Influence of pia-arachnoid complex on the indentation response of porcine brain at different length scales.

Brain tissues are surrounded by two tightly adhering thin membranes known as the pia-arachnoid complex (PAC), which is pivotal in regulating brain mechanical response upon mechanical impact. Despite the crucial role of PAC as a structural damper protecting the brain, its mechanical contribution has received minimal attention. In this work, the mechanical contribution of PAC on brain tissues against mechanical loading is characterized by using a custom-built indentation apparatus. The indentation responses of the isolated and PAC-overlaid brains are quantitatively compared at different length scales and strain rates. Results show that PAC substantially affects the indentation response of brain tissues at micro- and macro-scales and provides better protection against mechanical impact at a relatively small (µm) length scale. The modulus of the PAC-overlaid brain shows a threefold stiffening at the microscale compared with that of the isolated brain (with instantaneous shear modulus distribution means of 0.85 ± 0.14 kPa versus 2.64 ± 0.43 kPa at the strain rate of 0.64 s-1 and 1.40 ± 0.31 kPa versus 4.02 ± 0.51 at 1.27 s-1). These findings indicate that PAC seriously affects the mechanical response of brain tissues, especially at the microscale, and may have important implications for the studies of brain injury.

Mechanical Properties of the Cranial Meninges: A Systematic Review.

The meninges are membranous tissues that are pivotal in maintaining homeostasis of the central nervous system. Despite the importance of the cranial meninges in nervous system physiology and in head injury mechanics, our knowledge of the tissues' mechanical behavior and structural composition is limited. This systematic review analyzes the existing literature on the mechanical properties of the meningeal tissues. Publications were identified from a search of Scopus, Academic Search Complete, and Web of Science and screened for eligibility according to Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. The review details the wide range of testing techniques employed to date and the significant variability in the observed experimental findings. Our findings identify many gaps in the current literature that can serve as a guide for future work for meningeal mechanics investigators. The review identifies no peer-reviewed mechanical data on the falx and tentorium tissues, both of which have been identified as key structures in influencing brain injury mechanics. A dearth of mechanical data for the pia-arachnoid complex also was identified (no experimental mechanics studies on the human pia-arachnoid complex were identified), which is desirable for biofidelic modeling of human head injuries. Finally, this review provides recommendations on how experiments can be conducted to allow for standardization of test methodologies, enabling simplified comparisons and conclusions on meningeal mechanics.

Ex-vivo quantification of ovine pia arachnoid complex biomechanical properties under uniaxial tension.

BACKGROUND: The pia arachnoid complex (PAC) is a cerebrospinal fluid-filled tissue conglomerate that surrounds the brain and spinal cord. Pia mater adheres directly to the surface of the brain while the arachnoid mater adheres to the deep surface of the dura mater. Collagen fibers, known as subarachnoid trabeculae (SAT) fibers, and microvascular structure lie intermediately to the pia and arachnoid meninges. Due to its structural role, alterations to the biomechanical properties of the PAC may change surface stress loading in traumatic brain injury (TBI) caused by sub-concussive hits. The aim of this study was to quantify the mechanical and morphological properties of ovine PAC. METHODS: Ovine brain samples (n = 10) were removed from the skull and tissue was harvested within 30 min post-mortem. To access the PAC, ovine skulls were split medially from the occipital region down the nasal bone on the superior and inferior aspects of the skull. A template was used to remove arachnoid samples from the left and right sides of the frontal and occipital regions of the brain. 10 ex-vivo samples were tested with uniaxial tension at 2 mm s-1, average strain rate of 0.59 s-1, until failure at < 5 h post extraction. The force and displacement data were acquired at 100 Hz. PAC tissue collagen fiber microstructure was characterized using second-harmonic generation (SHG) imaging on a subset of n = 4 stained tissue samples. To differentiate transverse blood vessels from SAT by visualization of cell nuclei and endothelial cells, samples were stained with DAPI and anti-von Willebrand Factor, respectively. The Mooney-Rivlin model for average stress-strain curve fit was used to model PAC material properties. RESULTS: The elastic modulus, ultimate stress, and ultimate strain were found to be 7.7 ± 3.0, 2.7 ± 0.76 MPa, and 0.60 ± 0.13, respectively. No statistical significance was found across brain dissection locations in terms of biomechanical properties. SHG images were post-processed to obtain average SAT fiber intersection density, concentration, porosity, tortuosity, segment length, orientation, radial counts, and diameter as 0.23, 26.14, 73.86%, 1.07 ± 0.28, 17.33 ± 15.25 µm, 84.66 ± 49.18°, 8.15%, 3.46 ± 1.62 µm, respectively. CONCLUSION: For the sizes, strain, and strain rates tested, our results suggest that ovine PAC mechanical behavior is isotropic, and that the Mooney-Rivlin model is an appropriate curve-fitting constitutive equation for obtaining material parameters of PAC tissues.

