Anterior cervical discectomy and fusion (ACDF) is a well-established spinal operation for cervical disc degeneration disease with neurological compromise. The procedure involves an anterior approach to the cervical spine with discectomy to relieve the pressure on the impinged spinal cord to slow disease progression. The prosthetic cage replaces the disc and can be inserted stand-alone or with an anterior plate that provides additional stability. The literature demonstrates that the cage-alone (CA) is given preference over the cage-plate (CP) technique due to better clinical outcomes, reduced operation time and resultant morbidity. This retrospective case-controlled study compared CA versus CP fixation used in single and multilevel anterior cervical discectomy and fusion for myelopathy in a tertiary centre in Wales. A retrospective clinico-radiological analysis was undertaken, following ACDF procedures over seven years in a single tertiary centre. Inclusion criteria were patients over 18 years of age with cervical myelopathy who had at least six-month follow-up data. SPSS was used to identify any statistically significant difference between both groups. The data were analysed to evaluate the consistency of our findings in comparison to published literature. Eighty-six patients formed the study cohort; 28 [33%] underwent ACDF with CA and 58 [67%] with CP. The patient demographics were similar in both groups, and fusion was observed in all individuals. There was no statistical difference between the two constructs when assessing subsidence, clinical complication (dysphagia, dysphonia, infection), radiological parameters and reoperations. However, a more significant percentage [43% v 61%] of patients improved their cervical lordosis angle with CP treatment. Furthermore, the study yielded that surgery to upper cervical levels results in a higher incidence of dysphagia [65% v 35%]. Finally, bony growth across the cage was observed on X-ray in 12[43%] patients, a unique finding not mentioned in the literature previously. Our study demonstrates no overall difference between the two groups, and we recommend careful consideration of individual patient factors when deciding what construct to choose

Degenerative disc disease (DDD) is a common cause of lower back pain. Calcification of the intervertebral disc (IVD) has been correlated with DDD, and is especially prevalent in scoliotic discs. The appearance of calcium deposits has been shown to increase with age, and its occurrence has been associated with several other disorders such as hyperparathyroidism, chondrocalcinosis, and arthritis. Trauma, vertebral fusion and infection have also been shown to increase the incidence of IVD calcification. Our data indicate that Ca. 2+. and expression of the extracellular calcium-sensing receptor (CaSR) are significantly increased in mild to severely degenerative human IVDs. In this study, we evaluated the effects of Ca. 2+. and CaSR on the degeneration and calcification of IVDs. Human donor lumbar spines of Thompson grade 2, 3 and 4 through organ donations within 24 hs after death. IVD cells, NP and AF, were isolated from tissue by sequential digestion with Pronase followed by Collagenase. Cells were expanded for 7 days under standard cell culture conditions. Immunohistochemistry was performed on IVD tissue to validate the grade and expression of CaSR. Free calcium levels were also measured and compared between grades. Immunocytochemistry, Western blotting and RT-qPCR were performed on cultured NP and AF cells to demonstrate expression of CaSR, matrix proteins aggrecan and collagen, catabolic enzymes and calcification markers. IVD cells were cultured in increasing concentrations of Ca. 2+. [1.0-5.0 mM], CaSR allosteric agonist (cincalcet, 1 uM), and IL-1b [5 ng/mL] for 7 days. Ex vivo IVD organ cultures were prepared using PrimeGrowth Disc Isolation System (Wisent Bioproducts, Montreal, Quebec). IVDs were cultured in 1.0, 2.5 mM Ca. 2+. or with cinacalcet for 21 days to determine effects on disc degeneration, calcification and biomechanics. Complex modulus and structural stiffness of disc tissues was determined using the MACH-1 mechanical testing system (Biomomentum, Laval, Quebec). Ca. 2+. dose-dependently decreased matrix protein synthesis of proteoglycan and Col II in NP and AF cells, similar to treatment with IL-1b. (n = 4). Contrarily to IL-1b, Ca. 2+. and cincalcet did not significantly increase the expression of catabolic enzymes save ADAMTS5. Similar effects were observed in whole organ cultures, as Ca. 2+. and cinacalcet decreased proteoglycan and collagen content. Although both Ca. 2+. and cinacalcet increased the expression of alkaline phosphatase (ALP), only in Ca. 2+. -treated IVDs was there evidence of calcium deposits in NP and AF tissues as determined by von Kossa staining. Biomechanical studies on Ca. 2+. and cinacalcet-treated IVDs demonstrated decreases in complex modulus (p<0.01 and p<0.001, respectively; n=5), however, only Ca. 2+. -treated IVDs was there significant increases stiffness in NP and AF tissues (p<0.001 and p<0.05, respectively; n=3). Our results suggest that changes in the local concentrations of calcium and activation of CaSR affects matrix protein synthesis, calcification and IVD biomechanics. Ca. 2+. may be a contributing factor in IVD degeneration and calcification

