four regions examined. The in vitro experiment measured R₂ in agarose gel-based phantoms containing physiologic forms and amounts proteins involved in iron storage and transport (ferritin, apoferritin, transferrin, and apotransferrin). Significant field dependence was observed only for the ferritin phantoms. The differences in the R₂ values obtained at the two field strengths were striking, and were proportional to the ferritin levels of the phantoms. These studies suggest that FDRI may be a specific measure of tissue ferritin. The quantitative significance of the results to imaging and possible applications to the clinical investigation of pathologic states are discussed.     

Overcoming the low relaxivity of gadofosveset at high field with spin locking

The contrast agent gadofosveset, which binds reversibly to serum albumin, has a high longitudinal relaxivity at lower magnetic fields (≤3.0 T) but a much lower relaxivity at high fields. Spin locking is sensitive to macromolecular content; it is hypothesized that combining this technique with the albumin‐binding properties of gadofosveset may enable increased relaxivity at high fields. In vitro measurements at 4.7 T found significantly higher spin‐lock relaxation rates, R_1ρ (1/T_1ρ), when gadofosveset was serum albumin‐bound than when unbound. R_1ρ values for a nonbinding contrast agent (gadopentetate dimeglumine) in serum albumin were similar to those for unbound gadofosveset. R₂ (1/T₂) values were also significantly higher at 4.7 T for serum albumin‐bound gadofosveset than for unbound. Spin locking at high field generates significantly higher relaxation rates for gadofosveset than conventional contrast agents and may provide a method for differentiating free and bound molecules at these field strengths. Magn Reson Med, 2012. © 2011 Wiley Periodicals, Inc.     

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.     

Superparamegnetic iron oxides for MRI

Pharmaceutical iron oxide preparations have been used as MRI contrast agents for a variety of purposes. These agents predominantly decrease T2 relaxation times and therefore cause a decrease in signal intensity of tissues that contain the agent. After intravenous adminstration, dextran-coated iron oxides typically accumulate in phagocytic cells in liver and spleen. Clinical trials have shown that iron oxide increases lesion/liver and lesion/spleen contrast, that more lesions can be depicted than on plain MRI or CT, and that the size threshold for lesion detection decreases. Decreased uptake of iron oxides in liver has been observed in hepatitis and cirrhosis, potentially allowing the assessment of organ function. More recently a variety of novel, target-specific monocrydtalline iron oxides compounds have been used for receptor and immunospecific images. Future development of targeted MRI contrast agents is critical for organ- or tissue-specific quantitative and functional MRI. Correspondence to: R. Weissleder     

Boron nitride nanotubes for boron neutron capture therapy as contrast agents in magnetic resonance imaging at 3 T

The applicability of boron nitride nanotubes (BNNTs) containing Fe paramagnetic impurities as contrast agents in magnetic resonance imaging (MRI) was investigated. The measurement of longitudinal and transverse relaxation times of water protons in homogeneous aqueous dispersions of BNNTs wrapped with poly(l-lysine) at different concentrations allowed longitudinal (r₁) and transverse (r₂) relaxivities to be determined at 3 T. The r₂ value was comparable to those of commercial superparamagnetic iron oxide nanoparticles, indicating that Fe-containing BNNTs have the potential to be used as T₂ contrast-enhancement agents in MRI at 3 T.     

Receptor imaging: application to MR imaging of liver cancer

A new contrast agent for magnetic resonance (MR) imaging, directed to asialoglycoprotein (ASG) receptors on hepatocytes, was used for detection of liver cancer in rats. Ultrasmall superparamagnetic (mean size, 12 nm) particles of iron oxide (USPIOs) were targeted to ASG receptors by coating particles with arabinogalactan (AG). Liver T2 relaxation times decreased more effectively after a single intravenous administration of AG-USPIO than after an equal dose of a conventional superparamagnetic liver MR contrast agent (AMI-25; mean size, 72 nm). Receptor affinity studies demonstrated that receptor-mediated attachment and subsequent cellular endocytosis do not occur in primary malignant (hepatocellular carcinoma) or metastatic (adenocarcinoma) tumors, because the surface ASG receptors are lost during malignant dedifferentiation. In vitro relaxation and in vivo MR imaging experiments of liver tumors show that targeting USPIO to hepatocytes rather than to the mononuclear phagocytic system allows a considerable dose reduction, increases tumor-liver contrast, and potentially allows distinction of ASG-positive (benign hepatocellular) and ASG-negative (malignant hepatocellular) tumors.     