Hyper-viscoelastic mechanical behavior of cranial pia mater in tension.

BACKGROUND: Cranial pia mater, the innermost layer of the meninges, protects the central nervous system by tightly wrapping the brain and damping the external impact force to the brain. Accurate experimental data of the mechanical property of the cranial pia mater can enhance the theoretical prediction of traumatic brain injury or the scientific surgery design for brain disease. The aim of this study is to characterize the mechanical behavior of the cranial pia mater. METHODS: In vitro tensile and stress-relaxation experiments of ovine cranial pia mater specimens were conducted at eight strain rates to characterize the rate-dependent viscoelastic property. The tensile and stress-relaxation experimental data were fitted by an Ogden hyper-viscoelastic model with a strain rate function to describe the mechanical behavior of the cranial pia mater. FINDINGS: The elastic modulus and the ultimate stress are significantly increased from 5.545 MPa and 0.535 MPa at 0.00167 s-1 to 18.345 MPa and 2.547 MPa at 0.83 s-1 (p < .0001), respectively. The initial stress and the long-term stress (300 s) are also increased significantly with the increasing strain rates (p < .0001). A good fit of the experimental data with the Ogden hyper-viscoelastic model incorporated with a strain rate function was achieved (R2 > 0.93). INTERPRETATION: The cranial pia mater exhibits as a rate-dependent hyper-viscoelastic material in the tensile and stress-relaxation experiments. Compared with the brain, the stiffer nature of the cranial pia mater indicates its essential role in brain protection. The rate-dependent constitutive model provides a proper description of the hyper-viscoelastic characteristics of the cranial pia mater in tension and may provide a basic constitutive relationship for numerical simulations of traumatic brain injury.

Estimates of cortical column orientation improve MEG source inversion.

Determining the anatomical source of brain activity non-invasively measured from EEG or MEG sensors is challenging. In order to simplify the source localization problem, many techniques introduce the assumption that current sources lie on the cortical surface. Another common assumption is that this current flow is orthogonal to the cortical surface, thereby approximating the orientation of cortical columns. However, it is not clear which cortical surface to use to define the current source locations, and normal vectors computed from a single cortical surface may not be the best approximation to the orientation of cortical columns. We compared three different surface location priors and five different approaches for estimating dipole vector orientation, both in simulations and visual and motor evoked MEG responses. We show that models with source locations on the white matter surface and using methods based on establishing correspondences between white matter and pial cortical surfaces dramatically outperform models with source locations on the pial or combined pial/white surfaces and which use methods based on the geometry of a single cortical surface in fitting evoked visual and motor responses. These methods can be easily implemented and adopted in most M/EEG analysis pipelines, with the potential to significantly improve source localization of evoked responses.

Investigation of wave propagation through head layers with focus on understanding blast wave transmission.