Testing potential therapeutics in the regeneration of the disc requires the use of model systems. Although several animal models have been developed to test intervertebral disc (IVD) regeneration, application becomes costly when used as a screening method. The bovine IVD organ culture system offers an inexpensive alternative, however, in the current paradigm, the bony vertebrae is removed to allow for nutrient diffusion to disc cells. This provides limitations on the conditions and strategies one can employ in investigating IVD regeneration and mechanisms in degenerative disc disease (i.e. complex loading). Although one method has been attempted to extend the survival of bovine vertebrae containing IVDs (vIVD) cell viability declined after two weeks in culture. Our goal was to develop and validate a long-term organ culture model with vertebral bone, which could be used subsequently for studying biological repair of disc degeneration and biomechanics. Preparation of vIVDs: Bovine IVDs from the tails of 22–28-month-old steers were prepared for organ culture by parallel cuts through the adjacent vertebral bodies at 1cm from the endplates using an IsoMet®1000 Buehler precision sectioning saw. vIVDs were split into two groups: IVDs treated with PrimeGrowth Media kit (developed by Intervertech and licensed to Wisent Bioproducts) and IVDs with DMEM. The PrimeGrowth group was incubated for 1h in PrimeGrowth Isolation Medium (Cat# 319–511-EL) and the DMEM group for 1h in DMEM. After isolation, IVDs were washed in PrimeGrowth Neutralisation Medium (Cat# 319–512-CL) while the other IVDs were washed in DMEM. The discs isolated with PrimeGrowth and DMEM were cultured for up to 5 months in sterile vented 60 ml Leakbuster™ Specimen Containers in PrimeGrowth Culture Medium (Cat# 319–510-CL) and DMEM with no mechanical load applied. Live/Dead Assay: vIVDs cultured for 1 or 5 months were dissected and cell viability was assessed in different regions by confocal microscopy using Live/Dead® (Invitrogen) fluorescence assay. Glucose Diffusion: After one month of culture, vIVDs were incubated for 72h in diffusion medium containing PBS (1x), CaCl2 (1mM), MgCl2 (0.5mM), KCl2 (5mM), 0.1% BSA and 150µM 2-NDBG, a D-glucose fluorescent analogue. Discs were dissected and IVD tissues were incubated in guanidinium chloride extraction buffer. Extracts were measured for fluorescence. After 5 months in culture, vIVDs prepared with PrimeGrowth kit demonstrated approximately 95% cell viability in all regions of the disc. However, dramatic reductions (∼90%) in vIVD viability were measured in DMEM group after 1 month. vIVD viability was related to the amount of 2-NDBG incorporated into the disc tissue. We have developed a novel method for isolating IVDs with vertebral bone capable of long-term viability. This method may not only help in the discovery of novel therapeutics in disc regeneration, but could also advance our understanding on complex loading paradigms in disc degeneration

Tungsten has been increasing in demand for use in manufacturing and recently, medical devices, as it imparts flexibility, strength, and conductance of metal alloys. Given the surge in tungsten use, our population may be subjected to elevated exposures. For instance, embolism coils made of tungsten have been shown to degrade in some patients. In a cohort of breast cancer patients who received tungsten-based shielding for intraoperative radiotherapy, urinary tungsten levels remained over tenfold higher 20 months post-surgery. In vivo models have demonstrated that tungsten exposure increases tumor metastasis and enhances the adipogenesis of bone marrow-derived mesenchymal stem cells while inhibiting osteogenesis. We recently determined that when mice are exposed to tungsten [15 ppm] in their drinking water, it bioaccumulates in the intervertebral disc tissue and vertebrae. This study was performed to determine the toxicity of tungsten on intervertebral disc. Bovine nucleus pulposus (bNP) and annulus fibrosus (bAF) cells were isolated from bovine caudal tails. Cells were expanded in flasks then prepared for 3D culturing in alginate beads at a density of 1×10. ∧. 