Contribution of vertebral [corrected] bodies, endplates, and intervertebral discs to the compression creep of spinal motion segments

Spinal segments show non-linear behavior under axial compression. It is unclear to what extent this behavior is attributable to the different components of the segment. In this study, we quantified the separate contributions of vertebral bodies and intervertebral discs to creep of a segment. Secondly, we investigated the contribution of bone and osteochondral endplate (endplates including cartilage) to the deformation of the vertebral body. From eight porcine spines a motion segment, a disc and a vertebral body were dissected and subjected to mechanical testing. In an additional test, cylindrical samples, machined from the lowest thoracic vertebrae of 11 porcine spines, were used to compare the deformation of vertebral bone and endplate. All specimens were subjected to three loading cycles, each comprising a loading phase (2.0 MPa, 15 min) and a recovery phase (0.001 MPa, 30 min). All specimens displayed substantial time-dependent height changes. Average creep was the largest in motion segments and smallest in vertebral bodies. Bone samples with endplates displayed substantially more creep than samples without. In the early phase, behavior of the vertebra was similar to that of the disc. Visco-elastic deformation of the endplate therefore appeared dominant. In the late creep phase, behavior of the segment was similar to that of isolated discs, suggesting that in this phase the disc dominated creep behavior, possibly by fluid flow from the nucleus. We conclude that creep deformation of vertebral bodies contributes substantially to creep of motion segments and that within a vertebral body endplates play a major role.     

Disc mechanics with trans-endplate partial nucleotomy are not fully restored following cyclic compressive loading and unloaded recovery

Mechanical function of the intervertebral disc is maintained through the interaction between the hydrated nucleus pulposus, the surrounding annulus fibrosus, and the superior and inferior endplates. In disc degeneration the normal transfer of load between disc substructures is compromised. The objective of this study was to explore the mechanical role of the nucleus pulposus in support of axial compressive loads over time. This was achieved by measuring the elastic slow ramp and viscoelastic stress-relaxation mechanical behaviors of cadaveric sheep motion segments before and after partial nucleotomy through the endplate (keeping the annulus fibrosus intact). Mechanics were evaluated at five conditions: Intact, intact after 10,000 cycles of compression, acutely after nucleotomy, following nucleotomy and 10,000 cycles of compression, and following unloaded recovery. Radiographs and magnetic resonance images were obtained to examine structure. Only the short time constant of the stress relaxation was altered due to nucleotomy. In contrast, cyclic loading resulted in significant and large changes to both the stiffness and stress relaxation behaviors. Moreover, the nucleotomy had little to no effect on the disc mechanics after cyclic loading, as there were no significant differences comparing mechanics after cyclic loading with or without the nucleotomy. Following unloaded recovery the mechanical changes that had occurred as a consequence of cyclic loading were restored, leaving only a sustained change in the short time constant due to the trans-endplate nucleotomy. Thus the swelling and redistribution of the remaining nucleus pulposus was not able to fully restore mechanical behaviors. This study reveals insights into the role of the nucleus pulposus in disc function, and provides new information toward the potential role of altered nucleus pulpous function in the degenerative cascade.     

Mechanisms of initial endplate failure in the human vertebral body

Endplate failure occurs frequently in osteoporotic vertebral fractures and may be related to the development of high tensile strain. To determine whether the highest tensile strains in the vertebra occur in the endplates, and whether such high tensile strains are associated with the material behavior of the intervertebral disc, we used micro-CT-based finite element analysis to assess tissue-level strains in 22 elderly human vertebrae (81.5±9.6 years) that were compressed through simulated intervertebral discs. In each vertebra, we compared the highest tensile and compressive strains across the different compartments: endplates, cortical shell, and trabecular bone. The influence of Poisson-type expansion of the disc on the results was determined by compressing the vertebrae a second time in which we suppressed the Poisson expansion. We found that the highest tensile strains occurred within the endplates whereas the highest compressive strains occurred within the trabecular bone. The ratio of strain to assumed tissue-level yield strain was the highest for the endplates, indicating that the endplates had the greatest risk of initial failure. Suppressing the Poisson expansion of the disc decreased the amount of highly tensile-strained tissue in the endplates by 79.4±11.3%. These results indicate that the endplates are at the greatest risk of initial failure due to the development of high tensile strains, and that such high tensile strains are associated with the Poisson expansion of the disc. We conclude that initial failure of the vertebra is associated with high tensile strains in the endplates, which in turn are influenced by the material behavior of the disc.     

