Effect of diffusion time on liver DWI: an experimental study of normal and fibrotic livers.

PURPOSE: To investigate whether diffusion time (Δ) affects the diffusion measurements in liver and their sensitivity in detecting fibrosis. METHODS: Liver fibrosis was induced in Sprague-Dawley rats (n = 12) by carbon tetrachloride (CCl(4)) injections. Diffusion-weighted MRI was performed longitudinally during 8-week CCl(4) administration at 7 Tesla (T) using single-shot stimulated-echo EPI with five b-values (0 to 1000 s/mm(2)) and three Δs. Apparent diffusion coefficient (ADC) and true diffusion coefficient (D(true)) were calculated by using all five b-values and large b-values, respectively. RESULTS: ADC and D(true) decreased with Δ for both normal and fibrotic liver at each time point. ADC and D(true) also generally decreased with the time after CCl(4) insult. The reductions in D(true) between 2-week and 4-week CCl(4) insult were larger than the ADC reductions at all Δs. At each time point, D(true) measured with long Δ (200 ms) detected the largest changes among the 3 Δs examined. Histology revealed gradual collagen deposition and presence of intracellular fat vacuoles after CCl(4) insult. CONCLUSION: Our results demonstrated the Δ dependent diffusion measurements, indicating restricted diffusion in both normal and fibrotic liver. D(true) measured with long Δ acted as a more sensitive index of the pathological alterations in liver microstructure during fibrogenesis.

Molecular MRI of liver fibrosis by a peptide-targeted contrast agent in an experimental mouse model.

OBJECTIVES: Cyclic decapeptide CGLIIQKNEC (CLT1) has been demonstrated to target fibronectin-fibrin complexes in the extracellular matrix of different tumors and tissue lesions. Although liver fibrosis is characterized by an increased amount of extracellular matrix consisting of fibril-forming collagens and matrix glycoconjugates such as fibronectin, we aimed to investigate the feasibility of detecting and characterizing liver fibrosis using CLT1 peptide-targeted nanoglobular contrast agent (Gd-P) with dynamic contrast-enhanced magnetic resonance imaging in an experimental mouse model of liver fibrosis at 7 T. MATERIALS AND METHODS: Gd-P, control peptide KAREC conjugated nanoglobular contrast agent (Gd-CP), and control nontargeting nanoglobular contrast agent (Gd-C) were synthesized. Male adult C57BL/6N mice (22-25 g; N = 54) were prepared and were divided into fibrosis (n = 36) and normal (n = 18) groups. Liver fibrosis was induced in the fibrosis group through subcutaneous injection of 1:3 mixture of carbon tetrachloride (CCl(4)) in olive oil at a dose of 4 µL/g of body weight twice a week for 8 weeks. Dynamic contrast-enhanced MRI was performed in all animals. Dynamic contrast-enhanced magnetic resonance imaging was analyzed to yield postinjection ΔR(1)(t) maps for quantitative measurements. Histological analysis was also performed. RESULTS: Differential enhancements were observed and characterized between the normal and fibrotic livers using Gd-P at 0.03 mmol/kg, when compared with nontargeted controls (Gd-CP and Gd-C). For Gd-P injection, both the peak and steady-state ΔR(1) of the normal livers were significantly lower than those after 4 and 8 weeks of CCl(4) dosing. Liver fibrogenesis with increased amount of fibronectin in the extracellular space in insulted livers were confirmed by histological observations. CONCLUSIONS: These results indicated that dynamic contrast-enhanced magnetic resonance imaging with CLT1 peptide-targeted nanoglobular contrast agent can detect and stage liver fibrosis by probing the accumulation of fibronectin in fibrotic livers.

Measurement of liver T1 and T2 relaxation times in an experimental mouse model of liver fibrosis.