6 cells/mL. Beads were cultured in medium supplemented with increasing tungsten concentrations in the form of sodium tungstate [0, 0.5, 5, 15 ug/mL] for 12 days. A modified GAG assay was performed on the beads to determine proteoglycan content and Western blotting for type II collagen (Col II) synthesis. Cell viability was determined by counting live and dead cells in the beads following incubation with the Live/Dead Viability Assay kit (Thermo Fisher Scientific). Cell numbers in beads at the end of the incubation period was determined using Quant-iT dsDNA Assay Kit (Thermo Fisher Scientific). Tungsten dose-dependently decreased the synthesis of proteoglycan in IVD cells, however, the effect was significant at the highest dose of 15 ug/mL. (n=3). Furthermore, although tungsten decreased the synthesis of Col II in IVD cells, it significantly increased the synthesis of Col I. Upregulation of catabolic enzymes ADAMTS4 and −5 were also observed in IVD cells treated with tungsten (n=3). Upon histological examination of spines from mice treated with tungsten [15 ug/mL] in their drinking water for 30 days, disc heights were diminished and Col I upregulation was observed (n=4). Cell viability was not markedly affected by tungsten in both bNP and bAF cells, but proliferation of bNP cells decreased at higher concentration. Surprisingly, histological examination of IVDs and gene expression analysis demonstrated upregulation of NGF expression in both NP and AF cells. In addition, endplate capillaries showed increases in CGRP and PGP9.5 expression as determined on histological sections of mouse IVDs, suggesting the development of sensory neuron invasion of the disc. We provide evidence that prolonged tungsten exposure can induce disc fibrosis and increase the expression of markers associated with pain. Tungsten toxicity may play a role in disc degeneration disease

Introduction. Anterior cervical decompression and fusion (ACDF) is considered a standard surgical treatment to degenerative discogenic diseases. Lately, the question arises whether or not ACDF significantly influences the progression of adjacent disc degeneration (ADD). The etiology of ADD is obscure and it has not been fully understood whether ADD is a consequence of fusion or it represents the aging pathway of the degenerative cervical process, thus making it a controversial topic [1-3]. There have been several discussions about the possibility of ACDF altering biomechanical conditions at adjacent segments, therefore resulting in increased load and excessive motion [3,4]. The purpose of this study was to compare the cervical segmental motion pre- and post-ACDF using novel 3D analytical techniques. Methods. Nine patients (2F/7M, mean age: 54.1 years, range 36–76 y.o.) underwent ACDF due to symptomatic cervical degenerative discogenic disease. One-level ACDF was performed in 4 patients, whereas 2-level ACDF was done in five, using cylindrical titanium porous cage implants. Pre- and post (postoperative periods ranged from 11-months, 25 days to 12-months, 22 days, mean postoperative period: 12.09 months) surgery, dynamic-CT examinations were conducted in neutral, flexion and extension positions. Subject-based 3D CT models were created for segmental motion analysis (Fig. 1). Six-degrees-of-freedom 3D segmental movements were analyzed using a validated Volume-Merge methods (accuracy: 0.1 mm in translation, 0.2°in rotation) [5]. The segmental translation was evaluated by the segmental translations of gravity centers of endplates (Fig. 2). Disc-height distribution was measured using a custom-written Visual C++ routine implementing a lease-distance calculation algorithm. The mean translation distance was calculated for the each adjacent level (Fig. 2). Differences of segmental motions and mean disc height between pre- and post-surgery at each level were compared by the Wilcoxon signed rank test. Results were presented mean±SEM. Results. Regarding the fusion level, the data shows decreases in both the flexion/extension (F/E) angular range of motion (ROM) (7.46±1.17°preoperatively vs. 3.14±0.56°post-operatively, p<0.003) and the segmental translation in the anterior/posterior direction (AP translation) after surgery (1.22±0.20 mm pre-operatively and 0.32±0.11 mm post-operatively, p<0.01). For the adjacent levels category (inferior and superior combined), the E/F angular ROM was larger after surgery (6.74±1.22°pre-operatively vs. 8.48±0.56°post-operatively, p<0.03). The lateral and axial rotational angular ranges of motion pre- and post-surgery did not show any statistically differences at the adjacent levels. The AP translation at the adjacent levels did not change after surgery (1.22±0.26 mm pre-operatively and 1.45±0.29 mm post-operatively). Translations in lateral and cranio-caudal directions also did not show change following surgery. The mean disc height in the adjacent level (2.39±0.14 mm) showed no differences with