Decrease In T2 Relaxation Times Research Articles

Experimental Study on the Impact of Clay Distribution and Mineralogy on NMR T2 and Internal Gradient: Insights From Well-Controlled Physical Samples

Summary Nuclear magnetic resonance (NMR) is a reliable tool for petrophysical evaluation and the characterization of pore structures. Compared to conventional carbonate reservoirs, sandstone reservoirs contain higher amounts of ferromagnetic and paramagnetic ions (such as iron, nickel, or manganese) usually found in microporous clay aggregates. The interpretation of petrophysical data in sandstone formations can be complicated by variations in clay mineralogy and distribution patterns (laminated, structural, and dispersed). Nevertheless, the impact of clay distribution patterns on NMR signals is not well understood. This study aims to investigate the impact of clay mineralogy and distribution patterns on the T2 relaxation times and internal gradient (i.e., inhomogeneity in the magnetic field). Glass beads were mixed with three different clay minerals characterized by no iron content (kaolinite) to higher iron content (illite and nontronite). The bead-clay mixtures were prepared at a fixed clay content but with variable clay distribution patterns to examine the impact of clay distribution alone. NMR T2 measurements at several echo times were performed on the pure glass beads and the mixtures to evaluate how clay mineralogy and distribution patterns affect the T2 and the internal gradient of the host glass beads. At a given clay distribution pattern, a more significant decrease in T2 relaxation times and a larger increase in the internal gradient of glass beads were observed when adding clays with higher iron content. This is explained by the higher surface relaxivity, and magnetic susceptibility caused when introducing clay with higher iron content. Such an impact can complicate the characterization of NMR-derived pore sizes as similar pore size distribution (PSD) can have very different T2 distribution and the logarithmic mean of T2 relaxation time distribution (T2LM) values. Micro-computed tomography (µCT) images were acquired to compute the PSD to compare it with ones obtained from the NMR measurements. The PSD for the three clay minerals showed almost the same distribution using µCT; however, they showed totally different T2 relaxation times distributions. That is due to the significant impact of the internal gradient causing a distortion in the magnetic field. Thus, careful consideration must be taken before converting the NMR data into PSD. The introduction of iron-free kaolinite resulted in a negligible impact on the internal gradient of glass beads regardless of the clay distribution pattern. On the other hand, the addition of dispersed iron-rich clays (illite and nontronite) results in up to two orders of magnitude increase in internal gradients, compared to mixtures with laminated and structural clays (at the same clay mineralogy and content). Moreover, dispersed clay mixtures display larger changes in T2LM and porosity as a function of increasing echo time. The results from this study suggest that changes in T2LM and the logarithmic mean of the effective internal gradient distribution geff,LM, compared to the clean host sand, can provide insight into iron-rich clay distribution. Larger changes in any given clay content and mineralogy would suggest a more dominant dispersed clay distribution while negligible changes would suggest a laminated distribution.

The effect of weight loss on hypothalamus structure and function in obese individuals: a systematic review and meta-analysis

Introduction: Obesity presents with structural and functional hypothalamic dysfunction. However, it is unclear whether weight loss can lead to hypothalamic changes. We therefore aimed to conduct a systematic review and meta-analysis to determine the effect of body mass reduction in obese individuals on hypothalamic structure and function. Methods: PubMed, Embase and Cochrane databases were searched for studies that reported the change in hypothalamic structure and function after weight loss. Qualitative and quantitative analyses were performed on magnetic resonance imaging techniques, medio-basal hypothalamus T2-relaxation time, blood oxygen level dependent (BOLD) contrast, voxel-based morphometry (VBM) and biomarkers including glucose, insulin, leptin, ghrelin and inflammatory markers of interleukins. Mean differences between pre- and post-weight loss and 95% confidence intervals (CIs) were pooled using random-effects models. Results: Thirteen pre–post studies were included, of which six accounted for the meta-analysis. Studies showed a favorable decrease in T2-relaxation time (n = 1), favorable change in hypothalamic activity after weight loss on BOLD contrast (n = 4), with higher peak activities after surgical weight loss (n = 2). No differences were found in the gray matter density of the hypothalamus on VBM (n = 1). Pooled mean differences between pre- and post-surgical weight loss revealed a decrease of 8.53 mg/dl (95% CI: 5.17, 11.9) in glucose, 7.73 pmol/l (95% CI: 5.07, 10.4) in insulin, 15.5 ng/ml (95% CI: 9.40, 21.6) in leptin, 142.9 pg/ml (95% CI: 79.0, 206.8) in ghrelin and 9.43 pg/ml (95% CI: −6.89, 25.7) in IL-6 level. Conclusions: Our study showed weight reduction in obesity led to limited structural change and significant functional changes in the hypothalamus.

