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Background: Cerebrospinal fluid (CSF) circulation in the brain has garnered considerable attention in recent times. In contrast, there have been fewer studies focused on the spine, despite the expected importance of CSF circulation in disorders specific to the spine, including syringomyelia. The driving forces that regulate spinal CSF flow are not well defined and are likely to be different to the brain given the anatomical differences and proximity to the heart and lungs. The aims of this study were to determine the effects of heart rate, blood pressure and respiration on the distribution of CSF tracers in the spinal subarachnoid space, as well as into the spinal cord interstitium., Methods: In Sprague Dawley rats, physiological parameters were manipulated such that the effects of spontaneous breathing (generating alternating positive and negative intrathoracic pressures), mechanical ventilation (positive intrathoracic pressure only), tachy/bradycardia, as well as hyper/hypotension were separately studied. To investigate spinal CSF hydrodynamics, in vivo near-infrared imaging of intracisternally infused indocyanine green was performed. CSF tracer transport was further characterised with in vivo two-photon intravital imaging. Tracer influx at a microscopic level was quantitatively characterised by ex vivo epifluorescence imaging of fluorescent ovalbumin., Results: Compared to mechanically ventilated controls, spontaneous breathing animals had significantly greater movement of tracer in the subarachnoid space. There was also greater influx into the spinal cord interstitium. Hypertension and tachycardia had no significant effect on spinal subarachnoid spinal CSF tracer flux and exerted less effect than respiration on tracer influx into the spinal cord., Conclusions: Intrathoracic pressure changes that occur over the respiratory cycle, particularly decreased intrathoracic pressures generated during inspiration, have a profound effect on tracer movement after injection into spinal CSF and increase cord parenchymal tracer influx. Arterial pulsations likely drive fluid transport from perivascular spaces into the surrounding interstitium, but their overall impact is less than that of the respiratory cycle on net tracer influx., (© 2022. The Author(s).)

Background: Disruption of cerebrospinal fluid (CSF)/interstitial fluid (ISF) exchange in the spinal cord is likely to contribute to central nervous system (CNS) diseases that involve abnormal fluid accumulation, including spinal cord oedema and syringomyelia. However, the physiological factors that govern fluid transport in the spinal cord are poorly understood. The aims of this study were to determine the effects of cardiac pulsations and respiration on tracer signal increase, indicative of molecular movement following infusion into the spinal cord grey or white matter., Methods: In Sprague Dawley rats, physiological parameters were manipulated such that the effects of spontaneous breathing (generating alternating positive and negative intrathoracic pressures), mechanical ventilation (positive intrathoracic pressure only), tachycardia (heart atrial pacing), as well as hypertension (pharmacologically induced) were separately studied. Since fluid outflow from the spinal cord cannot be directly measured, we assessed the molecular movement of fluorescent ovalbumin (AFO-647), visualised by an increase in tracer signal, following injection into the cervicothoracic spinal grey or white matter., Results: Tachycardia and hypertension increased AFO-647 tracer efflux, while the concomitant negative and positive intrathoracic pressures generated during spontaneous breathing did not when compared to the positive-pressure ventilated controls. Following AFO-647 tracer injection into the spinal grey matter, increasing blood pressure and heart rate resulted in increased tracer movement away from the injection site compared to the hypotensive, bradycardic animals (hypertension: p = 0.05, tachycardia: p < 0.0001). Similarly, hypertension and tachycardia produced greater movement of AFO-647 tracer longitudinally along the spinal cord following injection into the spinal white matter (p < 0.0001 and p = 0.002, respectively). Tracer efflux was strongly associated with all blood vessel types., Conclusions: Arterial pulsations have profound effects on spinal cord interstitial fluid homeostasis, generating greater tracer efflux than intrathoracic pressure changes that occur over the respiratory cycle, demonstrated by increased craniocaudal CSF tracer movement in the spinal cord parenchyma., (© 2021. The Author(s).)