respect to the post-surgical measurements (2.40±0.19 mm). Conclusions. The use of a high-accuracy in vivo 3D kinematic analysis method enabled the detection of subtle changes in segmental movement between pre- and post-ACDF conditions. The result of the current study showed increased segmental movements in F/E angles at the adjacent level. These results are consistent with the some previous studies in the literature [4,6-11]. The magnitude of the increased movement, however, was only 1.74°from full-full-flexion to full-extension and no increase was found in AP translation. No disc height loss associated with disc degeneration was observed during a 1-year period after ACDF. Longer follow-up studies with larger patient cohorts will be required to investigate whether the increased F/E angle at the adjacent level effectively causes symptomatic ADD

Calcification of the intervertebral disc (IVD) has been correlated with degenerative disc disease (DDD), a common cause of low back pain. The appearance of calcium deposits has been shown to increase with age, and its occurrence has been associated with several other disorders such as hyperparathyroidism, chondrocalcinosis, and arthritis. Trauma, vertebral fusion and infection have also been shown to increase the incidence of IVD calcification. The role of IVD calcification in the development DDD is unknown. Our preliminary data suggest that ionic calcium content and expression of the extracellular calcium-sensing receptor (CaSR), a G protein-coupled receptor (GPCR) and regulator of calcium homeostasis, are increased in the degenerated discs. However, its role in DDD remains unclear. IVD Cells: Bovine and normal human IVD cells were incubated in PrimeGrowth culture medium (Wisent Bioproducts, Canada; Cat# 319–510-CL, −S1, and S2) and supplemented with various concentrations of calcium (1.0, 1.5, 2.5, 5.0 mM), a CaSR agonist [5 µM], or IL-1β [10 ng/ml] for 7 days. Accumulated matrix protein was quantitated for aggrecan and type II collagen (Col II) by Western blotting. Conditioned medium was also collected from cells treated for 24h and measured for the synthesis and release of total proteoglycan using the DMMB assay and Western blotting for Col II content. IVD Cultures: Caudal IVDs from tails of 20–24 month old steers were isolated with the PrimeGrowth Isolation kit (Wisent Bioproducts, Canada). IVDs were cultured for 4 weeks in PrimeGrowth culture medium supplemented with calcium (1.0, 2.5, or 5.0 mM), or a CaSR agonist [5 µM]. Cell viability was measured in NP and AF tissue using Live/Dead Imaging kit (ThermoFisher, Waltham, MA), to determine if Ca2+ effects cell viability end the expression of aggrecan and Col II was evaluated in the IVD tissue by Western blotting. Histological sections were prepared to determine total proteoglycan content, alkaline phosphatase expression and degree of mineralisation by von Kossa staining. The accumulation of aggrecan and Col II decreased dose-dependently in IVD cells following supplementation with calcium or the CaSR agonist. Conditioned medium also demonstrated decreases in the synthesis and release of proteoglycan and collagen with increasing Ca2+ dose or direct activation of the CaSR with agonist. A similar phenomenon was observed for total proteoglycan and aggrecan and Col II in IVDs following calcium supplementation or the CaSR agonist. In addition to decreases in Col II and aggrecan, increases in alkaline phosphatase expression and mineralisation was observed in IVDs cultured in elevated Ca2+ concentrations without affecting cell viability. Our results suggest that changes in the local concentrations of calcium are not benign, and that activation of the CaSR may be a contributing factor in IVD degeneration. Determining ways to minimise Ca2+ infiltration into the disc may mitigate disc degeneration

Stable thoracolumbar fracture is a common injury. The factors that determine its outcome are unclear. Aspects of injury severity were analysed for their ability to predict outcome by controlling other outcome-affecting factors (patient's pre-injury health status, legal aspects, associated injuries, etc.). No reliable disc injury severity grading system was available and therefore a new system was developed. A prospective observational study of 44 conservatively treated patients with stable fractures between T11 and L5 was conducted. Bony injury severity was scored based on comminution, apposition and kyphosis parameters. Disc injury severity was scored by the new scale based on variables – Herniation, Indentation, Height decrease and Signal change – seen in MRI. Ten outcome domains (five domains of pain and function each) were assessed at 1 to 2 years from injury. The data was analysed by non-parametric correlation and stepwise-linear regression analysis to assess the