Lumbar vertebral T2-relaxation time investigated with T2-mapping at multiple time points in a day demonstrate large individual variations.

PURPOSE The increasing interest of endplate and Modic changes as potential pain generators in low back pain (LBP), along with advancement of functional quantitative magnetic resonance imaging (MRI) techniques, makes it important to characterize the vertebral dynamic behavior in detail. This study aims to perform characterization of the dynamic behavior of the vertebral bodies (VB) by investigating the VB diurnal variation in T2-relaxation time in a cross-sectional asymptomatic group of individuals. METHODS T2-mapping of 30 VBs (L1-L5) in six healthy volunteers (mean age, 40 years; range, 29-65 years) was performed with a 1.5 Tesla MRI at three time points over the day (7 am, 12 am, 5 pm). Volumetric regions of interest were segmented manually to determine VB T2-relaxation time, which was compared between the three time points. RESULTS On a group level only small and not significant diurnal VB variation was detected (all P >.10), with median T2 (ms) (quartiles; Q1, Q3) at the three time points 88.7 (84.1, 99.1), 87.3 (85.0, 96.1) and 87.8 (84.4, 99.2). However, in some VBs up to 7% increase respectively 9% decrease in T2-relaxation time was found during the day. Further, there was a relatively large variation between the individuals in absolute VB T2-relaxation times (range 73.2-108.3 ms), but small differences between the VBs within an individual. CONCLUSION This first T2-mapping study of the VB signal dynamics, in repeated investigations during one day, display variation in T2-relaxation time in specific individual VBs but were negligible on a group level. The result may be of importance when evaluating patients with spinal pathologies and suggest further examinations of dynamic changes not only of the disc but also vertebrae.

Association of changes in the infarct and remote zone myocardial tissue with cardiac remodeling after myocardial infarction: a T1 and T2 mapping study.

Tissue structure in the infarct and remote zone myocardium post-acute myocardial infarction (MI) may offer prognostic information concerning left ventricular remodeling. We aimed to identify or establish a relationship between adverse remodeling (AR) and changes (Δ) in T1, T2 mapping and extracellular volume (ECV) in post MI periods. Fifty-four MI patients underwent 3 Tesla CMR performed 2weeks (acute phase) and 6months post-MI. We measured T1 mapping with MOLLI sequences and T2 mapping with TrueFISP sequences. Hematocrit was quantified in scanning time. ECV was performed post-gadolinium enhancement. AR was defined as an increase of ≥ 10% in left ventricular end-diastolic volume in 6months. In the acute phase post-MI, high T2 relaxation times of the infarct and remote zone myocardium were associated with AR (OR 1.15, p = 0.023 and OR 1.54, p = 0.002, respectively). There was a decrease in T2 relaxation times of the remote zone myocardium at 6months in patients with AR (42.0 ± 4.0 vs. 39.0 ± 3.5ms, p < 0.001), while insignificant difference was found in patients without AR. Increased ΔECV (%) and decreased remote ΔT2 values were associated with AR (OR 1.04, p = 0.043 and OR 0.77, p = 0.007, respectively). The diagnostic performance analysis in predicting AR showed that acute-phase remote T2 was similar to that of remote ΔT2 (p = 0.875) but was superior to that of ΔECV (%) (ΔAUC: 0.19 ± 0.09, p = 0.038). In both acute phase and change of 6months post-MI, the T2 relaxation times in remote myocardium are independently associated with AR, and this suggests higher inflammation in the remote myocardium in the AR group than the other group, even though no significant pathophysiological difference was observed in the healing of the infarct zone between both groups.

Quantification of myocardial fibrosis using noninvasive T2-mapping magnetic resonance imaging: Preclinical models of aging and pressure overload.

