Lauriane Jugé

Changes in Rat Brain Tissue Microstructure and Stiffness during the Development of Experimental Obstructive Hydrocephalus.

Understanding neural injury in hydrocephalus and how the brain changes during the course of the disease in-vivo remain unclear. This study describes brain deformation, microstructural and mechanical properties changes during obstructive hydrocephalus development in a rat model using multimodal magnetic resonance (MR) imaging. Hydrocephalus was induced in eight Sprague-Dawley rats (4 weeks old) by injecting a kaolin suspension into the cisterna magna. Six sham-injected rats were used as controls. MR imaging (9.4T, Bruker) was performed 1 day before, and at 3, 7 and 16 days post injection. T2-weighted MR images were collected to quantify brain deformation. MR elastography was used to measure brain stiffness, and diffusion tensor imaging (DTI) was conducted to observe brain tissue microstructure. Results showed that the enlargement of the ventricular system was associated with a decrease in the cortical gray matter thickness and caudate-putamen cross-sectional area (P < 0.001, for both), an alteration of the corpus callosum and periventricular white matter microstructure (CC+PVWM) and rearrangement of the cortical gray matter microstructure (P < 0.001, for both), while compression without gross microstructural alteration was evident in the caudate-putamen and ventral internal capsule (P < 0.001, for both). During hydrocephalus development, increased space between the white matter tracts was observed in the CC+PVWM (P < 0.001), while a decrease in space was observed for the ventral internal capsule (P < 0.001). For the cortical gray matter, an increase in extracellular tissue water was significantly associated with a decrease in tissue stiffness (P = 0.001). To conclude, this study characterizes the temporal changes in tissue microstructure, water content and stiffness in different brain regions and their association with ventricular enlargement. In summary, whilst diffusion changes were larger and statistically significant for majority of the brain regions studied, the changes in mechanical properties were modest. Moreover, the effect of ventricular enlargement is not limited to the CC+PVWM and ventral internal capsule, the extent of microstructural changes vary between brain regions, and there is regional and temporal variation in brain tissue stiffness during hydrocephalus development.

Liver Stiffness Values Are Lower in Pediatric Subjects than in Adults and Increase with Age: A Multifrequency MR Elastography Study

Purpose To determine if healthy hepatic mechanical properties differ between pediatric and adult subjects at magnetic resonance (MR) elastography. Materials and Methods Liver shear moduli in 24 healthy pediatric participants (13 children aged 5-14 years [seven boys, six girls] and 11 adolescents aged 15-18 years [six boys, five girls]) and 10 healthy adults (aged 22-36 years [five men, five women]) were obtained with 3-T MR elastography at 28, 56, and 84 Hz. Relationships between shear moduli and age were assessed with Spearman correlations. Differences between age groups were determined with one-way analysis of variance and Tukey multiple comparisons tests. Results Liver stiffness values (means ± standard deviations) were significantly lower in children and adolescents than in adults at 56 Hz (children, 2.2 kPa ± 0.3; adolescents, 2.2 kPa ± 0.2; adults, 2.6 kPa ± 0.3; analysis of variance, P = .009) and 84 Hz (children, 5.6 kPa ± 0.8; adolescents, 6.5 kPa ± 1.2; adults, 7.8 kPa ± 1.2; analysis of variance, P = .0003) but not at 28 Hz (children, 1.2 kPa ± 0.2; adolescents, 1.3 kPa ± 0.3; adults, 1.2 kPa ± 0.2; analysis of variance, P = .40). At 56 and 84 Hz, liver stiffness increased with age (Spearman correlation, r = 0.38 [P = .03] and r = 0.54 [P = .001], respectively). Stiffness varied less with frequency in children and adolescents than in adults (analysis of variance, P = .0009). No significant differences were found in shear moduli at 28, 56, or 84 Hz or frequency dependence between children and adolescents (P = .38, P = .99, P = .14, and P = .30, respectively, according to Tukey tests). Conclusion Liver stiffness values are lower and vary less with frequency in children and adolescents than in adults. Stiffness increases with age during normal development and approaches adult values during adolescence. Comparing pediatric liver stiffness to adult baseline values to detect pediatric liver mechanical abnormalities may not allow detection of mild disease and may lead to underestimation of severity.