Blast-induced traumatic brain injury (bTBI) is a critical health concern. This issue is being addressed in terms of identifying a cause-effect relationship between the mechanical insult in the form of a blast and resulting injury to the brain. Understanding wave propagation through the head is an important aspect in this regard. The objective of this work was to study the blast wave propagation through the layered architecture of the head with an emphasis on understanding the wave transmission mechanism. Toward this end, one-dimensional (1D) finite element head model is built for a simplified surrogate, human, and rat. Motivated from experimental investigations, four different head layer configurations have been considered. These configurations are: (A) Skull-Brain, (B) Skin-Skull-Brain, (C) Skin-Skull-Dura-Arachnoid-CSF-Pia-Brain, (D) Skin-Skull-Dura-Arachnoid-AT-Pia-Brain. The validated head model is subjected to flattop and Friedlander loading implied in the blast, and the resulting response is evaluated in terms of brain pressures. Our results suggest that wave propagation through head parenchyma plays an important role in blast wave transmission. The thickness, material properties of head layers, and rise time of an input pulse govern the temporal evolution of pressure in the brain. The key findings of this work are: (a) Skin and meninges amplify the applied input pressure, whereas air sinus has an attenuation effect. (b) Model is able to describe experimentally recorded peak pressures and rise times in the brain, including variations within the aforementioned experimental head models of TBI. This reinforces that the wave transmission is an important loading pathway to the brain. (c) Equivalent layer theory for modeling meningeal layers as a single layer has been proposed, and it gives reasonable agreement with each meningeal layer modeled explicitly. This modeling approach has a great utility in 3D head models. The potential applications of 1D head model in evaluation of new helmet materials, brain sensor calibration, and brain pressure estimation for a given explosive strength have also been demonstrated. Overall, these results provide important insights into the understanding of mechanics of blast wave transmission in the head.

Micromechanical heterogeneity of the rat pia-arachnoid complex.

To better understand the onset of damage occurring in the brain upon traumatic events, it is essential to analyze how external mechanical loads propagate through the skull and meninges and down to the brain cortex. However, despite their crucial role as structural dampers protecting the brain, the mechanical properties and dynamic behavior of the meningeal layers are still poorly understood. Here, we characterized the local mechanical heterogeneity of rat pia-arachnoid complex (PAC) at the microscale via atomic force microscopy (AFM) indentation experiments to understand how microstructural variations at the tissue level can differentially affect load propagation. By coupling AFM mechanical testing with fresh tissue immunofluorescent staining, we could directly observe the effect of specific anatomical features on the local mechanical properties of tissue. We observed a two-fold stiffening of vascularized tissue when compared to non-vascularized PAC (with instantaneous Young's modulus distribution means of 1.32  ±â€¯ 0.03 kPa and 2.79  ±â€¯ 0.08 kPa, respectively), and statistically significant differences between regions of low- and high-vimentin density, reflecting trabecular density (with means of 0.67  ±â€¯ 0.05 kPa and 1.29  ±â€¯ 0.06 kPa, respectively). No significant differences were observed between cortical and cerebellar PAC. Additionally, by performing force relaxation experiments at the AFM, we identified the characteristic time constant τ1 of PAC tissue to be in the range of 2.7  ±â€¯ 1.2 s to 3.1  ±â€¯ 0.9 s for the different PAC regions analyzed. Taken together, the results presented point at the complex biomechanical nature of the meningeal tissue, and underscore the need to account for its heterogeneity when modeling its behavior into finite element simulations or other computational methods enabling the prediction of load propagation during injury events. STATEMENT OF SIGNIFICANCE: The meningeal layers are pivotal in shielding the brain during injury events, yet the mechanical properties of this complex biological interface are still under investigation. Here, we present the first anatomically-informed micromechanical characterization of mammalian pia-arachnoid complex (PAC). We developed a protocol for the isolation and fresh immunostaining of rat PAC and subjected the tissue to AFM indentation and relaxation experiments, while visualizing the local anatomy via fluorescence microscopy. We found statistically significant variations in regional PAC stiffness according to the degree of vascularization and trabecular cell density, besides identifying the tissue's characteristic relaxation constant. In essence, this study captures the relationship between anatomy and mechanical heterogeneity in a key component of the brain-skull interface for the first time.

Hydraulic resistance of periarterial spaces in the brain.

