Low-back pain (LBP) is a common medical complaint and associated with high societal costs. Degeneration of the intervertebral disc (IVD) is assumed to be an important causal factor of LBP. IVDs are continuously mechanically loaded and both positive and negative effects have been attributed to different loading conditions.In order to study mechanical loading effects, degeneration-associated processes and/or potential regenerative therapies in IVDs, it is imperative to maintain the IVDs' structural integrity. While in vivo models provide comprehensive insight in IVD biology, an accompanying organ culture model can focus on a single factor, such as loading and may serve as a prescreening model to reduce life animal testing. In the current study we examined the feasibility of organ culture of caprine lumbar discs, with the hypothesis that a simulated-physiological load will optimally preserve IVD properties.Lumbar caprine IVDs (n = 175) were cultured in a bioreactor up to 21 days either without load, low dynamic load (LDL), or with simulated-physiological load (SPL). IVD stiffness was calculated from measurements of IVD loading and displacement. IVD nucleus, inner- and outer annulus were assessed for cell viability, cell density and gene expression. The extracellular matrix (ECM) was analyzed for water, glycosaminoglycan and total collagen content.IVD biomechanical properties did not change significantly with loading conditions. With SPL, cell viability, cell density and gene expression were preserved up to 21 days. Both unloaded and LDL resulted in decreased cell viability, cell density and significant changes in gene expression, yet no differences in ECM content were observed in any group.In conclusion, simulated-physiological loading preserved the native properties of caprine IVDs during a 21-day culture period. The characterization of caprine IVD response to culture in the LDCS under SPL conditions paves the way for controlled analysis of degeneration- and regeneration-associated processes in the future.

Mechanical overloading of the spine is associated with low back pain and intervertebral disc (IVD) degeneration. How excessive loading elicits degenerative changes in the IVD is poorly understood. Comprehensive knowledge of the interaction between mechanical loading, cell responses and changes in the extracellular matrix of the disc is needed in order to successfully intervene in this process. The purpose of the current study was to investigate whether dynamic and static overloading affect caprine lumbar discs differently and what mechanisms lead to mechanically induced IVD degeneration. Lumbar caprine IVDs (n = 175) were cultured 7, 14 and 21 days under simulated-physiological loading (control), high dynamic or high static loading. Axial deformation and stiffness were continuously measured. Cell viability, cell density, and gene expression were assessed in the nucleus, inner- and outer annulus. The extracellular matrix (ECM) was analyzed for water, glycosaminoglycan and collagen content. IVD height loss and changes in axial deformation were gradual with dynamic and acute with static overloading. Dynamic overloading caused cell death in all IVD regions, whereas static overloading mostly affected the outer annulus. IVDs expression of catabolic and inflammation-related genes was up-regulated directly, whereas loss of water and glycosaminoglycan were significant only after 21 days. Static and dynamic overloading both induced pathological changes to caprine lumbar IVDs within 21 days. The mechanism by which they inflict biomechanical, cellular, and extracellular changes to the nucleus and annulus differed. The described cascades provide leads for the development of new pharmacological and rehabilitative therapies to halt the progression of DDD.

Introduction

IntroductionLumbar hernias occur mostly in the posterolateral region of IVDs and mechanical loading is an important risk factor. Studies show that dynamic and static overloading affect the nucleus and annulus of the IVD differently. We hypothesize there is also variance in the effect of overloading on the IVD’s anterior, lateral and posterior annulus, which could explain the predilection of herniations in the posterolateral region. We assessed the regional mechanical and cellular responses of lumbar caprine discs to dynamic and static overloading.Material and methodsIVDs (n = 125) were cultured in a bioreactor and subjected to simulated-physiological loading (SPL), high dynamic (HD), or high static (HS) overloading. The effect of loading was determined in five disc regions: nucleus, inner-annulus and anterior, lateral and posterior outer-annulus. IVD height loss and external pressure transfer during loading were measured, cell viability was mapped and quantified, and matrix integrity was assessed.ResultsDuring culture, overloaded IVDs lost a significant amount of height, yet the distribution of axial pressure remained unchanged. HD loading caused cell death and disruption of matrix in all IVD regions, whereas HS loading particularly affected cell viability and matrix integrity in the posterior region of the outer annulus.ConclusionAxial overloading is detrimental to the lumbar IVD. Static overloading affects the posterior annulus more strongly, while the nucleus is relatively spared. Hence, static overloading predisposes the disc for posterior herniation. These findings could have implications for working conditions, in particular of sedentary occupations, and the design of interventions aimed at prevention and treatment of early intervertebral disc degeneration.

