Diffusion tensor imaging shows mechanism-specific differences in injury pattern and progression in rat models of acute spinal cord injury.

We investigate the ability of diffusion tensor imaging (DTI) to distinguish between three experimental rat models of spinal cord injury mechanism - contusion, dislocation, and distraction. Ex vivo DTI scans were performed on cord specimens that were preserved at different time points of the acute injury (3 hr, 24 hr, and 7 days post-injury) across all three injury mechanisms. White matter was classified as abnormal if their DTI metric was substantially different from regional values measured from a set of uninjured controls, thus allowing generation of binary "white matter damage maps" which categorizes each pixel in the DTI image as "normal" or "damaged". Damage classification was most robust using thresholds in the longitudinal diffusivity, which supports previous studies that show that longitudinal diffusivity is the most robust DTI metric in depicting damage in SCI. Furthermore, the spatial damage patterns from all subjects in the same group were consolidated into a "damage occurrence ratio map", which illustrates an average damage shape that characterizes the injury mechanism. Our analysis has yielded a dataset which highlights the differences in injury pattern due to the initial mode of mechanical injury. For example, contusion produced an initial injury that emanated radially outward from the central canal, with subsequent damage along the caudal corticospinal tract and rostral gracile fasciculus; dislocation injuries showed a high level of involvement in the lateral and ventral white matter which became less apparent by 7 days post-injury, and distraction injuries were found to be less focal and more distributed rostrocaudally. This work represents a first step in adopting the use of the primary injury mechanism as a clinical prognostic factor in SCI, which may help to inform the trialing of existing neuroprotective treatment candidates, the development of new therapies as well as personalize the management of SCI for the individual patient.

Repeatability of a Dislocation Spinal Cord Injury Model in a Rat-A High-Speed Biomechanical Analysis.

Dislocation is the most common, and severe, spinal cord injury (SCI) mechanism in humans, yet there are few preclinical models. While dislocation in the rat model has been shown to produce unique outcomes, like other closed column models it exhibits higher outcome variability. Refinement of the dislocation model will enhance the testing of neuroprotective strategies, further biomechanical understanding, and guide therapeutic decisions. The overall objective of this study is to improve biomechanical repeatability of a dislocation SCI model in the rat, through the following specific aims: (i) design new injury clamps that pivot and self-align to the vertebrae; (ii) measure intervertebral kinematics during injury using the existing and redesigned clamps; and (iii) compare relative motion at the vertebrae-clamp interface to determine which clamps provide the most rigid connection. Novel clamps that pivot and self-align were developed based on the quantitative rat vertebral anatomy. A dislocation injury was produced in 34 rats at C4/C5 using either the existing or redesigned clamps, and a high-speed X-ray device recorded the kinematics. Relative motion between the caudal clamp and C5 was significantly greater in the existing clamps compared to the redesigned clamps in dorsoventral translation and sagittal rotation. This study demonstrates that relative motions can be of magnitudes that likely affect injury outcomes. We recommend such biomechanical analyses be applied to other SCI models when repeatability is an issue. For this dislocation model, the results show the importance of using clamps that pivot and self-align to the vertebrae.

Temporal Progression of Acute Spinal Cord Injury Mechanisms in a Rat Model: Contusion, Dislocation, and Distraction.