BACKGROUND: Periarterial spaces (PASs) are annular channels that surround arteries in the brain and contain cerebrospinal fluid (CSF): a flow of CSF in these channels is thought to be an important part of the brain's system for clearing metabolic wastes. In vivo observations reveal that they are not concentric, circular annuli, however: the outer boundaries are often oblate, and the arteries that form the inner boundaries are often offset from the central axis. METHODS: We model PAS cross-sections as circles surrounded by ellipses and vary the radii of the circles, major and minor axes of the ellipses, and two-dimensional eccentricities of the circles with respect to the ellipses. For each shape, we solve the governing Navier-Stokes equation to determine the velocity profile for steady laminar flow and then compute the corresponding hydraulic resistance. RESULTS: We find that the observed shapes of PASs have lower hydraulic resistance than concentric, circular annuli of the same size, and therefore allow faster, more efficient flow of cerebrospinal fluid. We find that the minimum hydraulic resistance (and therefore maximum flow rate) for a given PAS cross-sectional area occurs when the ellipse is elongated and intersects the circle, dividing the PAS into two lobes, as is common around pial arteries. We also find that if both the inner and outer boundaries are nearly circular, the minimum hydraulic resistance occurs when the eccentricity is large, as is common around penetrating arteries. CONCLUSIONS: The concentric circular annulus assumed in recent studies is not a good model of the shape of actual PASs observed in vivo, and it greatly overestimates the hydraulic resistance of the PAS. Our parameterization can be used to incorporate more realistic resistances into hydraulic network models of flow of cerebrospinal fluid in the brain. Our results demonstrate that actual shapes observed in vivo are nearly optimal, in the sense of offering the least hydraulic resistance. This optimization may well represent an evolutionary adaptation that maximizes clearance of metabolic waste from the brain.

Dependence of resting-state fMRI fluctuation amplitudes on cerebral cortical orientation relative to the direction of B0 and anatomical axes.

Functional magnetic resonance imaging (fMRI) is now capable of sub-millimetre scale measurements over the entire human brain, however with such high resolutions each voxel is influenced by the local fine-scale details of the cerebral cortical vascular anatomy. The cortical vasculature is structured with the pial vessels lying tangentially along the grey matter surface, intracortical diving arterioles and ascending venules running perpendicularly to the surface, and a randomly oriented capillary network within the parenchyma. It is well-known that the amplitude of the blood-oxygenation level dependent (BOLD) signal emanating from a vessel depends on its orientation relative to the B0-field. Thus the vascular geometric hierarchy will impart an orientation dependence to the BOLD signal amplitudes and amplitude differences due to orientation differences constitute a bias for interpreting neuronal activity. Here, we demonstrate a clear effect of cortical orientation to B0 in the resting-state BOLD-fMRI amplitude (quantified as the coefficient of temporal signal variation) for 1.1 mm isotropic data at 7T and 2 mm isotropic at 3T. The maximum bias, i.e. the fluctuation amplitude difference between regions where cortex is perpendicular to vs. parallel to B0, is about +70% at the pial surface at 7T and +11% at 3T. The B0 orientation bias declines with cortical depth, becomes progressively smaller closer to the white matter surface, but then increases again to a local maximum within the white matter just beneath the cortical grey matter, suggesting a distinct tangential network of white matter vessels that also generate a BOLD orientation effect. We further found significant (negative) biases with the cortex orientation to the anterior-posterior anatomical axis of the head: a maximum negative bias of about -30% at the pial surface at 7T and about -13% at 3T. The amount of signal variance explained by the low frequency drift, motion and the respiratory cycle also showed a cortical orientation dependence; only the cardiac cycle induced signal variance was independent of cortical orientation, suggesting that the cardiac induced component of the image time-series fluctuations is not related to a significant change in susceptibility. Although these orientation effects represent a signal bias, and are likely to be a nuisance in high-resolution analyses, they may help characterize the vascular influences on candidate fMRI acquisitions and, thereby, may be exploited to improve the neuronal specificity of fMRI.

Vascular Compartmentalization of Functional Hyperemia from the Synapse to the Pia.

Functional hyperemia, a regional increase of blood flow triggered by local neural activation, is used to map brain activity in health and disease. However, the spatial-temporal dynamics of functional hyperemia remain unclear. Two-photon imaging of the entire vascular arbor in NG2-creERT2;GCaMP6f mice shows that local synaptic activation, measured via oligodendrocyte precursor cell (OPC) Ca2+ signaling, generates a synchronous Ca2+ drop in pericytes and smooth muscle cells (SMCs) enwrapping all upstream vessels feeding the activated synapses. Surprisingly, the onset timing, direction, and amplitude of vessel diameter and blood velocity changes vary dramatically from juxta-synaptic capillaries back to the pial arteriole. These results establish a precise spatial-temporal sequence of vascular changes triggered by neural activity and essential for the interpretation of blood-flow-based imaging techniques such as BOLD-fMRI.