predictive value of different variables (patient factors, injury factors and social factor) to outcome. The correlation coefficients between injury severity and outcome were consistently higher with disc injury severity than bony. Disc injury severity showed highest predictive value for both pain (29%) and functional (16%) outcomes, whereas the bony injury severity parameters (kyphosis, etc.) and the posterior ligament injury severity provided no prediction of outcome. According to AO classification, the fractures were A1, A2, A3 and B1; in this spectrum of injuries, the AO classification had no prediction of outcome. The disc injury score also had a good predictive value for final disc degeneration. Disc injury severity should be gauged in advising prognosis and treatment. The new disc injury severity grading system showed good construct validity

Purpose. Disc degeneration is known to occur early in adult life, but at present there is no medical treatment to reverse or even retard the problem. Development of medical treatments is complicated by the lack of a validated long term organ culture model in which therapeutic candidates can be studied. The objective of this study was to optimize and validate an organ culture system for intact human intervertebral disc (IVD), which could be used subsequently to determine whether synthetic peptide growth factors can stimulate disc cell metabolism and initiate a repair response. Method. Seventy lumbar IVDs, from 14 individuals, were isolated within 24 h after death. Discs were prepared for organ culture by removing bony endplates but retaining cartilaginous endplates (CEP). Discs were cultured with no external load applied. The effects of glucose and FBS concentrations were evaluated. Dulbeccos Modified Eagle Media (DMEM) was supplemented with glucose, 4.5g/L or 1g/L, referred to as high and low (physiological) glucose, and FBS, 5% or 1%, referred to as high and low FBS, respectively. After a four week culture period, samples were taken across the disc using a 4 mm biopsy punch. Cell viability was analyzed using a live/dead fluorescence assay (Live/Dead, Invitrogen) and visualized by confocal microscopy. CEP discs were also placed in long term culture for four months, and cell viability was assessed. Western bolt analysis for the G1 domain of aggrecan was also performed to assess the effect of nutritional state on disc catabolism. Results. Cell viability in CEP isolated discs was evaluated after four weeks and four months of organ culture under high and physiological nutritional state. Previous studies have shown that high glucose levels are needed to maintain cell viability in organ culture, but in our model 96–98% live cells were present throughout the disc independent of FBS and glucose levels and the duration of culture tested. Western blot probing for the G1 domain of aggrecan showed no difference with the change of nutritional state across all regions indicating that low nutritional state had no detrimental effect on disc metabolism. Conclusion. We have developed a novel technique for isolation and culturing of intact IVDs. The described CEP system maintained sufficient nutrient supply and high cell survival in all regions of the disc for up to four months of culture also under physiological culturing condition. As the CEP system maintains high cell viability in long term cultures, it is a suitable model in which the regenerative effect of various bioactive peptides can be studied. The availability of an intact disc organ culture system has considerable advantage over the culture of isolated disc cells, as it maintains the cells in their unique microenvironment, so making any response to catabolic or anabolic agents more physiologically relevant

Purpose. Disc degeneration is known to occur early in adult life, but at present there is no medical treatment to reverse or even retard the problem. Development of medical treatments is complicated by the lack of a validated long term organ culture model in which therapeutic candidates can be studied. The objective of this study was to optimize and validate an organ culture system for intact human intervertebral disc (IVD), which could be used subsequently to determine whether synthetic peptide growth factors can stimulate disc cell metabolism and initiate a repair response. Method. Seventy lumbar IVDs, from 14 individuals, were isolated within 24 h after death. Discs were prepared for organ culture by removing bony endplates but retaining cartilaginous endplates (CEP). Discs were cultured with no external load applied. The effects of glucose and FBS concentrations were evaluated. Dulbeccos Modified Eagle Media (DMEM) was supplemented with glucose, 4.5g/L or 1g/L, referred to as high and low (physiological) glucose, and FBS, 5% or 1%, referred to as high and low