Fusion of the cervical vertebrae of mammals

Morphogenesis of the cervical vertebrae has been investigated in Dipis sagitt. and in Rattus norvegicus. The main distinctive feature of the Dipis embryos at the mesenchymal stage is their very thin perichordal intervertebral rings. As a result, short cartilaginous vertebral bodies and thin intervertebral discs develop, cervical segments lengthen more slowly than those of the Rattus. Because of the small length of the Dipis cervical segments, the cartilaginous neural arches and the transverse processes of 2-6 vertebrae draw nearer and fuse. Owing to the insufficient development of the Dipis intervertebral discs and the nuclei pulposae, the normal formation of the vertebral epiphyses is disturbed, this results in fusion of the neighbouring osseous vertebral bodies.     

Computational study of the role of fluid content and flow on the lumbar disc response in cyclic compression: Replication of in vitro and in vivo conditions

The intervertebral disc viscoelastic response is governed primarily by its fluid content and flow. In vivo measurements demonstrate that the disc volume, fluid content, height and nucleus pressure completely recover during resting even after diurnal loading with twice longer duration (16 vs. 8 h). In view of much longer periods required for the recovery of disc height and pressure in vitro, concerns have been raised on the fluid inflow through the endplates that might be hampered by clogged blood vessels post mortem. This in silico study aimed to identify fluid-flow dependent response of discs and conditions essential to replicate in vitro and in vivo observations.An osmo-poroelastic finite element model of the human lumbar L4-L5 disc-bone unit was used. Simulating earlier in vitro experiments on bovine discs, the loading protocol started with 8 h preload at 0.06 MPa followed by 30 high/low compression loading cycles each lasting 7.5 min at 0.5/0.06 MPa, respectively. Three different endplate configurations were investigated: free in- and outflow, no inflow and closed endplates with no flow. Additionally, the preload magnitude was increased from 0.06 MPa to 0.28 MPa and 0.50 MPa, or the initial nucleus hydration was reduced from 83% to 50%.For 0.06 MPa preload, the model with no inflow best matched in vitro trends. The model with free inflow increased segment height and nucleus pressure while the model with no fluid inflow resulted in a relatively small recovery in segment height and a rather constant nucleus pressure during unloading periods.Results highlight an excessive mobile fluid content as well as a restricted fluid inflow through endplates as likely causes of the discrepancies between in vivo and in vitro studies. To replicate in vivo conditions in vitro and in silico, disc hydration level should be controlled by adequate selection of preload magnitude/period and/or mobile fluid porosity.     

Intervertebral disc recovery after dynamic or static loading in vitro: Is there a role for the endplate?

In vivo studies on disc mechanics show loss of fluid from the intervertebral disc (IVD) during loading and full recovery during rest. Previous work indicated that in vitro recovery is hampered after static loading. The aim of the present study was to investigate the role of the endplate after dynamic and static loading on mechanical recovery in vitro. Lumbar spines (caprine) were obtained from the local slaughterhouse and stored frozen. Twenty-four intervertebral discs were thawed and subjected to a compression test in a saline bath (37 degrees C). The discs were pre-loaded at 20 N for 15 min. Three 15-min loading cycles (static: 2.0 MPa or dynamic: average load 2.0 MPa at 0.5 Hz) were applied, each followed by a 30-min period of unloading (20 N). After this protocol, the endplates of half of the discs were blocked with silicone paste and the long-term recovery protocol was applied; the discs were subjected to a single loading cycle (15 min of static or dynamic loading) followed by 10h of unloading at 20 N. All specimens showed a net loss of height and a gain in stiffness during the first part of the test. Eventually, height and stiffness were restored during a long-term recovery test. The difference in recovery between blocked and free endplates was marginal. If fluid flow plays a role during recovery in vitro, the role of the endplate appears to be limited. Our findings show no influence of loading type on recovery in vitro.     