Traumatic spinal cord injuries (SCIs) occur due to different spinal column injury patterns, including burst fracture, dislocation, and flexion-distraction. Pre-clinical studies modeling different SCI mechanisms have shown distinct histological differences between these injuries both acutely (3 h and less) and chronically (8 weeks), but there remains a temporal gap. Different rates of injury progression at specific regions of the spinal cord may provide insight into the pathologies that are initiated by specific SCI mechanisms. Therefore, the objective of this study was to evaluate the temporal progression of injury at specific tracts within the white matter, for time-points of 3 h, 24 h, and 7 days, for three distinct SCI mechanisms. In this study, 96 male Sprague Dawley rats underwent one of three SCI mechanisms: contusion, dislocation, or distraction. Animals were sacrificed at one of three times post-injury: 3 h, 24 h, or 7 days. Histological analysis using eriochrome cyanide and immunostaining for MBP, SMI-312, neurofilament-H (NF-H), and ß-III tubulin were used to characterize white matter sparing and axon and myelinated axon counts. The regions analyzed were the gracile fasciculus, cuneate fasciculus, dorsal corticospinal tract, and ventrolateral white matter. Contusion, dislocation, and distraction SCIs demonstrated distinct damage patterns that progressed differently over time. Myelinated axon counts were significantly reduced after dislocation and contusion injuries in most locations and time-points analyzed (compared with sham). This indicates early myelin damage often within 3 h. Myelinated axon counts after distraction dropped early and did not demonstrate any significant progression over the next 7 days. Important differences in white matter degeneration were identified between injury types, with distraction injuries showing the least variability across time-points These findings and the observation that white matter injury occurs early, and in many cases, without much dynamic change, highlight the importance of injury type in SCI research-both clinically and pre-clinically.

Effect of Velocity and Duration of Residual Compression in a Rat Dislocation Spinal Cord Injury Model.

Early decompression of the traumatically injured and persistently compressed spinal cord is intuitively beneficial for neurological outcome. Despite considerable pre-clinical evidence of a neurological benefit to early decompression, the effect of early surgical decompression in clinical spinal cord injury (SCI) remains less clear. The discrepancy between pre-clinical and clinical results may be due to differences between the biomechanical variables used in pre-clinical animal models and the biomechanical conditions occurring in clinical injuries. These pre-clinical variables include region of spinal cord, velocity of impact, and injury mechanism. In this study, the effect of velocity and duration of residual compression on injury severity were evaluated using a novel, rodent model of cervical dislocation SCI. Fifty-two male Sprague-Dawley rats were included in five groups: two timings of decompression (24 min, 240 min), two velocities (10 mm/sec, 500 mm/sec), and a sham group. All injuries involved a 1.45-mm dorsal dislocation of the C6 vertebra relative to C5 with subsequent residual compression of 0.8 mm. Animals were evaluated for motor function using the Martinez open field, grip strength, and grooming tests for 6 weeks post-injury. Immunohistochemistry and histology following sacrifice were conducted with counts for NeuN- and choline acetyltransferase (ChAT)-positive neurons, and length of cavitation. Behavioral testing and histological analysis revealed that injuries induced by the high velocity were consistently more severe than those induced by the low velocity, with behavioral correlations ranging between 0.46 and 0.58 (p < 0.05). Longer duration of residual compression did not produce significantly more severe injuries as measured by functional tests and histology. These findings demonstrate that the velocity of the initial traumatic impact may be a more important factor than duration of residual compression in determining SCI severity in a dislocation model of SCI.

Basic biomechanics of spinal cord injury - How injuries happen in people and how animal models have informed our understanding.

The wide variability, or heterogeneity, in human spinal cord injury is due partially to biomechanical factors. This review summarizes our current knowledge surrounding the patterns of human spinal column injury and the biomechanical factors affecting injury. The biomechanics of human spinal injury is studied most frequently with human cadaveric models and the features of the two most common injury patterns, burst fracture and fracture dislocation, are outlined. The biology of spinal cord injury is typically studied with animal models and the effects of the most relevant biomechanical factors - injury mechanism, injury velocity, and residual compression, are described. Tissue damage patterns and behavioural outcomes following dislocation or distraction injury mechanisms differ from the more commonly used contusion mechanism. The velocity of injury affects spinal cord damage, principally in the white matter. Ongoing, or residual compression after the initial impact does affect spinal cord damage, but few models exist that replicate the clinical scenario. Future research should focus on the effects of these biomechanical factors in different preclinical animal models as recent data suggests that treatment outcomes may vary between models.

Development of a traumatic cervical dislocation spinal cord injury model with residual compression in the rat.