Microvasculature alters the dispersion properties of shear waves - a multi-frequency MR elastography study

Magnetic Resonance Elastography (MRE) uses macroscopic shear wave propagation to quantify mechanical properties of soft tissues. Micro‐obstacles are capable of affecting the macroscopic dispersion properties of shear waves. Since disease or therapy can change the mechanical integrity and organization of vascular structures, MRE should be able to sense these changes if blood vessels represent a source for wave scattering. To verify this, MRE was performed to quantify alteration of the shear wave speed cs due to the presence of vascular outgrowths using an aortic ring model. Eighteen fragments of rat aorta included in a Matrigel matrix (n=6 without outgrowths, n=6 with a radial outgrowth extent of ~600µm and n=6 with ~850µm) were imaged using a 7 Tesla MR scanner (Bruker, PharmaScan). High resolution anatomical images were acquired in addition to multi‐frequency MRE (ν = 100, 115, 125, 135 and 150 Hz). Average cs was measured within a ring of ~900µm thickness encompassing the aorta and were normalized to cs0 of the corresponding Matrigel. The frequency dependence was fit to the power law model cs ~νy. After scanning, optical microscopy was performed to visualize outgrowths. Results demonstrated that in presence of vascular outgrowths (1) normalized cs significantly increased for the three highest frequencies (Kruskal‐Wallis test, P = 0.0002 at 125 Hz and P = 0.002 at 135 Hz and P = 0.003 at 150 Hz) but not for the two lowest (Kruskal‐Wallis test, P = 0.63 at 100 Hz and P = 0.87 at 115 Hz), and (2) normalized cs followed a power law behavior not seen in absence of vascular outgrowths (ANOVA test, P < 0.0001). These results showed that vascular outgrowths acted as micro‐obstacles altering the dispersion relationships of propagating shear waves and that MRE could provide valuable information about microvascular changes. Copyright

Development of acute hydrocephalus does not change brain tissue mechanical properties in adult rats, but in juvenile rats

Introduction Regional changes in brain stiffness were previously demonstrated in an experimental obstructive hydrocephalus juvenile rat model. The open cranial sutures in the juvenile rats have influenced brain compression and mechanical properties during hydrocephalus development and the extent by which closed cranial sutures in adult hydrocephalic rat models affect brain stiffness in-vivo remains unclear. The aims of this study were to determine changes in brain tissue mechanical properties and brain structure size during hydrocephalus development in adult rat with fixed cranial volume and how these changes were related to brain tissue deformation. Methods Hydrocephalus was induced in 9 female ten weeks old Sprague-Dawley rats by injecting 60 μL of a kaolin suspension (25%) into the cisterna magna under anaesthesia. 6 sham-injected age-matched female SD rats were used as controls. MR imaging (9.4T, Bruker) was performed 1 day before and then at 3 days post injection. T2-weighted anatomical MR images were collected to quantify ventricle and brain tissue cross-sectional areas. MR elastography (800 Hz) was used to measure the brain stiffness (G*, shear modulus). Results Brain tissue in the adult hydrocephalic rats was more compressed than the juvenile hydrocephalic rats because the skulls of the adult hydrocephalic rats were unable to expand like the juvenile rats. In the adult hydrocephalic rats, the cortical gray matter thickness and the caudate-putamen cross-sectional area decreased (Spearman, P < 0.001 for both) but there were no significant changes in cranial cross-sectional area (Spearman, P = 0.35), cortical gray matter stiffness (Spearman, P = 0.24) and caudate-putamen (Spearman, P = 0.11) stiffness. No significant changes in the size of brain structures were observed in the controls. Conclusions This study showed that although brain tissue in the adult hydrocephalic rats was severely compressed, their brain tissue stiffness did not change significantly. These results are in contrast with our previous findings in juvenile hydrocephalic rats which had significantly less brain compression (as the brain circumference was able to stretch with the cranium due to the open skull sutures) and had a significant increase in caudate putamen stiffness. These results suggest that change in brain mechanical properties in hydrocephalus is complex and is not solely dependent on brain tissue deformation. Further studies on the interactions between brain tissue stiffness, deformation, tissue oedema and neural damage are necessary before MRE can be used as a tool to track changes in brain biomechanics in hydrocephalus.