Purpose Lumbar laminectomy affects spinal stability in shear loading. However, the effects of laminectomy on torsion biomechanics are unknown. The purpose of this study was to investigate the effect of laminectomy on torsion stiffness and torsion strength of lumbar spinal segments following laminectomy and whether these biomechanical parameters are affected by disc degeneration and bone mineral density (BMD). Methods Ten human cadaveric lumbar spines were obtained (age 75.5, range 59-88). Disc degeneration (MRI) and BMD (DXA) were assessed. Disc degeneration was classified according to Pfirrmann and dichotomized in mild or severe. BMD was defined as high BMD (Cmedian BMD) or low BMD (\median BMD). Laminectomy was performed either on L2 (59) or L4 (59). Twenty motion segments (L2-L3 and L4-L5) were isolated. The effects of laminectomy, disc degeneration and BMD on torsion stiffness (TS) and torsion moments to failure (TMF) were studied.Results Load-displacement curves showed a typical bi-phasic pattern with an early torsion stiffness (ETS), late torsion stiffness (LTS) and a TMF. Following laminectomy, ETS decreased 34.1 % (p \ 0.001), LTS decreased 30.1 % (p = 0.027) and TMF decreased 17.6 % (p = 0.041). Disc degeneration (p \ 0.001) and its interaction with laminectomy (p \ 0.031) did significantly affect ETS. In the mildly degenerated group, ETS decreased 19.7 % from 7.6 Nm/degree (6.4-8.4) to 6.1 Nm/degree (1.5-10.3) following laminectomy. In the severely degenerated group, ETS decreased 22.3 % from 12.1 Nm/degree (4.6-21.9) to 9.4 Nm/degree (5.6-14.3) following laminectomy. In segments with low BMD, TMF was 40.7 % (p \ 0.001) lower than segments with high BMD [34.9 Nm (range 23.7-51.2) versus 58.9 Nm (range 43.8-79.2)]. Conclusions Laminectomy affects both torsion stiffness and torsion load to failure. In addition, torsional strength is strongly affected by BMD whereas disc degeneration affects torsional stiffness. Assessment of disc degeneration and BMD pre-operatively improves the understanding of the biomechanical effects of a lumbar laminectomy.

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a b s t r a c tIn loading experiments on the lumbar spine, typically three consecutive loading cycles are applied of which the third cycle is used for analysis. The aim of this study was to investigate whether the use of ten instead of three loading cycles reduces effects of viscoelastic behavior in the assessment of range of motion (ROM) and stiffness around the neutral orientation of the human lumbar spine. To this end, twelve cadaveric human lumbar spines (L1-L5) were obtained (mean age: 76.9 years). Before testing, the spines were subjected to a compressive load of 250 N for 1 h. To each spine, ten consecutive loading cycles were applied (À 4 Nm toþ4 Nm) in flexion and extension (FE), lateral bending (LB) and axial rotation (AR). The ROM and stiffness within the neutral zone were calculated per motion segment (L2-L3, L3-L4 or L4-L5) from load-displacement data. It was found that the ROM increased significantly (all p o 0.001) in all directions after three (FE: 0.07 degree/1.0%, LB: 0.08 degree/1.5%, and AR: 0.04 degree/1.5%) and after ten loading cycles (FE: 0.20 degree/2.9%, LB: 0.16 degree/3.3%, and AR: 0.09 degree/3.3%). Stiffness was not significantly affected, but varied considerably over cycles. Although effects were small, assessment of the tenth cycle instead of the third cycle reduces viscoelastic effects in repeated measurements of ROM, because the spine is closer to a steady state condition, while averaging over loading cycles would improve the assessment of stiffness estimates.

The loaded disk culture system is an intervertebral disk (IVD)-oriented bioreactor developed by the VU Medical Center (VUmc, Amsterdam, The Netherlands), which has the capacity of maintaining up to 12 IVDs in culture, for approximately 3 weeks after extraction. Using this system, eight goat IVDs were provided with the essential nutrients and submitted to compression tests without losing their biomechanical and physiological properties, for 22 days. Based on previous reports (Paul et al., 2012, 2013; Detiger et al., 2013), four of these IVDs were kept in physiological condition (control) and the other four were previously injected with chondroitinase ABC (CABC), in order to promote degenerative disk disease (DDD). The loading profile intercalated 16 h of activity loading with 8 h of loading recovery to express the standard circadian variations. The displacement behavior of these eight IVDs along the first 2 days of the experiment was numerically reproduced, using an IVD osmo-poro-hyper-viscoelastic and fiber-reinforced finite element (FE) model. The simulations were run on a custom FE solver (Castro et al., 2014). The analysis of the experimental results allowed concluding that the effect of the CABC injection was only significant in two of the four IVDs. The four control IVDs showed no signs of degeneration, as expected. In what concerns to the numerical simulations, the IVD FE model was able to reproduce the generic behavior of the two groups of goat IVDs (control and injected). However, some discrepancies were still noticed on the comparison between the injected IVDs and the numerical simulations, namely on the recovery periods. This may be justified by the complexity of the pathways for DDD, associated with the multiplicity of physiological responses to each direct or indirect stimulus. Nevertheless, one could conclude that ligaments, muscles, and IVD covering membranes could be added to the FE model, in order to improve its accuracy and properly describe the recovery periods.