BACKGROUND: Preclinical spinal cord injury models do not represent the wide range of biomechanical factors seen in human injuries, such as spinal level, injury mechanism, velocity of spinal cord impact, and residual compression. These factors may be responsible for differences observed between experimental and clinical study results, especially related to the controversial issue of timing of surgical decompression. NEW METHOD: Somatosensory Evoked Potentials were used to: a) characterize residual compression depths in a dislocation model, and b) evaluate the physiological effect of whether or not the spinal cord was decompressed following the initial injury, prior to the application of residual compression. Modifications to vertebral clamps and the development of a novel surgical frame allowed us to conduct surgical and injury procedures in a controlled manner without the risk of additional damage to the spinal cord. Behavioural outcomes were evaluated following varying dislocation displacements, in addition to the survivability of 4 h of residual compression following a traumatic injury. RESULTS: Residual compression immediately following the initial dislocation demonstrated significantly different electrophysiological response compared to when the residual compression was delayed. COMPARISON WITH EXISTING METHOD: There are currently no other residual compression models that utilize a dislocation injury mechanism. Many residual compression studies have demonstrated the effectiveness of early decompression, however the compression of the spinal cord is often not representative of clinical traumatic injuries. Preclinical studies typically model residual compression using a sustained force through quasi-static application, when human injuries often occur at high velocities, followed by a sustained displacement occlusion of the spinal canal. CONCLUSIONS: This study has validated several novel procedural approaches and injury parameters, and provided critical details to implement in the development of a traumatic cervical dislocation SCI model with residual compression.

A method to characterize average cervical spine ligament response based on raw data sets for implementation into injury biomechanics models.

Experimental testing on cervical spine ligaments provides important data for advanced numerical modeling and injury prediction; however, accurate characterization of individual ligament response and determination of average mechanical properties for specific ligaments has not been adequately addressed in the literature. Existing methods are limited by a number of arbitrary choices made during the curve fits that often misrepresent the characteristic shape response of the ligaments, which is important for incorporation into numerical models to produce a biofidelic response. A method was developed to represent the mechanical properties of individual ligaments using a piece-wise curve fit with first derivative continuity between adjacent regions. The method was applied to published data for cervical spine ligaments and preserved the shape response (toe, linear, and traumatic regions) up to failure, for strain rates of 0.5s(-1), 20s(-1), and 150-250s(-1), to determine the average force-displacement curves. Individual ligament coefficients of determination were 0.989 to 1.000 demonstrating excellent fit. This study produced a novel method in which a set of experimental ligament material property data exhibiting scatter was fit using a characteristic curve approach with a toe, linear, and traumatic region, as often observed in ligaments and tendons, and could be applied to other biological material data with a similar characteristic shape. The resultant average cervical spine ligament curves provide an accurate representation of the raw test data and the expected material property effects corresponding to varying deformation rates.

Strain rate dependent properties of human craniovertebral ligaments.

Craniovertebral ligaments were tested to failure under tensile loading. Ligaments tested included: transverse ligament, anterior atlanto occipital membrane, posterior atlanto occipital membrane, capsular ligaments between Skull-C1 and C1-C2, anterior atlantoaxial membrane, posterior atlantoaxial membrane and the tectorial membrane/vertical cruciate/apical/alar ligament complex. The objective of this study was to obtain mechanical properties of craniovertebral ligaments of a younger population, at varying strain rates representative of automotive crash scenarios, and investigate rate and gender effects for use in numerical models of the cervical spine. There have been few studies conducted on the mechanical properties of human craniovertebral ligaments. Only one study has tested all of the ligaments, and previous studies use older age specimens (mean age 67, from most complete study). Further, tests were often not performed at elongation rates representative of car crash scenarios. Previous studies did not perform tests in an environment resembling in vivo conditions, which has been shown to have a significant effect on ligament tensile behaviour. Fifty-four craniovertebral ligaments were isolated from twenty-one spines, and tested to failure in tension under simulated in vivo temperature and hydration levels, at quasi-static (0.5 s(-1)) and high strain rates (150 s(-1)). Values for failure force, failure elongation, stiffness, and toe region elongation were obtained from force-displacement curves. Values were analyzed for strain rate and gender effects. Increased strain rate produced several significant effects including: higher failure forces for the transverse ligament and capsular ligament (Skull-C1), lower failure elongation for the tectorial membrane complex, higher stiffness for the tectorial membrane complex and capsular ligament (Skull-C1), and lower toe region elongation for capsular ligament (Skull-C1). Gender effects were limited. Ligament tests demonstrated expected rate effects. Younger specimens had a higher failure force and stiffness and failed at lower elongations than older specimens from previous studies. Gender effects suggest there may be a difference between male and female properties, but require further testing to establish greater significance.