PURPOSE: To characterize changes in relaxation times of liver using quantitative magnetic resonance imaging (MRI) in an experimental mouse model of liver fibrosis. Quantitative MRI is a potentially robust method to characterize liver fibrosis. However, correlation between relaxation times and fibrosis stage has been controversial. MATERIALS AND METHODS: Liver fibrosis was induced in male adult C57BL/6N mice (22-25 g; n = 12) by repetitive dosing of carbon tetrachloride (CCl(4) ). The animals were examined with a series of spin-echo (SE) images with varying TRs and multiecho SE imaging sequence at 7 T before and 2, 4, 6, and 8 weeks after CCl(4) insult. Hepatic T(1) and T(2) values were measured. Histology was performed with hematoxylin-eosin staining and Masson's trichrome staining. RESULTS: Significant increase (P < 0.001) in hepatic T(1) was found at 2, 4, 6, and 8 weeks following CCl(4) insult as compared with that before insult. Meanwhile, hepatic T(2) at 2, 4, 6, and 8 weeks after CCl(4) insult was significantly higher (P < 0.001) than that before the insult. Liver histology showed collagen deposition, edema, and infiltration of inflammatory cells in livers with CCl(4) insult. CONCLUSION: Both longitudinal and transverse relaxation times may serve as robust markers for liver fibrosis. With the advent of single breath-hold sequences for MR relaxometry, quantitative mapping of relaxation times can be routinely and reliably performed in abdominal organs and hence may be valuable and robust in detecting liver fibrosis at early phase and monitoring its progression.

Liver fibrosis: an intravoxel incoherent motion (IVIM) study.

PURPOSE: To characterize longitudinal changes in molecular water diffusion, blood microcirculation, and their contributions to the apparent diffusion changes using intravoxel incoherent motion (IVIM) analysis in an experimental mouse model of liver fibrosis. MATERIALS AND METHODS: Liver fibrosis was induced in male adult C57BL/6N mice (22-25 g; n = 12) by repetitive dosing of carbon tetrachloride (CCl(4) ). The respiratory-gated diffusion-weighted (DW) images were acquired using single-shot spin-echo EPI (SE-EPI) with 8 b-values and single diffusion gradient direction. True diffusion coefficient (D(true) ), blood pseudodiffusion coefficient (D(pseudo) ), and perfusion fraction (P(fraction) ) were measured. Diffusion tensor imaging (DTI) was also performed for comparison. Histology was performed with hematoxylin-eosin and Masson's trichrome staining. RESULTS: A significant decrease in D(true) was found at 2 weeks and 4 weeks following CCl(4) insult, as compared with that before insult. Similarly, D(pseudo) values before injury was significantly higher than those at 2 weeks and 4 weeks after CCl(4) insult. Meanwhile, P(fraction) values showed no significant differences over different timepoints. For DTI, significant decrease in ADC was observed following CCl(4) administration. Fractional anisotropy at 2 weeks after CCl(4) insult was significantly lower than that before insult, and subsequently normalized at 4 weeks after the insult. Liver histology showed collagen deposition, the presence of intracellular fat vacuoles, and cell necrosis/apoptosis in livers with CCl(4) insult. CONCLUSION: Both molecular water diffusion and blood microcirculation contribute to the alteration in apparent diffusion changes in liver fibrosis. Reduction in D(true) and D(pseudo) values resulted from diffusion and perfusion changes, respectively, during the progression of liver fibrosis. IVIM analysis may serve as valuable and robust tool in detecting and characterizing liver fibrosis at early stages, monitoring its progression in a noninvasive manner.

Metabolic changes in visual cortex of neonatal monocular enucleated rat: a proton magnetic resonance spectroscopy study.

Neonatal monocular enucleation (ME) is often employed to study the developmental mechanisms underlying visual perception and the cross-modal changes in the central nervous system caused by early loss of the visual input. However, underlying biochemical or metabolic mechanisms that accompany the morphological, physiological and behavioral changes after ME are not fully understood. Male Sprague-Dawley rats (N=14) were prepared and divided into 2 groups. The enucleated group (N=8) underwent right ME (right eye removal) at postnatal day 10, while the normal group (N=6) was intact and served as a control. Three weeks after ME, single voxel proton magnetic resonance spectroscopy ((1)H MRS) was performed over the visual cortex of each hemisphere in all animals with a point-resolved spectroscopy (PRESS) sequence at 7 T. The taurine (Tau) and N-acetylaspartate (NAA) levels were found to be significantly lower in the left visual cortex (contralateral to enucleated eye) for enucleated animals. Such metabolic changes measured in vivo likely reflected the cortical degeneration associated with the reduction of neurons, axon terminals and overall neuronal activity. This study also demonstrated that (1)H MRS approach has the potential to characterize neonatal ME and other developmental neuroplasticity models noninvasively for the biochemical and metabolic processes involved.