Noninvasive imaging of cardiac fibrosis is important for early diagnosis and intervention in chronic heart diseases. Here, we investigated whether noninvasive, contrast agent-free MRI T2 -mapping can quantify myocardial fibrosis in preclinical models of aging and pressure overload. Myocardial fibrosis and remodeling were analyzed in two animal models: (i) aging (15-month-old male CF-1 mice vs. young 6- to 8-week-old mice), and (ii) pressure overload (PO; by transverse aortic constriction in 4- to 5-month-old male C57BL/6 mice vs. sham-operated for 14 days). In vivo T2 -mapping was performed by acquiring data during the isovolumic and early diastolic phases, with a modified respiratory and ECG-triggered multiecho TurboRARE sequence on a 7-T MRI. Cine MRI provided cardiac morphology and function. A quantitative segmentation method was developed to analyze the in vivo T2 -maps of hearts at midventricle, apex, and basal regions. The cardiac fibrosis area was analyzed ex vivo by picro sirius red (PSR) staining. Both aged and pressure-overloaded hearts developed significant myocardial contractile dysfunction, cardiac hypertrophy, and interstitial fibrosis. The aged mice had two phenotypes, fibrotic and mild-fibrotic. Notably, the aged fibrotic subgroup and the PO mice showed a marked decrease in T2 relaxation times (25.3 ± 0.6in aged vs. 29.9 ± 0.7ms in young mice, p = 0.002; and 24.3 ± 1.7in PO vs. 28.7 ± 0.7ms in shams, p = 0.05). However, no significant difference in T2 was detected between the aged mild-fibrotic subgroup and the young mice. Accordingly, an inverse correlation between myocardial fibrosis percentage (FP) and T2 relaxation time was derived (R2 = 0.98): T2 (ms)=30.45 - 1.05× FP. Thus, these results demonstrate a statistical agreement between T2 -map-quantified fibrosis and PSR staining in two different clinically relevant animal models. In conclusion, T2 -mapping MRI is a promising noninvasive contrast agent-free quantitative technique to characterize myocardial fibrosis.

Association between spinal alignment and biochemical composition of lumbar intervertebral discs assessed by quantitative magnetic resonance imaging.

To evaluate potential associations between spinopelvic parameters and the biochemical composition of lumbar intervertebral discs using quantitative magnetic resonance imaging in asymptomatic young adults. Our study group comprised 93 asymptomatic volunteers aged 20-40 years (49 women and 44 men). Lumbar spine T2-weighted images and T2 relaxometry were acquired on a 1.5T MRI scanner. Spinopelvic parameters including sacral slope, pelvic tilt, pelvic incidence, lumbar lordosis, thoracic kyphosis, thoracolumbar alignment, sagittal vertical axis, spinosacral angle, C2 pelvic angle, and T1S1 and L1S1 length were measured on panoramic spine radiographs. Lumbar lordosis decrease correlates with discrete dehydration of nucleus pulposus at all lumbar levels. Also low values of sacral slope, pelvic tilt, pelvic incidence, thoracic kyphosis and spinosacral angle were associated with decrease of T2 relaxation times on annulus fibrosus. In conclusion, spinopelvic parameters presented a discrete association with lumbar disc composition and water content.

Increased tumour burden alters skeletal muscle properties in the KPC mouse model of pancreatic cancer.

Cancer cachexia is a multifactorial wasting syndrome that is characterized by the loss of skeletal muscle mass and weakness, which compromises physical function, reduces quality of life, and ultimately can lead to mortality. Experimental models of cancer cachexia have recapitulated this skeletal muscle atrophy and consequent decline in muscle force generating capacity. We address these issues in a novel transgenic mouse model Kras, Trp53 and Pdx-1-Cre (KPC) of pancreatic ductal adenocarcinoma (PDA) using multi-parametric magnetic resonance (mp-MR) measures. KPC mice (n = 10) were divided equally into two groups (n = 5/group) depending on the size of the tumor i.e. tumor size <250 mm3 and >250 mm3. Using mp-MR measures, we demonstrated the changes in the gastrocnemius muscle at the microstructural level. In addition, we evaluated skeletal muscle contractile function in KPC mice using an in vivo approach. Increase in tumor size resulted in decrease in gastrocnemius maximum cross sectional area, decrease in T2 relaxation time, increase in magnetization transfer ratio, decrease in mean diffusivity, and decrease in radial diffusivity of water across the muscle fibers. Finally, we detected significant decrease in absolute and specific force production of gastrocnemius muscle with increase in tumor size. Our findings indicate that increase in tumor size may cause alterations in structural and functional parameters of skeletal muscles and that MR parameters may be used as sensitive biomarkers to noninvasively detect structural changes in cachectic muscles.