Histological effects of residual compression sustained for 60 minutes at different depths in a novel rat spinal cord injury contusion model.

Spinal cord injuries are frequently accompanied by persisting residual compression of the spinal cord; however, it remains controversial as to what effect the sustained compression has on neurological damage. The objective of this study was to determine the influence of post-traumatic residual spinal cord compression on the extent and progression of tissue damage within a dynamic thoracic contusion rat model. Twenty-nine male Wistar rats were distributed into one of four groups: spinal cord contusion only, contusion with 40% residual compression, contusion with 90% residual compression, and a surgical control group. A moderate injury was performed (1 mm, 700 mm/sec) with our custom University of British Columbia (UBC) multimechanism apparatus, and the residual compression groups had the impactor tip maintained at 40% or 90% of the initial impact depth (1.0 mm) for 60 min post-injury. All animals were killed at 3 h post-injury, when the spinal cord was harvested and stained for hemorrhage, neuronal damage in the gray matter, and axonal disruption in the white matter. The initial contusion injury immediately damaged tissue beneath the impactor as evidenced by rapid relaxation of the reaction force on the spinal cord during the subsequent compression. Importantly, the rostral-caudal extent of intramedullary hemorrhage was 66% larger after 90% residual spinal cord compression compared to the 40% group (p=0.016). Similarly, the extent of neuronal nuclei lost in different gray matter regions was 60-86% greater after 90% residual compression compared with 40% (p<0.001). Thus, a high level of residual compression of the spinal cord following a moderate contusion injury has the potential to adversely increase the extent of tissue damage, whereas a lower level of residual compression may have little to no effect.

Strain rate dependent properties of younger human cervical spine ligaments.

The cervical spine ligaments play an essential role in limiting the physiological ranges of motion in the neck; however, traumatic loading such as that experienced in automotive crash scenarios can lead to ligament damage and result in neck injury. The development of detailed neck models to evaluate the response and the potential for injury requires accurate ligament mechanical properties at relevant loading rates. The objective of this study was to measure the mechanical properties of the cervical spine ligaments, by performing tensile tests at elongation rates relevant to car crash scenarios, using younger specimens (≤50 years), in simulated in vivo conditions, and to provide a comprehensive investigation of gender and spinal level effects. The five ligaments investigated were the anterior longitudinal ligament, posterior longitudinal ligament, capsular ligament, ligamentum flavum, and interspinous ligament. Ligaments were tested in tension at quasi-static (0.5 s(-1)), medium (20 s(-1)) and high (150-250 s(-1)) strain rates. The high strain rates represented typical car crash scenarios as determined using an existing cervical spine finite element model. In total, 261 ligament tests were performed, with approximately even distribution within elongation rate, spinal level, and gender. The measured force-displacement data followed expected trends compared to previous studies. The younger ligaments investigated in this study demonstrated less scatter, and were both stiffer and stronger than comparable data from older specimens reported in previous studies. Strain rate effects were most significant, while spinal level effects were limited. Gender effects were not significant, but consistent trends were identified, with male ligaments having a higher stiffness and failure force than female ligaments.