Activation of pial and dural macrophages and dendritic cells by cortical spreading depression.

OBJECTIVE: Cortical spreading depression (CSD) has long been implicated in migraine attacks with aura. The process by which CSD, a cortical event that occurs within the blood-brain barrier (BBB), results in nociceptor activation outside the BBB is likely mediated by multiple molecules and cells. The objective of this study was to determine whether CSD activates immune cells inside the BBB (pia), outside the BBB (dura), or in both, and if so, when. METHODS: Investigating cellular events in the meninges shortly after CSD, we used in vivo two-photon imaging to identify changes in macrophages and dendritic cells (DCs) that reside in the pia, arachnoid, and dura and their anatomical relationship to TRPV1 axons. RESULTS: We found that activated meningeal macrophages retract their processes and become circular, and that activated meningeal DCs stop migrating. We found that CSD activates pial macrophages instantaneously, pial, subarachnoid, and dural DCs 6-12 minutes later, and dural macrophages 20 minutes later. Dural macrophages and DCs can appear in close proximity to TRPV1-positive axons. INTERPRETATION: The findings suggest that activation of pial macrophages may be more relevant to cases where aura and migraine begin simultaneously, that activation of dural macrophages may be more relevant to cases where headache begins 20 to 30 minutes after aura, and that activation of dural macrophages may be mediated by activation of migratory DCs in the subarachnoid space and dura. The anatomical relationship between TRPV1-positive meningeal nociceptors, and dural macrophages and DCs supports a role for these immune cells in the modulation of head pain. Ann Neurol 2018;83:508-521.

Effect of Transplantation of Mesenchymal Stem Cells on the Density of Pial Microvascular Network in Spontaneously Hypertensive Rats of Different Age.

Using a TV device for studying microcirculation (×40), we analyzed the density of the whole microvascular network and the density of arterioles in the pia mater of the sensorimotor cortex in SHR rats of different ages (3-4 and 12 months) after intracerebral transplantation of human mesenchymal stem cells. We found that the density of pial microvascular network in SHR rats receiving transplantation of human mesenchymal stem cells increased to a level observed in young Wistar-Kyoto rats.

Pia Mater Significantly Contributes to Spinal Cord Intraparenchymal Pressure in a Simulated Model of Edema.

STUDY DESIGN: Intraparenchymal pressure (IPP) measurements in an in vitro cadaveric model of CNS edema. OBJECTIVE: To assess the contribution of pia mater to IPP and the effect of piotomy. SUMMARY OF BACKGROUND DATA: Multicenter randomized control trials have shown that decompression with durotomy/duroplasty significantly decreases intracranial pressure (ICP). There is a paucity of evidence regarding the effectiveness of decompression of the spinal cord by piotomy. METHODS: The supratentorial brain and spinal cord were removed from six fresh cadavers. Dura and arachnoid mater were removed. ICP monitors were placed bilaterally in the frontal and parietal lobes, and centrally in the cervical and thoracic spinal cord. To simulate edema, specimens were submerged in hypotonic solution. IPP was recorded for 5 days. A complete dorsal midline piotomy was performed on the spinal cord and resulting IPP was recorded. RESULTS: Brain and spinal cord both increased in weight. IPP significantly increased in both brain and spinal cord. The IPP increase within the spinal cord was substantially greater (averages: all four lobes = 4.0 mm Hg; cervical = 73.7 mm Hg; thoracic = 49.3 mm Hg). After piotomy, cervical and thoracic spinal cord IPP decreased immediately (avg. postpiotomy IPP = 9.7 and 10.3, respectively). CONCLUSION: There were differential effects on brain and spinal cord IPP. Brain IPP increased only slightly, possibly because of the absence of the cranium and dura mater. In contrast, spinal cord IPP increased substantially even in the absence of the laminae, dura, and arachnoid mater. Piotomy immediately and dramatically reduced spinal cord IPP. These data are consistent with the hypothesis that spinal cord IPP is primarily dependent on constraints imposed by the pia mater. Conversely, in the absence of the cranium and dura mater, the sulci may permit the pia-invested brain to better accommodate edema without significant increases in IPP. LEVEL OF EVIDENCE: N/A.