Background: Cerebrospinal fluid (CSF) is thought to flow into the brain via perivascular spaces around arteries, where it mixes with interstitial fluid. The precise details concerning fluid outflow remain controversial. Although fluid dynamics have been studied in the brain, little is known about spinal cord fluid inflow and outflow. Understanding the normal fluid physiology of the spinal cord may give insight into the pathogenesis of spinal cord oedema and CSF disorders such as syringomyelia. We therefore aimed to determine the fluid outflow pathways in the rat spinal cord., Methods: A fluorescent tracer, Alexa-Fluor ® -647 Ovalbumin, was injected into the extracellular space of either the cervicothoracic lateral white matter or the grey matter in twenty-two Sprague-Dawley rats over 250 s. The rats were sacrificed at 20 or 60 min post injection. Spinal cord segments were sectioned and labelled with vascular antibodies for immunohistochemistry., Results: Fluorescent tracer was distributed over two to three spinal levels adjacent to the injection site. In grey matter injections, tracer spread radially into the white matter. In white matter injections, tracer was confined to and redistributed along the longitudinal axonal fibres. Tracer was conducted towards the pial and ependymal surfaces along vascular structures. There was accumulation of tracer around the adventitia of the intramedullary arteries, veins and capillaries, as well as the extramedullary vessels. A distinct layer of tracer was deposited in the internal basement membrane of the tunica media of arteries. In half the grey matter injections, tracer was detected in the central canal., Conclusions: These results suggest that in the spinal cord interstitial fluid movement is modulated by tissue diffusivity of grey and white matter. The central canal, and the compartments around or within blood vessels appear to be dominant pathways for fluid drainage in these experiments. There may be regional variations in fluid outflow capacity due to vascular and other anatomical differences between the grey and white matter.

Perivascular spaces play a pivotal role in the exchange between cerebrospinal and interstitial fluids, and in the clearance of waste in the CNS, yet their precise anatomical components are not well described. The aim of this study was to characterise the ultrastructure of perivascular spaces and their role in the transport of fluid, in the spinal cord of healthy rats, using transmission electron microscopy. The distribution of cerebrospinal fluid tracers injected into the subarachnoid space was studied using light, confocal and electron microscopy. Perivascular spaces were found around arterioles and venules, but not capillaries, throughout the spinal cord white and grey matter. They contained fibroblasts and collagen fibres, and were continuous with the extracellular spaces of the surrounding tissue. At 5 min post injection, tracers were seen in the subarachnoid space, the peripheral white matter, the perivascular spaces, basement membranes, extracellular spaces of the surrounding tissue, and surprisingly, in the lumen of blood vessels, suggesting trans-vascular clearance. These findings point out an unrecognised outflow pathway for CNS fluids, with potential implications for volume regulation in health and disease states, but also clinically for the detection of CNS-derived biomarkers in plasma, the immune response and drug pharmacokinetics.

Syringomyelia (a spinal cord cyst) usually develops as a result of conditions that cause cerebrospinal fluid (CSF) obstruction. The mechanism of syrinx formation and enlargement remains unclear, though previous studies suggest that the fluid enters via the perivascular spaces (PVS) of the penetrating arteries of the spinal cord, and that alterations in the CSF pulse timing and pressure could contribute to enhanced PVS inflow. This study uses an idealised computational model of the PVS to investigate the factors that influence peri-arterial fluid flow. First, we used three sample patient-specific models to explore whether changes in subarachnoid space (SAS) pressures in individuals with and without syringomyelia could influence PVS inflow. Second we conducted a parametric study to determine how features of the CSF pulse altered perivascular fluid, including alterations to timing and magnitude of the peak SAS pressure, the timing of reversal from high to low pressure (diastolic phase), and the area under the pressure-time curve. The model for the patient with syringomyelia had higher net CSF inflow to the PVS than the two subjects without syringomyelia. In the parametric study, only increasing the area under the high pressure region of the SAS pulse substantially increased PVS inflow, when coupled with a temporal shift in arterial and SAS pulses. This suggests that a period of sustained high SAS pressure while arterial diameter is low may increase net CSF pumping into the PVS.