NMR imaging of changes in vascular morphology due to tumor angiogenesis

Tumor-sprouted vessels are greater in both number and diameter in comparison to their healthy counterparts. A novel technique based on magnetic susceptibility contrast mechanisms that are sensitive to varying sizes of blood vessels is presented to measure differences between the relaxation rates (1/T₂ and 1/T) in a rat glioma model and normal cerebral cortex. ΔR2 and ΔR2*, the differences between relaxation rates precontrast and postcontrast agent injection, were measured for an intravascular equilibrium contrast agent (MION) at various echo times. Since ΔR2*/ΔR2 increases as vessel size increases, this ratio can be used as a measure of the average vessel size within an ROI or a voxel. The stability and longevity of the contrast agent within the vasculature were verified (n = 2 trials), and the ratio of ΔR2*/ΔR2 between the tumor and normal cortex was measured to be 1.9 ± 0.2 (n = 4, echo time = 20 ms, and susceptibility difference (Δχ) ≈? 10^?6). This ratio compared favorably to a predicted ratio determined using histologically determined vessel sizes and theoretical Monte Carlo modeling results (1.9 ± 0.1). Maps of the ratio of ΔR2*/ΔR2 were also made on a pixel-by-pixel basis. These techniques support the hypothesis that susceptibility contrast MRI can provide useful quantitative metrics of in vivo tumor vascular morphology.     

Capacity of human monocytes to phagocytose approved iron oxide MR contrast agents in vitro

To evaluate the capacity of human monocytes to phagocytose various approved iron oxide based magnetic resonance (MR) contrast agents and to optimize in vitro labeling of these cells. Human monocytes were incubated with two superparamagnetic iron oxide particles (SPIO) as well as two ultrasmall SPIO (USPIO) at varying iron oxide concentrations and incubation times. Iron uptake in monocytes was proven by histology, quantified by atomic emission absorption spectrometry and depicted with T2* weighted fast field echo (FFE) MR images at 1.5 T. Additionally, induction of apoptosis in iron oxide labeled monocytes was determined by YO-PRO-1 staining. Cellular iron uptake was significantly (P<0.01) higher after incubation with SPIO compared with USPIO. For SPIO, the iron oxide uptake was significantly (P<0.01) higher after incubation with the ionic Ferucarbotran as compared with the non-ionic Ferumoxides. Efficient cell labeling was achieved after incubation with Ferucarbotran at concentrations 500 g Fe/ml and incubation times 1 h, resulting in a maximal iron oxide uptake of up to 50 pg Fe/cell without impairment of cell viability. In vitro labeling of human monocytes for MR imaging is most effectively obtained with the approved SPIO Ferucarbotran. Potential subsequent in vivo cell tracking applications comprise, e.g. specific targeting of inflammatory processes.     

Magnetic resonance imaging contrast agents: Overview and perspectives

Magnetic resonance imaging (MRI) is a non-invasive clinical imaging modality, which has become widely used in the diagnosis and/or staging of human diseases around the world. Some MRI examinations include the use of contrast agents. The categorizations of currently available contrast agents have been described according to their effect on the image, magnetic behavior and biodistribution in the body, respectively. In this field, superparamagnetic iron oxide particles and soluble paramagnetic metal chelates are two main classes of contrast agents for MRI. This review outlines the research and development of MRI contrast agents. In future, the ideal MRI contrast agent will be focused on the neutral tissue- or organ-targeting materials with high relaxivity and specificity, low toxicity and side effects, suitable long intravascular duration and excretion time, high contrast enhancement with low dose in vivo, and with minimal cost.     

Theory of 1/T1 and 1/T2 NMRD profiles of solutions of magnetic nanoparticles

Organically coated iron oxide crystallites with diameters of 5–50 nm (“nanoparticles”) are potential magnetic resonance imaging contrast agents. 1/T₁ and 1/T₂ of solvent water protons are increased dramatically by magnetic interactions in the “outer sphere” environment of the nanoparticles; subsequent diffusive mixing distributes this relaxation throughout the solvent. Published theory, valid for the solute magnetic energy small compared with thermal energy, is applicable to small magnetic solutes (e.g., gadolinium and manganese diethylenetriaminopentaacetic acid, and nitroxide free radicals) at generally accessible fields (≤ 50 T). It fails for nanoparticles at fields above ?0.05 T, i.e., at most imaging fields. The authors have reformulated outer sphere relaxation theory to incorporate progressive magnetic saturation of solute nanoparticles and, in addition, indicate how to use empirical magnetization data for realistic particles when their magnetic properties are not ideal. It is important to handle the effects of rapid thermally induced reorientation of the magnetization of the nanoparticles (their “superparamagnetism”) effectively, including their sensitivity to particle size. The theoretical results are presented as the magnetic field dependence (NMRD profiles) of 1/T₁ and 1/T₂, normalized to Fe content, for three sizes of particles, and then compared with the limited data extant for well-characterized material.     