Intracerebroventricular application of S100B selectively impairs pial arteriolar dilating function in rats.

S100B is an astrocyte-derived protein that can act through the receptor for advanced glycation endproducts (RAGE) to mediate either "trophic" or "toxic" responses. Its levels increase in many neurological conditions with associated microvascular dysregulation, such as subarachnoid hemorrhage (SAH) and traumatic brain injury. The role of S100B in the pathogenesis of microvasculopathy has not been addressed. This study was designed to examine whether S100B alters pial arteriolar vasodilating function. Rats were randomized to receive (1) artificial cerebrospinal fluid (aCSF), (2) exogenous S100B, and (3) exogenous S100B+the decoy soluble RAGE (sRAGE). S100B was infused intracerebroventricularly (icv) using an osmotic pump and its levels in the CSF were adjusted to achieve a concentration similar to what we observed in SAH. After 48 h of continuous icv infusion, a cranial window/intravital microscopy was applied to animals for evaluation of pial arteriolar dilating responses to sciatic nerve stimulation (SNS), hypercapnia, and topical suffusion of vasodilators including acetylcholine (ACh), s-nitroso-N-acetyl penicillamine (SNAP), or adenosine (ADO). Pial arteriolar dilating responses were calculated as the percentage change of arteriolar diameter in relation to baseline. The continuous S100B infusion for 48 h was associated with reduced responses to the neuronal-dependent vasodilator SNS (p<0.05) and the endothelial-dependent vasodilator ACh (p<0.05), compared to controls. The inhibitory effects of S100B were prevented by sRAGE. On the other hand, S100B did not alter the responses elicited by vascular smooth muscle cell-dependent vasodilators, namely hypercapnia, SNAP, or ADO. These findings indicate that S100B regulates neuronal and endothelial dependent cerebral arteriolar dilation and suggest that this phenomenon is mediated through RAGE-associated pathways.

Poro-elastic modeling of Syringomyelia - a systematic study of the effects of pia mater, central canal, median fissure, white and gray matter on pressure wave propagation and fluid movement within the cervical spinal cord.

Syringomyelia, fluid-filled cavities within the spinal cord, occurs frequently in association with a Chiari I malformation and produces some of its most severe neurological symptoms. The exact mechanism causing syringomyelia remains unknown. Since syringomyelia occurs frequently in association with obstructed cerebrospinal fluid (CSF) flow, it has been hypothesized that syrinx formation is mechanically driven. In this study we model the spinal cord tissue either as a poro-elastic medium or as a solid linear elastic medium, and simulate the propagation of pressure waves through an anatomically plausible 3D geometry, with boundary conditions based on in vivo CSF pressure measurements. Then various anatomic and tissue properties are modified, resulting in a total of 11 variations of the model that are compared. The results show that an open segment of the central canal and a stiff pia (relative to the cord) both increase the radial pressure gradients and enhance interstitial fluid flow in the central canal. The anterior median fissure, anisotropic permeability of the white matter, and Poisson ratio play minor roles.

Neonatal pial surface electroporation.

Over the past several years the pial surface has been identified as a germinal niche of importance during embryonic, perinatal and adult neuro- and gliogenesis, including after injury. However, methods for genetically interrogating these progenitor populations and tracking their lineages had been limited owing to a lack of specificity or time consuming production of viruses. Thus, progress in this region has been relatively slow with only a handful of investigations of this location. Electroporation has been used for over a decade to study neural stem cell properties in the embryo, and more recently in the postnatal brain. Here we describe an efficient, rapid, and simple technique for the genetic manipulation of pial surface progenitors based on an adapted electroporation approach. Pial surface electroporation allows for facile genetic labeling and manipulation of these progenitors, thus representing a time-saving and economical approach for studying these cells.

Constitutive modeling of pia-arachnoid complex.