Biochemical changes in knee articular cartilage of novice half-marathon runners.

ObjectiveTo evaluate changes in knee articular cartilage of novice half-marathon runners using magnetic resonance imaging T2 relaxation time mapping.MethodsHealthy subjects were recruited from local running clubs who met the following inclusion criteria: (i) age 18–45 years; (ii) body mass index less than 30 kg/m2; (iii) had participated in one half-marathon or less (none within the previous 6 months); (iv) run less than 20 km/week; (v) no previous knee injury or surgery; (vi) no knee pain. T2 signals were measured pre- and post-race to evaluate the biochemical changes in articular cartilage after the subjects run a half-marathon.ResultsA significant increase in the mean ± SD T2 relaxation time was seen in the outer region of the medial tibial plateau (50.1 ± 2.4 versus 54.7 ± 2.6) and there was a significant decrease in T2 relaxation time in the lateral femoral condyle central region (50.2 ± 4.5 versus 45.4 ± 2.9). There were no significant changes in the patella, medial femoral condyle and lateral tibia articular surfaces.ConclusionAn increase in T2 relaxation time occurs in the medial tibial plateau of novice half-marathon runners. This limited region of increased T2 values, when compared with complete medial compartment involvement seen in studies of marathon runners, may represent an association between distance run and changes seen in articular cartilage T2 values.

A simple method for determining the coagulation threshold temperature of transparent tissue-mimicking thermal therapy gel phantoms: Validated by magnetic resonance imaging thermometry.

Tissue-mimicking thermal therapy phantoms that coagulate at specific temperatures are valuable tools for developing and evaluating treatment strategies related to thermal therapy. Here, the authors propose a simple and efficient method for determining the coagulation threshold temperature of transparent thermal therapy gel phantoms. The authors used a previously published gel phantom recipe with 2% (w/v) of bovine serum albumin as the temperature-sensitive protein. Using the programmable heating settings of a polymerase chain reaction (PCR) machine, the authors heated 50 μl gel samples to various temperatures for 3 min and then imaged them using the BioRad Gel Doc system to determine the coagulation temperature using an opacity quantification method. The estimated coagulation temperatures were then validated for gel phantoms prepared with different pH levels using high-intensity focused ultrasound (HIFU) heating and magnetic resonance imaging (MRI) thermometry methods on a clinical MR-HIFU system. The PCR heating method produced consistent and reproducible coagulation of gel samples in precise correlation with the set incubation temperatures. The resulting coagulation threshold temperatures for gel phantoms of varying pH levels were found to be 44.1 ± 0.1, 53.4 ± 0.9, and 60.3 ± 0.9 °C for pH levels of 4.25, 4.50, and 4.75, respectively. This corresponded well with the coagulation threshold temperatures determined by MR-thermometry, with coagulation defined as a 95% decrease in T2 relaxation time, which were estimated at 53.6 ± 1.9 and 62.9 ± 2.4 °C for a pH of 4.50 and 4.75, respectively. The opacity quantification method provides a fast and reproducible estimate of the coagulation threshold temperature of transparent temperature-sensitive gel phantoms. The temperatures determined using this method were well within the range of temperatures estimated using MR-thermometry. Due to the specific heating capabilities of the PCR machine, and the robust determination of coagulation threshold temperatures based on the statistically significant increase in the opacity of gel samples, coagulation temperatures can be determined more precisely and with less variability compared to MRI-based methods.

Quantitative Analysis of Disc Degeneration Using Axial T2 Mapping in a Percutaneous Annular Puncture Model in Rabbits.