FBS, respectively. After a four week culture period, samples were taken across the disc using a 4 mm biopsy punch. Cell viability was analyzed using a live/dead fluorescence assay (Live/Dead, Invitrogen) and visualized by confocal microscopy. CEP discs were also placed in long term culture for four months, and cell viability was assessed. Western bolt analysis for the G1 domain of aggrecan was also performed to assess the effect of nutritional state on disc catabolism. Results. Cell viability in CEP isolated discs was evaluated after four weeks and four months of organ culture under high and physiological nutritional state. Previous studies have shown that high glucose levels are needed to maintain cell viability in organ culture, but in our model 96–98% live cells were present throughout the disc independent of FBS and glucose levels and the duration of culture tested. Western blot probing for the G1 domain of aggrecan showed no difference with the change of nutritional state across all regions indicating that low nutritional state had no detrimental effect on disc metabolism. Conclusion. We have developed a novel technique for isolation and culturing of intact IVDs. The described CEP system maintained sufficient nutrient supply and high cell survival in all regions of the disc for up to four months of culture also under physiological culturing condition. As the CEP system maintains high cell viability in long term cultures, it is a suitable model in which the regenerative effect of various bioactive peptides can be studied. The availability of an intact disc organ culture system has considerable advantage over the culture of isolated disc cells, as it maintains the cells in their unique microenvironment, so making any response to catabolic or anabolic agents more physiologically relevant

The emerging of non-fusion surgery is aimed to solve the long-term complication of fusion surgery that may bring the adjacent disc degeneration. Among several kinds of artificial discs developed in these years, the majority in the market is Prodisc-L (Synthes Inc.) which is designed with the purpose to restore the motions including anteroposterior translation, lateral bending, and axial rotation. These is also one artificial disc called Physio-L (Nexgen Spine) which were hyper-elastic material (Polycarbonate Polyurethanes) and is designed to restore the motions maintioned above plus axial loading. The concept of using hyper-elastic material as disc is to mimic the material properties of intervetebral discs so that this disc both absorb the axial loading and also restore the physiological range of motion. Few studies focused on the biomechanical behavior of hyper-elastic artificial discs have yet been reported. Therefore, the purpose of this study is to compare the biomechanical behavior between Prodisc-L and Physio-L. A validated three-dimensional finite element model of the L1-L5 lumbar intact spine was used in this study with ANSYS software [Fig.1]. Total disc replacement surgery, partial discectomy, total nuclectomy and removal of the anterior longitudinal ligament were performed at the L3/L4 segment of this intact model, and the Prodisc-L and Physio-L was implanted into L3/L4 segment, respectively. In addition, hyper-elastic materials adopted by Physio-L are usually categorized by their hardness into soft and hard [Fig.2]. Therefore, two kinds of Physio-L were studied. A 400 N follower load and a 10 N-m moment were applied to the intact model to obtain four physiological motions as comparison baseline. The implanted models were subjected to 400 N follower load and specific moments in accordance with the hybrid test method. For the Prodisc-L model in the surgical segment, the range of motion (ROM) varied by -26%, +17%, -0.01%, and -0.04% in flexion, extension, lateral bending, and axial rotation, respectively, as compared to intact model [Fig.3]. For the Physio-L (soft) model, ROM varied by +10%, +8%, +3%, and +19% in four physiological motions, respectively. For the physio-L (hard) model, ROM varied by +1%, +8%, +1%, and +11% in four physiological motions, respectively. For the Prodisc-L model in the adjacent segments, ROM varied by +4% ∼ +10%, -2% ∼ -5%, -1% ∼ -4%, and +1% ∼ -2% in four physiological motions, respectively. For the Physio-L (soft) model, ROM varied by 0% ∼ -5%, -2% ∼ -5%, -0% ∼ -5%, and -9% ∼ -11% in four physiological motions, respectively. For the physio-L (hard) model, ROM varied by +4% ∼ -2%, +8% ∼ -5%, +1 ∼ -5%, and +11% ∼ -6% in four physiological motions, respectively. As seemed in the simulation, the behavior of Physio-L (both soft and hard) is similar to that of intact model under flexion and extension, but not in axial rotation. In addition, Physio-L (hard) model is more similar to intact model as compared to Physio-L (soft) model