Object: Noncommunicating canalicular syringomyelia occurs in up to 65% of patients with Chiari malformation Type I. The pathogenesis of this type of syringomyelia is poorly understood and treatment is not always effective. Although it is generally thought that syringomyelia is simply an accumulation of CSF from the subarachnoid space, the pathogenesis is likely to be more complex and may involve cellular and molecular processes. Aquaporin-4 (AQP4) has been implicated in numerous CNS pathological conditions involving fluid accumulation, including spinal cord edema. There is evidence that AQP4 facilitates the removal of extracellular water following vasogenic edema. The aim of this study was to investigate AQP4 expression and the structural and functional integrity of the blood-spinal cord barrier (BSCB) in a model of noncommunicating canalicular syringomyelia., Methods: A kaolin-induced model of canalicular syringomyelia was used to investigate BSCB permeability and AQP4 expression in 27 adult male Sprague-Dawley rats. Control groups consisted of nonoperated, laminectomy-only, and saline-injected animals. The structural integrity of the BSCB was assessed using immunoreactivity to endothelial barrier antigen. Functional integrity of the BSCB was assessed by extravasation of systemically injected horseradish peroxidase (HRP) at 1, 3, 6, or 12 weeks after surgery. Immunofluorescence was used to assess AQP4 and glial fibrillary acidic protein (GFAP) expression at 12 weeks following syrinx induction., Results: Extravasation of HRP was evident surrounding the central canal in 11 of 15 animals injected with kaolin, and in 2 of the 5 sham-injected animals. No disruption of the BSCB was observed in laminectomy-only controls. At 12 weeks the tracer leakage was widespread, occurring at every level rostral to the kaolin injection. At this time point there was a decrease in EBA expression in the gray matter surrounding the central canal from C-5 to C-7. Aquaporin-4 was expressed in gray- and white-matter astrocytes, predominantly at the glia limitans interna and externa, and to a lesser extent around neurons and blood vessels, in both control and syrinx animals. Expression of GFAP and APQ4 directly surrounding the central canal in kaolin-injected animals was variable and not significantly different from expression in controls., Conclusions: This study demonstrated a prolonged disruption of the BSCB directly surrounding the central canal in the experimental model of noncommunicating canalicular syringomyelia. The disruption was widespread at 12 weeks, when central canal dilation was most marked. Loss of integrity of the barrier with fluid entering the interstitial space of the spinal parenchyma may contribute to enlargement of the canal and progression of syringomyelia. Significant changes in AQP4 expression were not observed in this model of canalicular syringomyelia. Further investigation is needed to elucidate whether subtle changes in AQP4 expression occur in canalicular syringomyelia.

Object: The mechanisms of syringomyelia have long puzzled neurosurgeons and researchers alike due to difficulties in identifying the driving forces behind fluid flow into a syrinx, apparently against a pressure gradient between the spinal cord and the subarachnoid space (SAS). Recently, the synchronization between CSF flow and the cardiac cycle has been postulated to affect fluid flow in the spinal cord. This study aims to determine the effect of changes in the timing of SAS pressure on perivascular flow into the spinal cord., Methods: This study uses a computational fluid dynamics model to investigate whether the relative timing of a spinal artery cardiovascular pulse wave and fluid pressure in the spinal SAS can influence CSF flow in the perivascular spaces., Results: The results show that the mass flow rate of CSF through a model periarterial space is strongly influenced by the relative timing of the arterial pulse wave and the SAS pressure., Conclusions: These findings suggest that factors that might alter the timing of the pulse wave or the fluid flow in the SAS could potentially affect fluid flow into a syrinx.