ObjectiveTo evaluate T2 relaxation time change using axial T2 mapping in a rabbit degenerated disc model and determine the most correlated variable with histologic score among T2 relaxation time, disc height index, and Pfirrmann grade.Materials and MethodsDegenerated disc model was made in 4 lumbar discs of 11 rabbits (n = 44) by percutaneous annular puncture with various severities of an injury. Lumbar spine lateral radiograph, MR T2 sagittal scan and MR axial T2 mapping were obtained at baseline and 2 weeks and 4 weeks after the injury in 7 rabbits and at baseline and 2 weeks, 4 weeks, and 6 weeks after the injury in 4 rabbits. Generalized estimating equations were used for a longitudinal analysis of changes in T2 relaxation time in degenerated disc model. T2 relaxation time, disc height index and Pfirrmann grade were correlated with the histologic scoring of disc degeneration using Spearman's rho test.ResultsThere was a significant difference in T2 relaxation time between uninjured and injured discs after annular puncture. Progressive decrease in T2 relaxation time was observed in injured discs throughout the study period. Lower T2 relaxation time was observed in the more severely injured discs. T2 relaxation time showed the strongest inverse correlation with the histologic score among the variables investigated (r = -0.811, p < 0.001).ConclusionT2 relaxation time measured with axial T2 mapping in degenerated discs is a potential method to assess disc degeneration.

Characterisation of the antidepressant properties of nitric oxide synthase inhibitors in the olfactory bulbectomised rat model of depression

Nitric oxide synthase (NOS) inhibitors possess antidepressant-like properties in preclinical tests and in the current investigation the brain penetrant NOS inhibitor Nω-nitro-l-arginine (l-NA) and the preferential inhibitor of neuronal NOS (nNOS) 1-(2-trifluoromethylphenyl) imidazole (TRIM) were assessed in the olfactory bulbectomised (OB) rat, a well-established animal model of depression. Magnetic resonance imaging (MRI) was employed to assess regional brain volumes, blood perfusion and T1 and T2 relaxometry times both with and without drug treatment. l-NA (10mg/kg, once daily p.o. for 10 days) attenuated OB-related hyperactivity in the “open field” test in a comparable fashion to the tricyclic antidepressant imipramine (20mg/kg, once daily p.o. for 14 days) indicative of an antidepressant-like response in the model. Treatment with TRIM (50mg/kg, once daily s.c.) attenuated OB-related hyperactivity following 7 days of treatment when compared to vehicle treated controls. OB is associated with enlarged ventricular volume, increased periventicular perfusion and a decrease in T2 relaxation times in cortical and hippocampal regions, with enhanced perfusion and reduced T2 times attenuated by l-NA treatment. l-NA treatment was also associated with an increase in T1 relaxation times in limbic and cortical regions and found to reduce resting state hippocampal blood perfusion in OB animals. Behavioural observations are consistent with an antidepressant action of NOS inhibitors where associated changes in perfusion and T2 relaxation times may be related to the antidepressant action of l-NA in the model.

Age and Sex Dependency of Cartilage T2 Relaxation Time Mapping in MRI of Children and Adolescents

T2 relaxation times on MRI are sensitive to the configuration of cartilage collagen and continually increase during aging in adults. In children, T2 relaxation times increase as a result of cartilage microstructure changes in early inflammatory arthritis. The purpose of this study was to determine age- and sex-related differences in T2 mapping of the patellar cartilage in children and adolescents during normal skeletal maturation. Ninety-seven subjects (age range, 5-22 years; 51 females and 46 males; mean age, 14.3 and 13.7 years, respectively) without patellofemoral instability or inflammatory arthritis were included. All subjects underwent 1.5-T knee MRI with T2 mapping. The mean T2 relaxation time and thickness of the patellar cartilage were documented for each MRI examination. Skeletal maturation was determined by physeal patency (open; or closed or closing) on MRI. The associations between T2 relaxation times, cartilage thickness, sex, age, and physeal patency were assessed using Wilcoxon rank sum test and least-squares means regression models. T2 relaxation times and thickness of the patellar cartilage significantly decreased (p<0.0001) with increasing chronologic age. T2 relaxation times and cartilage thickness in the open physis group were found to be greater than in the closed or closing physis group (p<0.0001). T2 relaxation times and cartilage thickness were greater in males than in females (p<0.05). In contrast to senescent changes in adults, skeletal maturation in children results in a sequential decrease in T2 relaxation times that are age- and sex-dependent. Similar to cartilage in adults, cartilage in children gets progressively thinner during skeletal maturation.