Mechanical response of a simple finite element model of the intervertebral disc under complex loading

A simple axisymmetric finite element model of a human spine segment containing two adjacent vertebrae and the intervening intervertebral disc was constructed. The bodies and disc were modeled by three substructures; one to represent each of the vertebral bodies, the annulus fibrosus, and the nucleus pulposus. A semi-analytic technique was used to maintain the computational economies of a two-dimensional analysis when non- axisymmetric loads were imposed on the model. The response of the model to compression, shear, torsion and bending loads applied to the superior vertebral body was examined to determine the effects of disc geometry and material properties on response. Comparisons of model responses with experimentally measured responses were made to estimate material property values for which model behaviors are in agreement with measured behaviors.     

Biomechanical implications of lumbar spinal ligament transection

Many lumbar spine surgeries either intentionally or inadvertently damage or transect spinal ligaments. The purpose of this work was to quantify the previously unknown biomechanical consequences of isolated spinal ligament transection on the remaining spinal ligaments (stress transfer), vertebrae (bone remodelling stimulus) and intervertebral discs (disc pressure) of the lumbar spine. A finite element model of the full lumbar spine was developed and validated against experimental data and tested in the primary modes of spinal motion in the intact condition. Once a ligament was removed, stress increased in the remaining spinal ligaments and changes occurred in vertebral strain energy, but disc pressure remained similar. All major biomechanical changes occurred at the same spinal level as the transected ligament, with minor changes at adjacent levels. This work demonstrates that iatrogenic damage to spinal ligaments disturbs the load sharing within the spinal ligament network and may induce significant clinically relevant changes in the spinal motion segment.     

An accurate finite element model of the cervical spine under quasi-static loading

Cervical disc injury due to impact has been observed in clinical and biomechanical investigations; however, there is a lack of data that helps to elucidate the mechanisms of disc injury during these collisions. Therefore, it is necessary to understand the behavior of the cervical spine under different types of loading situations. A three dimensional finite element (FE) model for the multi-level cervical spine segment (C0-C7) was developed using computed tomography (CT) data and applied to study the internal stresses and strains of the intervertebral discs under quasi-static loading conditions. The intervertebral discs were treated as nonlinear, anisotropic and incompressible subjected to large deformations. The model accuracy was validated by comparing it with previously published experimental and numerical results for different movements. It was shown that the use of a fiber reinforced model to describe the behavior of the annulus of the discs would predict higher maximum shear strains than an isotropic one, being therefore important the use of complex constitutive models in order to be able to detect the appearance of injured zones, since those strains and stresses are supposed to be related with damage to soft tissues. Several movements were analyzed: flexion, extension and axial rotation, obtaining that the maximum shear stresses in the disc were higher for a flexo-extension movement.     

Damage accumulation location under cyclic loading in the lumbar disc shifts from inner annulus lamellae to peripheral annulus with increasing disc degeneration

It is difficult to study the breakdown of lumbar disc tissue over several years of exposure to bending and lifting by experimental methods. In our earlier published study we have shown how a finite element model of a healthy lumbar motion segment was used to predict the damage accumulation location and number of cyclic to failure under different loading conditions. The aim of the current study was to extend the continuum damage mechanics formulation to the degenerated discs and investigate the initiation and progression of mechanical damage. Healthy disc model was modified to represent degenerative discs (Thompson grade III and IV) by incorporating both geometrical and biochemical changes due to degeneration. Analyses predicted decrease in the number of cycles to failure with increasing severity of disc degeneration. The study showed that the damage initiated at the posterior inner annulus adjacent to the endplates and propagated outwards towards its periphery in healthy and grade III degenerated discs. The damage accumulated preferentially in the posterior region of the annulus. However in grade IV degenerated disc damage initiated at the posterior outer periphery of the annulus and propagated circumferentially. The finite element model predictions were consistent with the infrequent occurrence of rim lesions at early age but a much higher incidence in severely degenerated discs.     