A number of studies have investigated the mechanical properties of adult spinal cord under tension, however it is not known whether age has an effect on these properties. This is of interest to those aiming to understand the clinical differences between adults and children with spinal cord injury (e.g. severity and recovery), and those developing experimental or computational models for paediatric spinal cord injury. Entire spinal cords were freshly harvested from neonatal rats (14 days) and tested in vitro under uniaxial tension at a range of strain rates (0.2, 0.02, 0.002/s) to a range of strains (2%, 3.5%, 5%), with relaxation responses being recorded for 15 min. These mechanical properties were compared to previously reported data from similar experiments on adult rat spinal cords, and the peak stress and the stress after 15 min of relaxation were found to be significantly higher for spinal cords from adults than neonates (p<0.001). A non-linear viscoelastic model was developed and was observed to adequately predict the mechanical behaviour of this tissue. The model developed in this study may be of use in computational models of paediatric spinal cord. The significant differences between adult and neonatal spinal cord properties may explain the higher initial severity of spinal cord injury in children and may have implications for the development of experimental animal models for paediatric spinal cord injury, specifically for those aiming to match the injury severity with adult experimental models.

Characterizing the mechanical properties of soft biological tissues presents a formidable challenge. In order to ensure that the structure of the specimens is at a repeatable reference state, preconditioning is commonly performed before the actual test. However, the exact mechanisms of preconditioning remain unclear and more research on this issue needs to be undertaken so that the methods for preconditioning can be refined to reduce experimental variability. Previous studies have suggested that the choice of preconditioning strain may influence the measured properties. In this study, uniaxial tests were performed on three groups of spinal cord tissues. Two groups (Groups 1 and 2) were preconditioned at 5% strain and tested to 5% and 2% each, while the third (Group 3) was preconditioned at 2% strain and tested to 2%. The peak stress measured at 2% strain for group 3 was 0.0054+/-0.0035MPa and this was significantly higher than group 2 (134%; p=0.015) and group 1 (p=0.005). However, the ratio of peak to equilibrium stress and the relaxation time were similar for both preconditioning strains. This study suggests that in future work, the preconditioning strain should be reported and the highest strain being used in the study should be used for preconditioning. This is important to allow meaningful comparison of test data within the study and also with other studies.

To prevent spinal cord injury, optimize treatments for it, and better understand spinal cord pathologies such as spondylotic myelopathy, the interaction between the spinal column and the spinal cord during injury and pathology must be understood. The spinal cord is a complex and very soft tissue that changes properties rapidly after death and is difficult to model. Our objective was to develop a physical surrogate spinal cord with material properties closely corresponding to the in vivo human spinal cord that would be suitable for studying spinal cord injury under a variety of injurious conditions. Appropriate target material properties were identified from published studies and several candidate surrogate materials were screened, under uniaxial tension, in a materials testing machine. QM Skin 30, a silicone elastomer, was identified as the most appropriate material. Spinal cords manufactured from QM Skin 30 were tested under uniaxial tension and transverse compression. Rectangular specimens of QM Skin 30 were also tested under uniform compression. QM Skin 30 produced surrogate cords with a Young's modulus in tension and compression approximately matching values reported for in vivo animal spinal cords (0.25 and 0.20 MPa, respectively). The tensile and compressive Young's modulus and the behavior of the surrogate cord simulated the nonlinear behavior of the in vivo spinal cord.

Knowledge of the biomechanical properties of central nervous system (CNS) tissues is important for understanding mechanisms and thresholds for injury, and aiding development of computer or surrogate models of these tissues. Many investigations have been conducted to estimate the properties of CNS tissues including under shear, compressive and tensile loading, however there is much variability in this body of literature, making it difficult to separate the material properties from effects that result from a given experimental protocol. This review summarises previous studies of brain and spinal cord properties; discussing their main findings and points of difference, and displays the reported data on comparable scales. Additionally, based on the observed effects of methodological choices on reported tissue properties, recommendations for future studies of brain and spinal cord properties are made.