Quantitative T2 mapping of recurrent glioblastoma under bevacizumab improves monitoring for non-enhancing tumor progression and predicts overall survival

Anti-angiogenic treatment in recurrent glioblastoma patients suppresses contrast enhancement and reduces vasogenic edema while non-enhancing tumor progression is common. Thus, the importance of T2-weighted imaging is increasing. We therefore quantified T2 relaxation times, which are the basis for the image contrast on T2-weighted images. Conventional and quantitative MRI procedures were performed on 18 patients with recurrent glioblastoma before treatment with bevacizumab and every 8 weeks thereafter until further tumor progression. We segmented the tumor on conventional MRI into 3 subvolumes: enhancing tumor, non-enhancing tumor, and edema. Using coregistered quantitative maps, we followed changes in T2 relaxation time in each subvolume. Moreover, we generated differential T2 maps by a voxelwise subtraction using the first T2 map under bevacizumab as reference. Visually segmented areas of tumor and edema did not differ in T2 relaxation times. Non-enhancing tumor volume did not decrease after commencement of bevacizumab treatment but strikingly increased at progression. Differential T2 maps clearly showed non-enhancing tumor progression in previously normal brain. T2 relaxation times decreased under bevacizumab without re-increasing at tumor progression. A decrease of <26 ms in the enhancing tumor following exposure to bevacizumab was associated with longer overall survival. Combining quantitative MRI and tumor segmentation improves monitoring of glioblastoma patients under bevacizumab. The degree of change in T2 relaxation time under bevacizumab may be an early response parameter predictive of overall survival. The sustained decrease in T2 relaxation times toward values of healthy tissue masks progressive tumor on conventional T2-weighted images. Therefore, quantitative T2 relaxation times may detect non-enhancing progression better than conventional T2-weighted imaging.

Glial Activation in AβPP/PS1 Mice is Associated with Infiltration of IFNγ-Producing Cells

Whereas the classical histological hallmarks of Alzheimer's disease (AD) are deposition of amyloid-containing plaques and development of neurofibrillary tangles, there is also clear evidence of inflammatory changes accompanied by the presence of activated microglia and astrocytosis. However, at this time, it remains uncertain whether inflammatory changes contribute to pathogenesis of the disease or if they are secondary to deposition of amyloid-β or other pathological changes. A greater understanding of the sequence of events would clearly improve development of strategies to delay progression of the disease. There is a realistic expectation that advances in imaging technology may provide the key to uncovering this sequence. In this study, we employed non-invasive imaging techniques to examine changes in tissue state in hippocampus and cortex of transgenic mice which overexpress amyloid-β protein precursor and presenilin 1 and show that the observed increase in T1 relaxation time was associated with astrogliosis while the decrease in T2 relaxation time was associated with microglial activation. We explored the possibility that interferon-γ might trigger glial activation and demonstrate a genotype-related infiltration of macrophages and natural killer cells, which release interferon-γ. The evidence suggests that IFNγ triggers glial activation and expression of proinflammatory cytokines, and these changes, in turn, contribute to the decrease in long-term potentiation.

An interplay between electrostatic and polar interactions in peptide hydrogels.

Inherent chemical programmability available in peptide-based hydrogels has allowed diversity in the development of these materials for use in biomedical applications. Within the 20 natural amino acids, a range of chemical moieties are present. Here we used a mixing-induced self-assembly of two oppositely charged peptide modules to form a peptide-based hydrogel. To investigate electrostatic and polar interactions in the hydrogel, we replace amino acids from the negatively charged acidic glutamic acid (E) to the uncharged polar glutamine (Q) on a negatively charged peptide module, while leaving the positively charged module unchanged. Using dynamic rheology, the mechanical properties of each hydrogel were investigated. It was found that the number, but not the location, of electrostatic interactions (E residues) dictate the elastic modulus (G') of the hydrogel, compared to polar interactions (Q residues). Increased electrostatic interactions also promote faster peptide assembly into the hydrogel matrix, and result in the decrease of T2 relaxation times of H2 O and trifluoroacetic acid. Small-angle X-ray scattering (SAXS) showed that changing from electrostatic to polar interactions affects the ability to form fibrous networks: from the formation of elongated fibers to no fiber assembly. This study reveals the systematic effects that the incorporation of electrostatic and polar interactions have when programmed into peptide-based hydrogel systems. These effects could be used to design peptide-based biomaterials with predetermined properties.