A quantitative study of relaxation rate enhancement produced by iron oxide particles in polyacrylamide gels and tissue

The proton relaxation rate enhancements produced by two types of iron oxide particles, M4125 and AM125 (Advanced Magnetics, Cambridge, MA) designed as potential MR contrast agents, have been measured in polyacrylamide gels and in liver and spleen. The organ uptakes, time courses, and biodistributions of these materials have been measured using radiolabeling. The relaxation rate enhancements produced by these particles have been related to the concentration of iron per gram material at different magnetic field strengths. The relaxation rates increase linearly as a function of concentration at field strengths varying from 0.15 to 7 T. The relaxation effects of these particles in tissue are significantly different from the behavior in vitro. These results suggest that several different mechanisms contribute to relaxation effects in the presence of iron oxide particles and they depend strongly on the way the particles are sequestered. Diffusion effects for a specified TE may be enhanced using gradient or single-spin echoes rather than multiple echoes. AM125 is significantly more effective as a relaxation agent than paramagnetic metal ions only at relatively high fields, while M4125 is less effective per unit concentration.     

Superparamagnetic iron oxide nanoparticles as a dual imaging probe for targeting hepatocytes in vivo

Hepatocyte‐specific targeting agents are useful for evaluation of the hepatocytic function and the monitoring of disease progress. Superparamagnetic iron oxide nanoparticles (SPION) bearing terminal galactose groups exhibit a high affinity for the asialoglycoprotein receptor on the hepatocyte surface. In this study, we synthesized and characterized the dual probe SPION detectable by both nuclear and MR imaging modality for specifically targeting hepatocytes in vivo. SPION with 12‐nm diameter were functionalized with dopamine. Surface modification of the SPION was performed to target asialoglycoprotein receptor on hepatocytes, using lactobionic acid. Transmission electron microscope images demonstrated that SPION displayed highly uniform characteristics in terms of both particle size and shape. The X‐ray diffraction pattern of SPION revealed a nanocrystal structure of magnetite. To radiolabel the magnetite with ^99mTc, diethylenetriaminepentaacetic acid was conjugated to unreacted functional groups of dopamine. ^99mTc‐labeled lactobionic acid‐SPION showed high accumulation in liver, with 38.43 ± 6.45% injected dose per gram. In MR imaging, the reduction of the T₂ signal in the liver by lactobionic acid‐SPION was approximately 50.8 ± 7.3%. Competition studies and transmission electron microscope images of liver tissues demonstrated that the lactobionic acid‐SPION were localized in hepatocytes. Therefore, the lactobionic acid‐SPION may be used as a hepatocyte‐targeted dual contrast agent for both nuclear and MR imaging. Magn Reson Med, 2009. © 2009 Wiley‐Liss, Inc.     

NC100150 Injection, a preparation of optimized iron oxide nanoparticles for positive-contrast MR angiography

A preparation of monocrystalline iron oxide nanoparticles with an oxidized starch coating, currently in clinical trials (NC100150 Injection; CLARISCAN), was characterized by magnetization measurements, relaxometry, and photon correlation spectroscopy. By combining the results with a measure of iron content, one can obtain the size and magnetic attributes of the iron cores, including the relevant correlation times for outer sphere relaxation (tau(SO) and tau(D)), and information about the interaction of the organic coating with both core and solvent. The results are 6.43 nm for the iron oxide core diameter, a magnetic moment of 4.38x10(-17) erg/G, and a water-penetrable coating region of oxidized oligomeric starch fragments and entrained water molecules. The latter extends the hydrodynamic diameter to 11.9 nm and lowers the average diffusivity of solvent about 64% (which increases tau(D) accordingly). The nanoparticles show little size-polydispersity, evidenced by the lowest value of r(2)/r(1) at 20 MHz reported to date, an asset for magnetic resonance angiography.     