Clinically, spinal cord injuries (SCI) in infants are different from SCIs in adults. SCI is rarer in infants, and the most common types of associated spinal column injury are different for adults and infants. Initially, infants tend to have higher injury severities and mortality; however, young survivors of SCI typically have greater and more rapid functional recovery. The objective of this study was to contrast the biomechanics and neuropathology of SCI in adult and neonatal rats to investigate these differences. Thoracolumbar vertebrae of anaesthetized rats were dislocated laterally (T12 held stationary and L1 displaced laterally, with T13 between these levels) by 10 mm at 250 mm/sec in adults and by 4 mm at 100 mm/sec in neonates (13-15 days), and rats were euthanized 6 h later. Spinal cord sections were stained to detect hemorrhage (with hematoxylin and eosin [H&E]), axonal injury (with beta-amyloid precursor protein [betaAPP]), and neuronal nuclei (with NeuN). Maximum load was significantly higher in adults (25.7 +/- 2.4N) than neonates (11.0 +/- 2.4N; p < 0.001). Adult and neonatal hemorrhage volumes were not significantly different for either the raw or normalized data sets (p = 0.064 for normalized dataset). Un-normalized axonal injury densities were similar for adults and neonates, but normalized axonal injury density was significantly higher in neonates (p < 0.001). Reduction of NeuN immunoreactivity was significantly lower in neonates, for both un-normalized (p < 0.004) and normalized (p < 0.001) data sets. The findings of this study may explain the different common types of spinal column injury associated with SCI, and the greater initial severity of SCI in infants.

Freshly excised rat spinal cords were tested in uniaxial tension, in vitro, at strain rates ranging from 0.002 to 0.2 s-1. Stress relaxation tests were performed for a range of strains from 2% to 5%, with the relaxation behaviour being recorded for a period of at least 30 min. Samples exhibited a characteristic "J" shaped non-linear stress-strain response, with stiffness increasing with applied strain. The cords were labelled with rows of small markers and the uniaxial tension tests were recorded via video. Subsequent image analysis enabled the distribution of strain on the cord surface to be determined. Viscoelastic models were developed to model the mechanical behaviour of the specimens and were found to adequately describe the material behaviour.

This study was conducted to determine whether local arterial pulsations are sufficient to cause cerebrospinal fluid (CSF) flow along perivascular spaces (PVS) within the spinal cord. A theoretical model of the perivascular space surrounding a "typical" small artery was analysed using computational fluid dynamics. Systolic pulsations were modelled as travelling waves on the arterial wall. The effects of wave geometry and variable pressure conditions on fluid flow were investigated. Arterial pulsations induce fluid movement in the PVS in the direction of arterial wave travel. Perivascular flow continues even in the presence of adverse pressure gradients of a few kilopascals. Flow rates are greater with increasing pulse wave velocities and arterial deformation, as both an absolute amplitude and as a proportion of the PVS. The model suggests that arterial pulsations are sufficient to cause fluid flow in the perivascular space even against modest adverse pressure gradients. Local increases in flow in this perivascular pumping mechanism or reduction in outflow may be important in the etiology of syringomyelia.

Posttraumatic syringomyelia (PTS) is an enigmatic condition characterized by the development of fluid-filled cysts (syrinxes) within the spinal cord. Perivascular spaces (PVS) are a critical component of fluid transport within the central nervous system (CNS), with dilated PVSs variably implicated in the pathogenesis of syringomyelia. The extent and spatial distribution of dilated PVSs in syringomyelia, however, remains unclear. This study aims to develop a method to assess PVS dimensions across multiple spinal cord segments in rats with PTS. Syrinxes were induced in two Sprague–Dawley rats at C6/7 with computer-controlled motorized spinal cord impaction; two control rats underwent sham laminectomies. Spinal cord segments were obtained at C4, C6 and C8, cleared via tissue clearing protocols, stained with immunofluorescent antibodies and imaged under confocal microscopy. Qualitative and quantitative analyses of PVS size were performed. Arteriolar PVSs were enlarged in the perisyringeal region of the spinal cord, compared to the control cord. No PVS enlargement was observed above or below the syrinx. These results confirm previous incidental findings of enlarged PVSs in the perisyringeal region, providing new insights into PVS dimensions across multiple spinal segments, and providing a novel method for quantifying spinal cord perivascular space size distributions. [ABSTRACT FROM AUTHOR]