Intervertebral disc viscoelastic parameters and residual mechanics spatially quantified using a hybrid confined/in situ indentation method

With advancing age, injury, musculoskeletal pathology or a combination of these, a degenerative cascade of biomechanical, biochemical, and nutritional alterations diminish the intervertebral discs' ability to maintain its structure and function. While the biomechanics of isolated disc tissues has been investigated across this degenerative spectrum, none have attempted to retain the in situ disc-endplate morphology during compressive tissue characterization. The objective of this study was to spatially quantify the viscoelastic parameters of the intervertebral disc throughout degeneration, including the as yet unreported residual stress/strain. This required the development of a hybrid confined/in situ indentation methodology, which preserves the disc structural morphology. At four locations of the disc (anterior-AF, right and left lateral AF, and NP) stress-relaxation tests were performed using the hybrid confined/in situ indentation method, which utilizes the vertebral endplate as the porous indenter tip. This method allows the endplate to remain interwoven with the disc tissue, retaining its native orientation. Healthy disc tissue exhibited significantly higher residual stress values compared to both moderate and severe degeneration in all locations (p<0.0156). Furthermore, the equilibrium stress at 15% strain (stress relaxation) was significantly diminished with advancing disc degeneration (p<0.0241). The equilibrium viscoelastic parameters show healthy discs encounter higher forces at the same strain level, and are able to maintain this force, where degenerated discs are unable to maintain this force throughout time. This morphology-conserved method provides insight into the spatial compressive mechanical properties of the intervertebral disc across the degeneration spectrum and will aid in modeling these tissue changes.     

Analyzing notochord segmentation and intervertebral disc formation using the twhh:gfp transgenic zebrafish model

To characterize the process of vertebral segmentation and disc formation in living animals, we analyzed tiggy-winkle hedgehog (twhh):green fluorescent protein (gfp) and sonic hedgehog (shh):gfp transgenic zebrafish models that display notochord-specific GFP expression. We found that they showed distinct patterns of expression in the intervertebral discs of late stage fish larvae and adult zebrafish. A segmented pattern of GFP expression was detected in the intervertebral disc of twhh:gfp transgenic fish. In contrast, little GFP expression was found in the intervertebral disc of shh:gfp transgenic fish. Treating twhh:gfp transgenic zebrafish larvae with exogenous retinoic acid (RA), a teratogenic factor on normal development, resulted in disruption of notochord segmentation and formation of oversized vertebrae. Histological analysis revealed that the oversized vertebrae are likely due to vertebral fusion. These studies demonstrate that the twhh:gfp transgenic zebrafish is a useful model for studying vertebral segmentation and disc formation, and moreover, that RA signaling may play a role in this process.     

Immunolocalization of type X collagen in human lumbar intervertebral discs during ageing and degeneration

 Type X collagen has so far not been reported to occur in human intervertebral discs. The objective of this study was therefore to investigate the occurrence of type X collagen in human lumbar intervertebral discs during ageing and degeneration. Ninety intervertebral discs with adjacent endplates were excised in toto from individuals (0–86 years) without known spinal disease and were processed for routine decalcified histology. Appropriate slices of each disc were processed for immunohistochemistry using a type-spec ific, monoclonal antibody raised against human type X collagen. Each intervertebral disc was examined for macroscopic and histomorphological features of disc degeneration. Immunohistochemically, a positive specific type X staining was observed in the hypertrophic zone of the growth plate and only in the interstitial matrix of juvenile (<2 years) nucleus pulposus. In adult discs, type X collagen could be localized in conjunction with advanced disc degeneration and first occurred in the disc matrix (i.e., pericellular region) of a 47-year-old specimen. Positive type X staining of the disc matrix was more frequently found in senile (>70 years) discs with end stages of disc degeneration. This study provides the first evidence for the occurrence of type X collagen in human lumbar intervertebral discs and it appears that type X collagen is re-expressed in late stages of disc degeneration. Accepted: 24 April 1997     

Mechanical design criteria for intervertebral disc tissue engineering

Due to the inability of current clinical practices to restore function to degenerated intervertebral discs, the arena of disc tissue engineering has received substantial attention in recent years. Despite tremendous growth and progress in this field, translation to clinical implementation has been hindered by a lack of well-defined functional benchmarks. Because successful replacement of the disc is contingent upon replication of some or all of its complex mechanical behaviors, it is critically important that disc mechanics be well characterized in order to establish discrete functional goals for tissue engineering. In this review, the key functional signatures of the intervertebral disc are discussed and used to propose a series of native tissue benchmarks to guide the development of engineered replacement tissues. These benchmarks include measures of mechanical function under tensile, compressive, and shear deformations for the disc and its substructures. In some cases, important functional measures are identified that have yet to be measured in the native tissue. Ultimately, native tissue benchmark values are compared to measurements that have been made on engineered disc tissues, identifying where functional equivalence was achieved, and where there remain opportunities for advancement. Several excellent reviews exist regarding disc composition and structure, as well as recent tissue engineering strategies; therefore this review will remain focused on the functional aspects of disc tissue engineering.     