In vivo lipid profiling using proton magnetic resonance spectroscopy in an experimental liver fibrosis model.

RATIONALE AND OBJECTIVES: The aim of this study was to characterize early hepatic lipid changes in an experimental model of liver fibrosis using proton ((1)H) magnetic resonance spectroscopy (MRS) at high magnetic field in vivo. MATERIALS AND METHODS: Liver fibrosis was induced in 12 Sprague-Dawley rats by twice-weekly carbon tetrachloride (CCl(4)) administration up to 4 weeks. Eight normal rats were used as controls. Single-voxel (1)H MRS experiments were performed at 7 Tesla to measure signal integrals of various lipid peaks including -CH(3), (-CH(2)-)(n), -CH(2)-C=C-CH(2)-, =C-CH(2)-C= and -CH=CH- at 0.9, 1.3, 2.0, 2.8, and 5.3 ppm, respectively, and peak from choline-containing compounds (CCC) at 3.2 ppm. Total lipid, total saturated fatty acid, total unsaturated fatty acid, total unsaturated bond, polyunsaturated bond, and CCC indices were quantified. RESULTS: Significant increases (P < .01) in total lipid and total saturated fatty acid indices were found in animals with CCl(4)-induced fibrosis as compared with normal animals. In addition, total unsaturated bond and polyunsaturated bond indices of animals at 4 weeks after CCl(4) insult were significantly higher than (P < .01 and P < .05, respectively) those of normal animals and animals at 2 weeks following insult; whereas there was only significant increase (P < .01) in total unsaturated fatty acid index in animals with 4-week CCl(4) insult as compared with normal animals. CONCLUSION: The hepatic lipid changes in CCl(4)-induced experimental fibrosis model were documented in vivo and longitudinally using (1)H MRS at 7 Tesla. The experimental findings suggested that total saturated fatty acid increase contributed mainly to the total lipid increase in animals with CCl(4) insult. This study also demonstrated the potential value of high field MRS to resolve lipid composition and alterations in liver fibrosis.

In vivo proton magnetic resonance spectroscopy of hepatic ischemia/reperfusion injury in an experimental model.

RATIONALE AND OBJECTIVES: Hepatic ischemia/reperfusion injury (IRI) occurs during certain hepatobiliary surgeries, hemorrhagic shock, and veno-occlusive disease. Biochemical changes caused by hepatic IRI lead to hepatocellular remodeling, including cellular regeneration or irreversible apoptosis. This study aims to characterize and monitor the metabolic changes in hepatic IRI using proton magnetic resonance spectroscopy (¹H MRS). MATERIALS AND METHODS: Sprague-Dawley rats (n = 8) were scanned with ¹H MRS using 5.0 × 5.0 × 5.0 mm³ voxel over a homogeneous liver parenchyma at 7 Tesla with a respiratory-gated point-resolved spectroscopy sequence at 1 day before, 6 hours, 1 day, and 1 week after 30 minutes total hepatic IRI. Signal integral ratios of choline-containing compounds (CCC), glycogen and glucose complex (Glyu), methylene proton ((-CH2-)(n)), and methene proton (-CH=CH-) to lipid (integral sum of methyl proton (-CH3), (-CH2-)(n) and -CH=CH-) were quantified by areas under peaks longitudinally. RESULTS: The CCC-to-lipid and Glyu-to-lipid ratios at 6 hours after IRI were significantly higher than those at 1 day before, 1 day, and 1 week after injury. The (-CH2-)(n)-to-lipid, and -CH=CH-to-lipid ratios showed no significant differences over different time points. Hepatocellular regeneration was observed at 6 hours after IRI in histology with immunohistochemical technique. CONCLUSIONS: Changes in CCC-to-lipid and Glyu-to-lipid ratios likely reflect the hepatocellular remodeling and impaired glucose utilization upon hepatic IRI, respectively. The experimental findings in the current study demonstrated that ¹H MRS is a valuable tool for characterizing either global or regional metabolic changes in liver noninvasively and longitudinally. Such capability has the potential to lead to early diagnosis and detection of impaired liver function.

In vivo MRI study of the visual system in normal, developing and injured rodent brains.