Ferritin Overexpression for Noninvasive Magnetic Resonance Imaging–Based Tracking of Stem Cells Transplanted into the Heart

An unmet need in cardiac cell therapy is a noninvasive imaging technique capable of tracking changes in graft size over time and monitoring cell dynamics such as replication and death, factors to which commonly used superparamagnetic nanoparticles are insensitive. Our goal was to explore if overexpression of ferritin, a nontoxic iron-binding protein, can be used for noninvasive magnetic resonance imaging (MRI) of cells transplanted into the infarcted heart. Mouse skeletal myoblasts (C2C12 cells) were engineered to overexpress ferritin. Ferritin overexpression did not interfere with cell viability, proliferation, or differentiation into multinucleated myotubes. Ferritin overexpression caused a 25% decrease in T2 relaxation time in vitro compared to wild-type cells. Transgenic grafts were detected in vivo 3 weeks after transplantation into infarcted hearts of syngeneic mice as areas of hypointensity caused by iron accumulation in overexpressed ferritin complexes. Graft size evaluation by MRI correlated tighly with histologic measurements (R2 = .8). Our studies demonstrated the feasibility of ferritin overexpression in mouse skeletal myoblasts and the successful detection of transgenic cells by MRI in vitro and in vivo after transplantation into the infarcted mouse heart. These experiments lay the groundwork for using the MRI gene reporter ferritin to track stem cells transplanted to the heart.

Renal Inflammation: Targeted Iron Oxide Nanoparticles for Molecular MR Imaging in Mice

To determine the feasibility of T2-weighted magnetic resonance (MR) imaging in the noninvasive quantification of renal inflammation by using superparamagnetic iron oxide (SPIO) nanoparticles targeted to tissue-bound C3 activation fragments in a mouse model of lupus nephritis. All animal procedures were approved by the University of Colorado-Denver animal care and use committee. SPIO nanoparticles were encapsulated by using amine-functionalized phospholipids. A recombinant protein containing the C3d-binding region of complement receptor type 2 (CR2) was then conjugated to the surface of the SPIO nanoparticle. Five MRL/lpr mice (a model of lupus nephritis) and six C57BL/6 wild-type mice were assessed with T2-weighted MR imaging at baseline and after SPIO injection. The same five MRL/lpr mice and three C57BL/6 mice also underwent MR imaging after injection of CR2-targeted SPIO. A series of T2-weighted pulses with 16 echo times was used to enable precise T2 mapping and calculation of T2 relaxation times in the cortex and outer and inner medulla of the kidneys, as well as in the spleen, muscle, and fat. The effects of treatment and animal genotype on T2 relaxation times were analyzed with repeated-measures analysis of variance. At baseline, the T2-weighted signal intensity in the kidneys of MRL/lpr mice was higher than that in the kidneys of wild-type mice. Injection of untargeted SPIO did not alter the T2-weighted signal in the kidneys in either strain of mice. Injection of CR2-targeted SPIO in MRL/lpr mice, however, caused a significant accumulation of targeted iron oxide with a subsequent decrease in T2 relaxation times in the cortex and outer and inner medulla of the kidneys. No changes in T2 relaxation time were observed in the wild-type mice after injection of targeted SPIO. Injection of CR2-conjugated SPIO caused a significant reduction in T2-weighted MR imaging signal and T2 relaxation time in nephritic kidneys.

MR Reporter Gene Imaging of Endostatin Expression and Therapy

The aim of this study is to monitor endostatin gene expression and therapy using transferrin receptor (TfR) as reporter gene and transferrin conjugate of ultrasmall supramagnetic iron oxide nanoparticle (Tf-USPIO) as magnetic resonance (MR) reporter probe. A retroviral plasmid (pLP-LNCX) encoding mouse endostatin and TfR was constructed, and packaged with a titer of 4 × 10(7)colony-forming units per millimeter. MDA-MB-231 breast tumors were established in BALB/c mice by subcutaneous injection of 2 × 10(6) MDA-MB-231 cells. Mice were intratumorally injected with recombinant retrovirus and imaged with MR using Tf-USPIO. Western blot, Prussian blue, and immunohistochemical staining were performed to validate the magnetic resonance imaging results. The antitumor effect of retro-endostatin (ES)-TfR was also evaluated by intratumoral injection of the viral vector. The expression of both endostatin and TfR genes in MDA-MB-231 cells after retroviral transfection was confirmed by Western blot and flow cytometry. Tf-USPIO conjugate binds specifically to cells stably transfected with retro-ES-TfR. After intravenous injection of the Tf-USPIO conjugate, there was a more pronounced decrease in T2 relaxation time in tumors treated with retro-ES-TfR than in tumors treated with empty retrovirus retro-LNCX. The expression of ES gene significantly delayed the growth of MDA-MB-231 tumor and reduction of microvessel density and VEGF level as compared to those without viral transfection or transfected with empty retro-LNCX vector. Endostatin therapeutic gene expression was visualized successfully using TfR reporter gene and Tf-USPIO MR reporter probe, which indicates that MR reporter gene imaging may be valuable in gene therapy to evaluate therapeutic gene expression and treatment efficacy.