Flexible device for vertebral body replacement

A novel vertebral prosthesis is presented. The prosthesis was developed for surgical procedures requiring the resection of a complete vertebral body and the adjacent intervertebral discs, the design objective being to develop a flexible implant that would be robust enought to withstand the in vivo stress environment of the human spine. In theory, a flexible implant should preserve a more normal range of motion and apply less stress to surrounding tissue than a rigid implant. A prototype implant was constructed so as to combine a rigid stainless steel structure with flexible silicon rubber elements in order to form an implant with static and dynamic mechanical characteristics similar to those of the anterior spinal column. Implant flexibility characteristics were determined from ex vivo stress-strain behaviour during bending and compressive creep testing. Results from the bending tests indicated good agreement for the lateral and sagittal bending characteristics in comparison with in vitro bending tests of human lumbar motion segments. Comparison of the implant compressive creep responsé with similar in vitro tests on human lumbar intervertebral discs also demonstrated similarities in the time-dependent mechanical parameters.     

MEMBRANE CHARACTERISTICS AND OSMOTIC BEHAVIOR OF ISOLATED ROD OUTER SEGMENTS

Freshly isolated frog rod outer segments are sensitive osmometers which retain their photosensitivity; their osmotic behavior reveals essentially the same light-sensitive Na⁺ influx observed electrophysiologically in the intact receptor cell. Using appropriate osmotic conditions we have examined freeze-etch replicas of freshly isolated outer segments to identify the membrane which regulates the flow of water and ions. Under isosmotic conditions we find that the disc to disc repeat distance is almost exactly twice the thickness of a disc. This ratio appears to be the same in a variety of vertebrate rod outer segments and can be reliably measured in freeze-etch images. Under all our osmotic conditions the discs appear nearly collapsed. However, when the length of the outer segment is reduced by hyperosmotic shocks the discs move closer together. This markedly reduces the ratio of repeat distance to disc thickness since disc thickness remains essentially constant. Thus, the length reduction of isolated outer segments after hyperosmotic shocks primarily results from reduction of the extradisc volume. Since the discs are free floating and since they undergo negligibly small changes in volume, the plasma membrane alone must be primarily responsible for regulating the water flux and the light-sensitive Na⁺ influx in freshly isolated outer segments. On this basis we calculate, from the osmotic behavior, that the plasma membrane of frog rod outer segment has a Na⁺ permeability constant of about 2.8 x 10^-6 cm/s and an osmotic permeability coefficient of greater than 2 x 10^-3 cm/s.     

Personalised mechanical properties of scoliotic vertebrae determined in vivo using tomodensitometry

This in vivo study investigated the mechanical properties of apical scoliotic vertebrae using computed tomography (CT) and finite element (FE) meshing. CT examination was performed on seven scoliotic girls. FE meshing of each vertebral body allowed automatic mapping of the CT scan and the visualisation of the bone density distribution. Centroids and mass centres were compared to analyse the mechanical properties distribution. Compared to the centroid, the mass centre migrated into the concavity of the curvature. The three vertebrae of a same patient had the same body migration behaviour because they were located at the curvature apex. This observation was verified in the coronal plane, but not in the sagittal plane. These results represent new data over few geometrical analyses of scoliotic vertebrae. Same in vivo personalisation of mechanical properties should be performed on intervertebral discs or ligaments to personalise stiffness properties of the spine for the biomechanical modelling of human torso. Moreover, do this mechanical deformation of scoliotic vertebrae, that appears before the vertebral wedging, could be a predictive tool in scoliosis treatment?     

Moderately degenerated lumbar motion segments: Are they truly unstable?