Release activation of iron oxide nanoparticles: (REACTION) A novel environmentally sensitive MRI paradigm

Smart contrast agents for MRI‐based cell tracking would enable the use of MRI methodologies to not only detect the location of cells but also gene expression. Here, we report on a new enzyme/contrast agent paradigm which involves the enzymatic degradation of the polymer coating of magnetic nanoparticles to release encapsulated magnetic cores. Cells were labeled with particles coated with a polymer, which is cleavable by a specific enzyme. This coat restricts the approach of water to the particle, preventing the magnetic core from efficiently relaxing protons. The reactive enzyme was delivered to cells and changes in cellular T₂ and T₂* relaxation times of ～ 35% and ～ 50% were achieved in vitro. Large enhancements of dark contrast volume (240%) and contrast‐to‐noise ratio (48%) within the contrast regions were measured, in vivo, for cells co‐labeled with enzyme and particles. These results warrant exploration of genetic avenues toward achieving release activation of iron oxide nanoparticles. Magn Reson Med, 2011. © 2011 Wiley‐Liss, Inc.     

In Vivo sodium multiple quantum spectroscopy of human articular cartilage

The authors report, for the first time, sodium properties of human articular cartilage in vivo using sodium multiple-quantum-filtered methods with a surface coil. A flip angle-independent, phase-cycled pulse sequence was used to obtain triple-quantum-filtered spectra as a function of preparation time. Biexponential relaxation rates were calculated by fitting the triple-quantum-filtered spectral amplitudes to a theoretical expression. Theoretical analysis of the flip angle dependence of even rank two-quantum coherence (T₂²), odd rank two-quantum coherence (T₂³), and triple-quantum coherence are presented and verified against experimental results on a cartilage specimen. Sodium multiple-quantum-filtered spectral lineshapes obtained in vivo correlate well with those observed on in vivo specimens. Relaxation rates obtained from asymptomatic volunteers were found to be: T_2rise= 1.0 ± 0.12 ms, T_2decay= 12.0 ± 0.75 ms (mean ± SD). The diagnostic potential of this method in detecting early changes in articular cartilage is described.     

Accelerated stem cell labeling with ferucarbotran and protamine

Objective

To develop and characterize a clinically applicable, fast and efficient method for stem cell labeling with ferucarbotran and protamine for depiction with clinical MRI.

Methods

The hydrodynamic diameter, zeta potential and relaxivities of ferucarbotran and varying concentrations of protamine were measured. Once the optimized ratio was found, human mesenchymal stem cells (MSCs) were labeled at varying incubation times (1–24 h). Viability was assessed via Trypan blue exclusion testing. 150,000 labeled cells in Ficoll solution were imaged with T1-, T2- and T2*-weighted sequences at 3 T, and relaxation rates were calculated.

Results

Varying the concentrations of protamine allows for easy modification of the physicochemical properties. Simple incubation with ferucarbotran alone resulted in efficient labeling after 24 h of incubation while assisted labeling with protamine resulted in similar results after only 1 h. Cell viability remained unaffected. R2 and R2* relaxation rates were drastically increased. Electron microscopy confirmed intracellular iron oxide uptake in lysosomes. Relaxation times correlated with results from ICP-AES.

Conclusion

Our results show internalization of ferucarbotran can be accelerated in MSCs with protamine, an approved heparin antagonist and potentially clinically applicable uptake-enhancing agent.     

Emerging applications for ferumoxytol as a contrast agent in MRI

Ferumoxytol is an ultrasmall superparamagnetic iron oxide (USPIO) agent initially approved by the Food and Drug Administration (FDA) as an iron replacement therapy for patients with anemia due to chronic renal failure. Recently, ferumoxytol has been investigated extensively as an intravenous contrast agent in magnetic resonance imaging (MRI). Since it causes regional T₁ and T₂* shortening in vivo, conventional pulse sequences can be used following ferumoxytol administration to demonstrate signal enhancement or loss. Ferumoxytol can be administered as a rapid bolus and has a long intravascular half‐life on the order of 14–15 hours, making it a potentially useful agent for vascular and perfusion‐weighted MRI. In comparison to other USPIOs, ferumoxytol is less limited by allergic and idiosyncratic reactions. Furthermore, since ferumoxytol is an iron‐based agent with no potential for causing nephrogenic systemic fibrosis, it may be useful as an alternative to gadolinium‐based contrast agents in patients with compromised renal function. Ferumoxytol is ultimately taken up by macrophages/the reticuloendothelial system in the liver, spleen, and lymph nodes, and this uptake mechanism is being explored as a novel imaging technique for vascular lesions, tumors, and lymph nodes. This article reviews the properties of ferumoxytol relevant to MRI as well as many of the uses for the agent currently under investigation. J. Magn. Reson. Imaging 2015;41:884–898 . © 2014 Wiley Periodicals, Inc .