Specific Targeting of Tumor Angiogenesis by RGD-Conjugated Ultrasmall Superparamagnetic Iron Oxide Particles Using a Clinical 1.5-T Magnetic Resonance Scanner

Angiogenesis is essential for the development of malignant tumors and provides important targets for tumor diagnosis and therapy. To noninvasively assess the angiogenic profile of tumors, novel alpha(v)beta(3) integrin-targeted ultrasmall superparamagnetic iron oxide particles (USPIOs) were designed and their specific uptake by endothelial cells was evaluated in vitro and in vivo. USPIOs were coated with 3-aminopropyltrimethoxysilane (APTMS) and conjugated with Arg-Gly-Asp (RGD) peptides. Accumulation in human umbilical vein endothelial cells (HUVECs) was evaluated using Prussian blue staining, transmission electron microscopy, magnetic resonance (MR) imaging, and inductively coupled plasma mass spectrometry. Uptake of RGD-USPIO by HUVECs was significantly increased when compared with unlabeled USPIO and could be competitively inhibited by addition of unbound RGD. The ability of the RGD-USPIO to noninvasively distinguish tumors with high (HaCaT-ras-A-5RT3) and lower (A431) area fractions of alpha(v)beta(3) integrin-positive vessels was evaluated using a 1.5-T MR scanner. Indeed, after RGD-USPIO injection, there was a more pronounced decrease in T(2) relaxation times in HaCaT-ras-A-5RT3 tumors than in A431 tumors. Furthermore, T(2)*-weighted images clearly identified the heterogeneous arrangement of vessels with alpha(v)beta(3) integrins in HaCaT-ras-A-5RT3 tumors by an irregular signal intensity decrease. In contrast, in A431 tumors with predominantly small and uniformly distributed vessels, the signal intensity decreased more homogeneously. In summary, RGD-coupled, APTMS-coated USPIOs efficiently label alpha(v)beta(3) integrins expressed on endothelial cells. Furthermore, these molecular MR imaging probes are capable of distinguishing tumors differing in the degree of alpha(v)beta(3) integrin expression and in their angiogenesis profile even when using a clinical 1.5-T MR scanner.

Influence of Aging and Salting on Protein Secondary Structures and Water Distribution in Uncooked and Cooked Pork. A Combined FT-IR Microspectroscopy and 1H NMR Relaxometry Study

Fourier transform infrared (FT-IR) microspectroscopy and low-field (LF) proton NMR transverse relaxation measurements were used to study the changes in protein secondary structure and water distribution as a consequence of aging (1 day and 14 days) followed by salting (3%, 6%, and 9% NaCl) and cooking (65 degrees C). An enhanced water uptake and increased proton NMR relaxation times after salting were observed in aged meat (14 days) compared with nonaged meat (1 day). FT-IR bands revealed that salting induced an increase in native beta-sheet structure while aging triggered an increase in native alpha-helical structure before cooking, which could explain the effects of aging and salting on water distribution and water uptake. Moreover, the decrease in T2 relaxation times and loss of water upon cooking were attributed to an increase in aggregated beta-sheet structures and a simultaneous decrease in native protein structures. Finally, aging increased the cooking loss and subsequently decreased the final yield, which corresponded to a further decrease in T2 relaxation times in aged meat upon cooking. However, salting weakened the effect of aging on the final yield, which is consistent with the increased T2 relaxation times upon salting for aged meat after cooking and the weaker effect of aging on protein secondary structural changes for samples treated with high salt concentration. The present study reveals that changes in water distribution during aging, salting, and cooking are not only due to the accepted causal connection, i.e., proteolytic degradation of myofibrillar structures, change in electrostatic repulsion, and dissolution and denaturation of proteins, but also dynamic changes in specific protein secondary structures.