Evaluation of Nonradiative Clinical Imaging Techniques for the Longitudinal Assessment of Tumour Growth in Murine CT26 Colon Carcinoma

Background and Objectives. To determine the most appropriate technique for tumour followup in experimental therapeutics, we compared ultrasound (US) and magnetic resonance imaging (MRI) to characterize ectopic and orthotopic colon carcinoma models. Methods. CT26 tumours were implanted subcutaneously (s.c.) in Balb/c mice for the ectopic model or into the caecum for the orthotopic model. Tumours were evaluated by histology, spectrofluorescence, MRI, and US. Results. Histology of CT26 tumour showed homogeneously dispersed cancer cells and blood vessels. The visualization of the vascular network using labelled albumin showed that CT26 tumours were highly vascularized and disorganized. MRI allowed high-resolution and accurate 3D tumour measurements and provided additional anatomical and functional information. Noninvasive US imaging allowed good delineation of tumours despite an hypoechogenic signal. Monitoring of tumour growth with US could be accomplished as early as 5 days after implantation with a shorter acquisition time (<5 min) compared to MRI. Conclusion. MRI and US afforded excellent noninvasive imaging techniques to accurately follow tumour growth of ectopic and orthotopic CT26 tumours. These two techniques can be appropriately used for tumour treatment followup, with a preference for US imaging, due to its short acquisition time and simplicity of use.

Effect of endoscopic third ventriculostomy on cerebrospinal fluid pressure in the cerebral ventricles

We aimed to show how endoscopic third ventriculostomy (ETV) treatment may affect cerebrospinal fluid (CSF) flow dynamics in hydrocephalus, with and without aqueductal stenosis. Hydrocephalus is a neurological disorder which is characterized by enlarged brain ventricles. The periodic motion of CSF flow as a function of the cardiac cycle was prescribed as the inlet boundary condition at the foramen of Monro, and ETV was modeled as a 5mm diameter hole in the anterior wall of the third ventricle. The results show that ETV reduces the pressure in the ventricles by nine-fold in the model with aqueductal stenosis, and three-fold in the model without aqueductal stenosis. More importantly, ETV changes the temporal characteristics of the CSF pressure waveform in the model without aqueductal stenosis, such that there is higher pressure in the ventricle during diastole. This study suggests that changes in the temporal characteristics of the CSF pressure waveform in the ventricles may be the reason why ETV treatment is not effective for hydrocephalus without aqueductal stenosis.

Lipidic spherulites as magnetic resonance imaging contrast agents

Magnetic resonance imaging is an excellent technique to achieve anatomical details and highly resolved images. The search for efficient contrast agents to increase the signal to background ratio led us to evaluate paramagnetic spherulites as potential Magnetic Resonance Imaging (MRI) contrast agents. Spherulites are supramolecular assemblies, made of lipidic concentric multilayers, able to encapsulate with high efficiency soluble macromolecules. Despite their highly interesting structure, spherulites have never been proposed as imaging agents. We proposed here three approaches to render spherulites paramagnetic: encapsulating a soluble contrastophore, inserting a lipidic contrastophore derivative or grafting a soluble contrastophore at the surface of the spherulites. Following similar strategies, liposomes were prepared for comparison. The conservation of the spherulite structure, throughout these three strategies, was shown by cryoelectron microscopy and small angle light scattering. The effect of the paramagnetic spherulites was studied by magnetic resonance imaging at different magnetic fields. The results showed that insertion of a contrastophore lipidic derivative into spherulite bilayers and grafting a contrastophore at the surface of the spherulites were the two strategies which led to the highest MRI contrast improvement.

Characterising skeletal muscle under large strain using eccentric and Fourier Transform-rheology

Characterising the passive anisotropic properties of soft tissues has been largely limited to the linear viscoelastic regime and shear loading is rarely done in the large deformation regime, despite the physiological significance of such properties. This paper demonstrates the use of eccentric rheology, which allows the anisotropy of skeletal muscle to be investigated. The large amplitude oscillatory strain properties of skeletal muscle were also investigated using Fourier Transform-rheology. Histology was used to qualitatively assess the microstructure changes induced by large strain. Results showed that skeletal muscle was strongly anisotropic in the linear regime. The storage and loss moduli were found to be significantly different (p<0.05) between the three fibre alignment groups; for the group tested with fibres perpendicular to plane of shear was 12.3±1.3 kPa and 3.0±0.35 kPa, parallel to shear direction was 10.6±1.2 kPa and 2.4±0.23 kPa, and perpendicular to shear direction was 5.5±0.90 kPa and 1.3±0.21 kPa. The appearance and growth of higher order harmonics at large strain was different in the three testing directions indicating that the anisotropy of muscle affects skeletal muscle behaviour in the nonlinear regime. Histological analysis showed an increasing destruction of extracellular matrix and the rearrangement of fibres with increasing strain indicating mechanical damage at strains of larger than 10%. These microstructural changes could contribute to the complex nonlinear behaviour in skeletal muscle. This paper demonstrates a method of characterising the anisotropic properties in skeletal muscle under large strain whilst giving meaningful information on the physical response of tissue at various strains.