Aquaporin-4 (AQP4) has been implicated in post-traumatic syringomyelia (PTS), a disease characterised by the formation of fluid-filled cysts in the spinal cord. This study investigated the expression of AQP4 around a mature cyst (syrinx) and the effect of pharmacomodulation of AQP4 on syrinx size. PTS was induced in male Sprague–Dawley rats by computerized spinal cord impact and subarachnoid kaolin injection. Immunofluorescence of AQP4 was carried out on mature syrinx tissue 12 weeks post-surgery. Increased AQP4 expression corresponded to larger, multiloculated cysts (R2 = 0.94), yet no localized changes to AQP4 expression in perivascular regions or the glia limitans were present. In a separate cohort of animals, at 6 weeks post-surgery, an AQP4 agonist (AqF026), antagonist (AqB050), or vehicle was administered daily over 4 days, with MRIs performed before and after the completion of treatment. Histological analysis was performed at 12 weeks post-surgery. Syrinx volume and length were not altered with AQP4 modulation. The correlation between increased AQP4 expression with syrinx area suggests that AQP4 or the glia expressing AQP4 are recruited to regulate water movement. Given this, further investigation should examine AQP4 modulation with dose regimens at earlier time-points after PTS induction, as these may alter the course of syrinx development. [ABSTRACT FROM AUTHOR]

Circulation of fluid through the central nervous system maintains fluid homeostasis and is involved in solute clearance. The glymphatic system is hypothesised to facilitate waste clearance in the brain, with inflow via periarterial spaces, bulk flow through the parenchyma, and outflow via perivenous spaces. The driving force for this mechanism is unknown. Previous modelling in the spinal cord suggests that timing offsets between arterial and subarachnoid space pressure pulses can enable net inflow in perivascular spaces (PVS). This study adapted the spinal pulse offset mechanism to the brain and simulated movement of tracer particles used in experiments. Both bulk flow and diffusive movement of tracer were simulated. Intracranial pressure pulses were applied to one end of a 300-μm-long perivascular space combined with a moving arterial wall simulating arterial pulsations. The simulations indicate the pulse offset mechanism can enable net inflow via PVS; however, it is unknown whether the temporal offset required is physiologically realistic. Increasing the positive component of the ICP (intracranial pressure) pulse increased net flow. Tracer particles driven by bulk flow reached the outlet of the PVS with a net speed of ~ 16 μm/s when the permeability was two orders of magnitude higher than values in the literature. These particles were unable to penetrate into the parenchyma in the absence of diffusion. Dispersion dominated tracer movement in the parenchyma. Further research is required to reconcile discrepancies between these results, and both experimental and computational studies. [ABSTRACT FROM AUTHOR]

Abstract: Syringomyelia is a neurological disorder characterised by high pressure fluid-filled cysts within the spinal cord. As syringomyelia is associated with abnormalities of the central nervous system that obstruct cerebrospinal fluid (CSF) flow, it is thought that changes in CSF dynamics play an important role in its pathogenesis. Using three-dimensional computational models of the spinal subarachnoid space (SAS), this study aims to determine SAS obstructions, such as arachnoiditis, change in CSF dynamics in the SAS. The geometry of the SAS was reconstructed from a series of MRI images. CSF is modelled as an incompressible Newtonian fluid with a dynamic viscosity of 1mPas. Three computational models simulated CSF flow in either the unobstructed SAS, or with the SAS obstructed by a porous region simulating dorsal or circumferential arachnoiditis. The permeability of this porous obstruction was varied for the model with dorsal arachnoiditis. The results show that arachnoiditis increases flow resistance in the SAS and this is accompanied by a modest increase in magnitude and/or shift in timing (with respect to the cardiac cycle) of the CSF pressure drop across the region of arachnoiditis. This study suggests that syrinx formation may be related to a change in temporal CSF pulse pressure dynamics. [Copyright &y& Elsevier]