This paper demonstrated our recent use of contrast-enhanced MRI, diffusion tensor/kurtosis imaging, proton magnetic resonance spectroscopy, and functional MRI techniques, for in vivo and global assessments of the structure, metabolism and function of the visual system in rodent studies of ocular diseases, optic neuropathies, developmental plasticity and neonatal hypoxic-ischemic brain injury at 7T. Results suggested the significant values of high-field multiparametric MRI for uncovering the processes and mechanisms of developmental and pathophysiological changes systematically along both anterior and posterior visual pathways, and may provide early diagnoses and therapeutic strategies for promoting functional recovery upon partial vision loss.

Diffusion tensor imaging of liver fibrosis in an experimental model.

PURPOSE: To characterize changes in diffusion properties of liver using diffusion tensor imaging (DTI) in an experimental model of liver fibrosis. MATERIALS AND METHODS: Liver fibrosis was induced in Sprague-Dawley rats (n = 12) by repetitive dosing of carbon tetrachloride (CCl(4)). The animals were examined with a respiratory-gated single-shot spin-echo echo-planar DTI protocol at 7 T before, 2 weeks after, and 4 weeks after CCl(4) insult. Apparent diffusion coefficient (ADC), directional diffusivities (ADC(//) and ADC(⊥)), and fractional anisotropy (FA) were measured. Liver histology was performed with hematoxylin-eosin staining and Masson's trichrome staining. RESULTS: Significant decrease (P < 0.01) in ADC was found at 2 weeks (0.86 ± 0.09 × 10(-3) mm(2)/s) and 4 weeks (0.74 ± 0.09 × 10(-3) mm(2)/s) following CCl(4) insult, as compared with that before insult (0.97 ± 0.08 × 10(-3) mm(2)/s). Meanwhile, FA at 2 weeks (0.18 ± 0.03) after CCl(4) insult was significantly lower (P < 0.01) than that before insult (0.26 ± 0.05), and subsequently normalized at 4 weeks (0.26 ± 0.07) after the insult. Histology showed collagen deposition, presence of intracellular fat vacuoles, and cell necrosis/apoptosis in livers with CCl(4) insult. CONCLUSION: DTI detected the progressive changes in water diffusivities and diffusion anisotropy of liver tissue in this liver fibrosis model. ADC and FA are potentially valuable in detecting liver fibrosis at early stages and monitoring its progression. Future human studies are warranted to further verify the applicability of DTI in characterizing liver fibrosis and to determine its role in clinical settings.

Magnetic resonance spectroscopy of the brain under mild hypothermia indicates changes in neuroprotection-related metabolites.

Brain hypothermia has demonstrated pronounced neuroprotective effect in patients with cardiac arrest, ischemia and acute liver failure. However, its underlying neuroprotective mechanisms remain to be elucidated in order to improve therapeutic outcomes. Single voxel proton magnetic resonance spectroscopy ((1)H-MRS) was performed using a 7 Tesla MRI scanner on normal Sprague-Dawley rats (N=8) in the same voxel under normothermia (36.5 degrees C) and 30min mild hypothermia (33.5 degrees C). Levels of various brain proton metabolites were compared. The level of lactate (Lac) and myo-inositol (mI) increased in the cortex during hypothermia. In the thalamus, taurine (Tau), a cryogen in brain, increased and choline (Cho) decreased. These metabolic alterations indicated the onset of a number of neuroprotective processes that include attenuation of energy metabolism, excitotoxic pathways, brain osmolytes and thermoregulation, thus protecting neuronal cells from damage. These experimental findings demonstrated that (1)H-MRS can be applied to investigate the changes of specific metabolites and corresponding neuroprotection mechanisms in vivo noninvasively, and ultimately improve our basic understanding of hypothermia and ability to optimize its therapeutic efficacy.

Diffusion tensor imaging of renal ischemia reperfusion injury in an experimental model.