Covertly active and progressing neurochemical abnormalities in suppressed HIV infection

Objective To assess whether HIV-related brain injury is progressive in persons with suppressed HIV infection. Methods Seventy-three HIV+ virally suppressed men and 35 HIV− men, screened for psychiatric and alcohol/drug use disorders, underwent neuropsychological evaluation and proton magnetic resonance spectroscopy (1H-MRS) at baseline and after and 23 ± 5 months. 1H-MRS included brain regions known to be vulnerable to HIV and aging: frontal white matter (FWM), posterior cingulate cortex (PCC), and caudate area (CA). Major brain metabolites such as creatine (Cr: marker of cellular energy), N-acetyl aspartate (NAA: marker of neuronal integrity), choline (marker of cellular membrane turnover), glutamate/glutamine (excitatory/inhibitory neurotransmitter), and myo-Inositol (mI: marker of neuroinflammation) were calculated with reference to water signal. Neurocognitive decline was corrected for practice effect and baseline HIV-associated neurocognitive disorder (HAND) status. Results Across the study period, 44% had intact cognition, 42% stable HAND (including the single case that improved), 10% progressing HAND, and 4% incident HAND. When analyzing the neurochemical data per neurocognitive trajectories, we found decreasing PCC Cr in all subgroups compared with controls (p < 0.002). In addition, relative to the HIV− group, stable HAND showed decreasing FWM Cr, incident HAND showed steep FWM Cr reduction, whereas progressing HAND had a sharply decreasing PCC NAA and reduced but stable CA NAA. When analyzing the neurochemical data at the group level (HIV+ vs HIV− groups), we found stable abnormal metabolite concentrations over the study period: decreased FWM and PCC Cr (both p < 0.001), decreased PCC NAA and CA NAA (both p < 0.05) and PCC mI increase (p < 0.05). HIV duration and historical HAND had modest effects on metabolite changes. Conclusions Our study reveals covertly active or progressing HIV-related brain injury in the majority of this virally suppressed cohort, reflecting ongoing neuropathogenic processes that are only partially worsened by historical HAND and HIV duration. Longer-term studies will be important for determining the prognosis of these slowly evolving neurochemical abnormalities.

Measurement of large strain properties in calf muscles in vivo using magnetic resonance elastography and spatial modulation of magnetization

It is important to measure the large deformation properties of skeletal muscle in vivo in order to understand and model movement and the force‐producing capabilities of muscle. As muscle properties are non‐linear, an understanding of how the deformation state affects the measured shear moduli is also useful for clinical applications of magnetic resonance elastography (MRE) to muscle disorders. MRE has so far only been used to measure the linear viscoelastic (small strain) properties of muscles. This study aims to measure the shear moduli of human calf muscles under varying degrees of strain using MRE. Nine healthy adults (four males; age range, 25–38 years) were recruited, and the storage modulus G′ was measured at three ankle angle positions: P0 (neutral), P15 (15° plantarflexed) and P30 (30° plantarflexed). Spatial modulation of magnetization (SPAMM) was used to measure the strain in the calf associated with the ankle rotations between P0 to P15 and P0 to P30. SPAMM results showed that, with plantarflexion, there was a shortening of the medial gastrocnemius and soleus muscles, which resulted in an expansion of both muscles in the transverse direction. Strains for each ankle rotation were in the range 3–9% (in compression). MRE results showed that this shortening during plantarflexion resulted in a mean decrease in G′ in the medial gastrocnemius (p = 0.013, linear mixed model), but not in the soleus (p = 0.47). This study showed that MRE is a viable technique for the measurement of large strain deformation properties in vivo in soft tissues by inducing physiological strain within the muscle during imaging.