The pia-arachnoid complex (PAC) covering the brain plays an important role in the mechanical response of the brain during impact or inertial loading. Recent studies have revealed the complicated material behavior of the PAC. In this study, the nonlinear viscoelastic, transversely isotropic material properties of the PAC were modeled as Mooney-Rivlin ground substance with collagen fibers strengthening within the meningeal plane through an exponential model. The material constants needed were determined using experimental data from in-plane tension, normal traction, and shear tests conducted on bovine specimens. Results from this study provide essential information to properly model the PAC membrane, an important component in the skull/brain interface, in a computational brain model. Such an improved representation of the skull/brain interface will enhance the accuracy of finite element models used in brain injury mechanism studies under various loading conditions.

Ovine tests of a novel spinal cord neuromodulator and dentate ligament fixation method.

BACKGROUND: To improve methods for the treatment of intractable pain, we are developing a novel intradural spinal cord stimulator that could be either attached to the dentate ligaments of the human spinal cord or fitted around the dorsal arc of the cord itself. PURPOSE: Our goal was to carry out the first in vivo tests of these attachment methods in an ovine model using custom-built devices and instrumentation. For eventual translational studies, we also explored methods of mimicking a human dentate ligament attachment technique in this large animal model. METHODS: As a starting point, we investigated details of the gross and histological anatomy of the ovine denticulate ligaments, and compared them with their human counterpart. The gap between the dura and the spinal cord in the sheep is small; hence, the denticulate ligaments are not long enough to accommodate human-scaled attachment clips. Therefore, lateral strips of the spinal-canal dura were fashioned to serve this same device attachment function. RESULTS: This form of dural anchoring was implemented surgically for fixation of a silicone membrane implant that had 12 electrodes, and somatosensory evoked potentials were obtained successfully when stimuli were applied to it. The dorsal arc clamping technique was also implemented. CONCLUSIONS: We demonstrated that the dural attachment method is an effective surrogate model for testing the human dentate ligament device fixation technique, and that this mode of fixation was preferable to dorsal arc attachment. The relevant surgical innovations, anatomical findings, and the preliminary electrophysiological data from a pial surface stimulator attached in this way are presented.

Rapid genetic targeting of pial surface neural progenitors and immature neurons by neonatal electroporation.

BACKGROUND: Recent findings have indicated the presence of a progenitor domain at the marginal zone/layer 1 of the cerebral cortex, and it has been suggested that these progenitors have neurogenic and gliogenic potential. However, their contribution to the histogenesis of the cortex remains poorly understood due to difficulties associated with genetically manipulating these unique cells in a population-specific manner. RESULTS: We have adapted the electroporation technique to target pial surface cells for rapid genetic manipulation at postnatal day 2. In vivo data show that most of these cells proliferate and progressively differentiate into both neuronal and glial subtypes. Furthermore, these cells localize to the superficial layers of the optic tectum and cerebral cortex prior to migration away from the surface. CONCLUSIONS: We provide a foundation upon which future studies can begin to elucidate the molecular controls governing neural progenitor fate, migration, differentiation, and contribution to cortical and tectal histogenesis. Furthermore, specific genetic targeting of such neural progenitor populations will likely be of future clinical interest.

Multi-echo fMRI of the cortical laminae in humans at 7 T.

Recent developments in ultra high field MRI and receiver coil technology have opened up the possibility of laminar fMRI in humans. This could offer greater insight into human brain function by elucidating both the interaction between brain regions on the basis of laminar activation patterns associated with input and output, and the interactions between laminae in a specific region. We used very high isotropic spatial resolution (0.75 mm voxel size), multi-echo acquisition (gradient-echo) in a 7 T fMRI study of human primary visual cortex (V1) and novel data analysis techniques to quantitatively investigate the echo time dependence of laminar profiles, laminar activation, and physiological noise distributions over an extended region of cortex. We found T(2)* profiles to be explicable in terms of variations in myelin content. Laminar activation profiles vary with echo time (TE): at short TE the highest signal changes are measured at the pial surface; this maximum shifts into grey matter at longer TEs. The top layers peak latest as these have the longest transverse relaxation time. Theoretical simulations and experiment suggest that the intravascular contribution to functional signal changes is significant even at long TE. Based on a temporal noise analysis we argue that the (physiological) noise contributions will ameliorate differences in sensitivity between the layers in a statistical analysis, and correlates with laminar blood volume distribution. We also show that even at this high spatial resolution the physiological noise limit to sensitivity is reached within V1, implying that cortical sub-regions can be examined with this technique.