Renal ischemia reperfusion injury (IRI) is a major cause of acute renal failure. It occurs in various clinical settings such as renal transplantation, shock and vascular surgery. Serum creatinine level has been used as an index for estimating the degree of renal functional loss in renal IRI. However, it only evaluates the global renal function. In this study, diffusion tensor imaging (DTI) was used to characterize renal IRI in an experimental rat model. Spin-echo echo-planar DTI with b-value of 300 s/mm(2) and 6 diffusion gradient directions was performed at 7 T in 8 Sprague-Dawley (SD) with 60-min unilateral renal IRI and 8 normal SD rats. Apparent diffusion coefficient (ADC), directional diffusivities and fractional anisotropy (FA) were measured at the acute stage of IRI. The IR-injured animals were also examined by diffusion-weighted imaging with 7 b-values up to 1000 s/mm(2) to estimate true diffusion coefficient (D(true)) and perfusion fraction (P(fraction)) using a bi-compartmental model. ADC of injured renal cortex (1.69 +/- 0.24 x 10(-3) mm(2)/s) was significantly lower (p < 0.01) than that of contralateral intact cortex (2.03 +/- 0.35 x 10(-3) mm(2)/s). Meanwhile, both ADC and FA of IR-injured medulla (1.37 +/- 0.27 x 10(-3) mm(2)/s and 0.28 +/- 0.04, respectively) were significantly less (p < 0.01) than those of contralateral intact medulla (2.01 +/- 0.38 x 10(-3) mm(2)/s and 0.36 +/- 0.04, respectively). The bi-compartmental model analysis revealed the decrease in D(true) and P(fraction) in the IR-injured kidneys. Kidney histology showed widespread cell swelling and erythrocyte congestion in both cortex and medulla, and cell necrosis/apoptosis and cast formation in medulla. These experimental findings demonstrated that DTI can probe both structural and functional information of kidneys following renal IRI.

Enhancement of gas-filled microbubble R2* by iron oxide nanoparticles for MRI.

Gas-filled microbubbles have the potential to become a unique intravascular MR contrast agent due to their magnetic susceptibility effect, biocompatibility, and localized manipulation via ultrasound cavitation. However, microbubble susceptibility effect is relatively weak when compared with other intravascular MR susceptibility contrast agents. In this study, enhancement of microbubble susceptibility effect by entrapping monocrystalline iron oxide nanoparticles (MIONs) into polymeric microbubbles was investigated at 7 T in vitro. Apparent T2 enhancement (DeltaR2*) induced by microbubbles was measured to be 79.2+/-17.5 sec(-1) and 301.2+/-16.8 sec(-1) for MION-free and MION-entrapped polymeric microbubbles at 5% volume fraction, respectively. DeltaR2* and apparent transverse relaxivities (r2*) for MION-entrapped polymeric microbubbles and MION-entrapped solid microspheres (without gas core) were also compared, showing the synergistic effect of the gas core with MIONs. This is the first experimental demonstration of microbubble susceptibility enhancement for MRI application. This study indicates that gas-filled polymeric microbubble susceptibility effect can be substantially increased by incorporating iron oxide nanoparticles into microbubble shells. With such an approach, microbubbles can potentially be visualized with higher sensitivity and lower concentrations by MRI.

Gas-filled microbubbles--a novel susceptibility contrast agent for brain and liver MRI.

Gas-filled microbubbles have the potential to become a unique intravascular MR contrast agent due to their magnetic susceptibility effect, biocompatibility and localized manipulation via ultrasound cavitation. However, in vivo demonstration of microbubble susceptibility effect is limited so far and microbubble susceptibility effect is relatively weak when compared with other intravascular MR susceptibility contrast agents. In this study, two types of microbubbles, custom-made albumin-coated microbubbles (AMBs) and a commercially available lipid-based clinical ultrasound contrast agent (SonoVue), were investigated with in vivo dynamic brain and liver MRI in Sprague-Dawley rats at 7 Tesla. Transverse relaxation rate enhancements (DeltaR2*) maps were computed for brain and liver, yielding results similar to those obtained with a common MR blood pool contrast agent. These results indicate that gas-filled microbubbles can serve as an intravascular MR contrast agent at high field. Enhancement of microbubble susceptibility effect by entrapping monocrystalline iron oxide nanoparticles (MIONs) into microbubbles was also investigated at 7 T in vitro. This is the first experimental demonstration of microbubble susceptibility enhancement for MRI application. This study indicates that gas-filled microbubble susceptibility effect can be substantially increased using iron oxides nanoparticles. With such approach, microbubbles can potentially be visualized with higher sensitivity and lower concentrations by MRI. Such capability has the potential to lead to real-time MRI guidance in various microbubble-based drug delivery and therapeutic applications.