The two main load bearing tissues of the intervertebral disc are the nucleus pulposus and the annulus fibrosus. Both tissues are composed of the same basic components, but differ in their organization and relative amounts. With degeneration, the clear distinction between the two tissues disappears. The changes in biochemical content lead to changes in mechanical behaviour of the intervertebral disc. The aim of the current study was to investigate if well-documented moderate degeneration at the biochemical and fibre structure level leads to instability of the lumbar spine. By taking into account biochemical and ultrastructural changes to the extracellular matrix of degenerating discs, a set of constitutive material parameters were determined that described the individual tissue behaviour. These tissue biomechanical models were then used to simulate dynamic behaviour of the degenerated spinal motion segment, which showed instability in axial rotation, while a stabilizing effect in the other two principle bending directions. When a shear load was applied to the degenerated spinal motion segment, no sign of instability was found. This study found that reported changes to the nucleus pulposus and annulus fibrosus matrix during moderate degeneration lead to a more stable spinal motion segment and that such biomechanical considerations should be incorporated into the general pathophysiological understanding of disc degeneration and how its progress could affect low back pain and its treatments thereof.     

Preparation of Intact Bovine Tail Intervertebral Discs for Organ Culture

The intervertebral disc (IVD) is the joint of the spine connecting vertebra to vertebra. It functions to transmit loading of the spine and give flexibility to the spine. It composes of three compartments: the innermost nucleus pulposus (NP) encompassing by the annulus fibrosus (AF), and two cartilaginous endplates connecting the NP and AF to the vertebral body on both sides. Discogenic pain possibly caused by degenerative intervertebral disc disease (DDD) and disc herniations has been identified as a major problem in our modern society. To study possible mechanisms of IVD degeneration, in vitro organ culture systems with live disc cells are highly appealing. The in vitro culture of intact bovine coccygeal IVDs has advanced to a relevant model system, which allows the study of mechano-biological aspects in a well-controlled physiological and mechanical environment. Bovine tail IVDs can be obtained relatively easy in higher numbers and are very similar to the human lumbar IVDs with respect to cell density, cell population and dimensions. However, previous bovine caudal IVD harvesting techniques retaining cartilaginous endplates and bony endplates failed after 1-2 days of culture since the nutrition pathways were obviously blocked by clotted blood. IVDs are the biggest avascular organs, thus, the nutrients to the cells in the NP are solely dependent on diffusion via the capillary buds from the adjacent vertebral body. Presence of bone debris and clotted blood on the endplate surfaces can hinder nutrient diffusion into the center of the disc and compromise cell viability. Our group established a relatively quick protocol to "crack"-out the IVDs from the tail with a low risk for contamination. We are able to permeabilize the freshly-cut bony endplate surfaces by using a surgical jet lavage system, which removes the blood clots and cutting debris and very efficiently reopens the nutrition diffusion pathway to the center of the IVD. The presence of growth plates on both sides of the vertebral bone has to be avoided and to be removed prior to culture. In this video, we outline the crucial steps during preparation and demonstrate the key to a successful organ culture maintaining high cell viability for 14 days under free swelling culture. The culture time could be extended when appropriate mechanical environment can be maintained by using mechanical loading bioreactor. The technique demonstrated here can be extended to other animal species such as porcine, ovine and leporine caudal and lumbar IVD isolation.     

Initiation and progression of mechanical damage in the intervertebral disc under cyclic loading using continuum damage mechanics methodology: A finite element study

It is difficult to study the breakdown of disc tissue over several years of exposure to bending and lifting by experimental methods. There is also no finite element model that elucidates the failure mechanism due to repetitive loading of the lumbar motion segment. The aim of this study was to refine an already validated poro-elastic finite element model of lumbar motion segment to investigate the initiation and progression of mechanical damage in the disc under simple and complex cyclic loading conditions. Continuum damage mechanics methodology was incorporated into the finite element model to track the damage accumulation in the annulus in response to the repetitive loading. The analyses showed that the damage initiated at the posterior inner annulus adjacent to the endplates and propagated outwards towards its periphery under all loading conditions simulated. The damage accumulated preferentially in the posterior region of the annulus. The analyses also showed that the disc failure is unlikely to happen with repetitive bending in the absence of compressive load. Compressive cyclic loading with low peak load magnitude also did not create the failure of the disc. The finite element model results were consistent with the experimental and clinical observations in terms of the region of failure, magnitude of applied loads and the number of load cycles survived.