In vivo DTI assessment of hepatic ischemia reperfusion injury in an experimental rat model.

PURPOSE: To investigate hepatic ischemia reperfusion injury (IRI) using diffusion tensor imaging (DTI). MATERIALS AND METHODS: Ten Sprague-Dawley rats were scanned at 7 Tesla (T) with DTI using b-value of 1000 s/mm(2) and 6 gradient directions before, 2 h, and 1 day after 30-min total hepatic IRI. Apparent diffusion coefficient or mean diffusivity (MD), directional diffusivities and fractional anisotropy (FA) were measured. Seven of the animals were also examined with spin-echo echo-planar diffusion-weighted imaging (DWI) with seven b-values up to 2000 s/mm(2) to estimate the true diffusion coefficient (D), blood pseudodiffusion coefficient (D), and perfusion fraction (f) using a bi-compartmental model. RESULTS: MD 2 h after IRI (0.77 +/- 0.07 x 10(-3) mm(2)/s) was significantly lower (P < 0.01) than that before (1.03 +/- 0.07 x 10(-3) mm(2)/s) and 1 day after IRI (1.01 +/- 0.05 x 10(-3) mm(2)/s). Meanwhile, FA 2 h after IRI (0.33 +/- 0.03) was significantly higher (P < 0.01) than that before (0.21 +/- 0.02) and 1 day after IRI (0.20 +/- 0.02). The bi-compartmental model analysis revealed the transient decrease in D, D and f 2 h after IRI. Liver histology showed the multifocal cell swelling 3 h after IRI and widespread cell necrosis/apoptosis 1 day after IRI. Sinusoidal narrowing and congestion of erythrocytes were also observed 3 h and 1 day after IRI. CONCLUSION: DTI can characterize hepatic IRI by detecting the transient change in both MD and FA.

Microbubbles as a novel contrast agent for brain MRI.

Gas-filled microbubbles have the potential to become a unique MR contrast agent due to their magnetic susceptibility effect, biocompatibility and localized manipulation via ultrasound cavitation. In this study, two types of microbubbles, custom-made albumin-coated microbubbles (A-MB) and a commercially available lipid-based clinical ultrasound contrast agent (SonoVue), were investigated with in vivo dynamic brain MRI in Sprague-Dawley rats at 7 T. Microbubble suspensions (A-MB: 0.2 mL of approximately 4% volume fraction; SonoVue: 0.2 mL of approximately 3.5% volume fraction) were injected intravenously. Transverse relaxation rate enhancements (DeltaR(2)(*)) of 2.49+/-1.00 s(-1) for A-MB and 2.41+/-1.18 s(-1) for SonoVue were observed in the brain (N=5). Brain DeltaR(2)(*) maps were computed, yielding results similar to the cerebral blood volume maps obtained with a common MR blood pool contrast agent. Microbubble suspension DeltaR(2)(*) was measured for different volume fractions. These results indicate that gas-filled microbubbles can serve as an intravascular contrast agent for brain MRI at high field. Such capability has the potential to lead to real-time MRI guidance in various microbubble-based drug delivery and therapeutic applications in the central nervous system.

Cell number quantification of USPIO-labeled stem cells by MRI: an in vitro study.

MRI plays an expanding role in stem cell therapies. The non-invasive nature and high spatial resolution of MR imaging make MR imaging a powerful tool to investigate biologic processes at the molecular and cellular level in vivo longitudinally. Quantitative detection of stem cells after transplantation may allow assessment of stem cell localization and migration, and monitoring of the therapeutic effectiveness of stem cell therapy. In this study, we present a technique for MR quantification of magnetically labeled mouse embryonic stem cells distributed or injected in agarose gel phantoms. Apparent transverse relaxation rate enhancements (DeltaR2*) were measured by gradient echo sequences. The linear relationship between DeltaR2* and the concentration of USPIO-labeled mouse embryonic stem cells was observed and used for quantifying cell density and cell number after injection or transplantation. The MRI acquisition and analysis protocol were validated by good agreement between actual cell numbers and MRI-estimated cell numbers over a wide range of cell numbers. This MR technique for cell number and cell density quantification is applicable to future in vivo studies.