Introduction. Vertebral compression fractures are the most common type of osteoporotic fracture. Though 89% of clinical fractures occur anteriorly, it is challenging to replicate these ex vivo with the underlying intervertebral discs (IVDs) present. Furthermore, the role of
Purpose of study and background. Degeneration of the intervertebral disc is a strong contributor of low back pain. Studies have shown that both, mechanical unloading and overloading, lead to
Intervertebral disc degeneration (DD) is a complex age-related condition that constitutes the main risk factor for disabling back pain. DD is assessed using different traits extracted from MR imaging (MRI), normally combined to give summary measures (e.g. Pfirmann score). The aetiology of DD is poorly understood and despite its high heritability (75%), the precise genetic predisposition is yet to be defined. Genome wide association study (GWAS) is used to discover genetic variants associated with a disease or phenotype. It tests variants across the whole genome. It requires large samples to provide adequate but unfortunately there is poor availability of spine imaging data due to the high cost of MRI. We have adopted new methods to examine different MRI traits independently and use the information of those traits to boost GWAS power using specialized statistical software for jointly analyse correlated traits. We examined DD MRI features disc narrowing, disc bulge, disc signal intensity and osteophyte formation in the TwinsUK cohort who had undergone T2-weighted sagittal spine MRI. GWAS were performed on the four traits. MTAG software was used to boost single trait GWAS power using the information in the other trait GWAS. 9 different loci were identified.Background
Methods/Results
Introduction. Risk factors for
Introduction. Delamination of the annulus fibrosus is an early feature of
Summary Statement. Spinal flexibility in bending and axial torque has been shown to exhibit very modest changes with advancing
Purpose &
Relevance: To examine the prevalence of degenerative findings in the thoracic spine in a population sample of adult men. Normative data on thoracic degenerative findings provide an essential reference for related observations in patients. Methods &
Results: Qualitative and quantitative assessments of thoracic
This short contribution aims to explain how intervertebral disc ‘degeneration’ differs from normal ageing, and to suggest how mechanical loading and constitutional factors interact to cause
Mechanical function and failure of intervertebral discs. In a healthy disc, the nucleus pulposus acts like a pressurised fluid which is restrained by tensile stress within the annulus. With increasing age, the nucleus becomes more fibrous, and biochemical changes cause the whole disc to become less elastic, and more yellow in colour. Mechanically, the hydrostatic nucleus shrinks with age, and concentrations of compressive stress appear in the posterior annulus. Experiments on cadaveric spines have shown that healthy discs can prolapse when loaded severely or repetitively in bending and compression, and that internal disruption of the disc probably follows damage to the vertebral endplates. However, mechanical loading is not necessarily harmful to living discs: on the contrary, moderate repetitive loading may lead to disc hypertrophy rather than injury.
Introduction:
Purpose: To evaluate the long term clinical outcomes as well as radiological changes in distal unfused mobile segments and to evaluate factors that may predispose to distal
Osteoporotic fractures are associated with bone loss following hormonal changes and reduced physical activity in middle age. But these systemic changes do not explain why the anterior vertebral body should be such a common site of fracture. We hypothesise that age-related degenerative changes in the intervertebral discs can lead to abnormal load-bearing by the anterior vertebral body. Cadaveric lumbar motion segments (mean age 50 ± 19 yrs, n = 33) were subjected to 2 kN of compressive loading while the distribution of compressive stress was measured along the antero-posterior diameter of the intervertebral disc, using a miniature pressure-transducer. “Stress profiles” were obtained with each motion segment positioned to simulate a) the erect standing posture, and b) a forward stooping posture. Stress measurements were effectively integrated over area in order to calculate the force acting on the anterior and posterior halves of the disc (
. 1. ). These two forces were subtracted from the applied 2 kN to determine the compressive force resisted by the neural arch. Discs were sectioned and their degree of
Introduction. Primary cilia are singular structures containing a microtubule-based axoneme which are believed to not only be mechanosensitive but also to co-ordinate many cell functions via signalling pathways including Hedgehog and Wnt. Primary cilia have previously been described on cells of mouse intervertebral discs (IVDs), but not in bovine or human IVDs. Our aim was to examine primary cilia in these species. Methods. Nucleus pulposus cells were obtained from cows with no overt
Summary Statement. DNA damage induced by systemic drugs or local γ-irradiation drives
Introduction Disc replacement surgery is being investigated as an alternative to spinal fusion surgery in the hope that maintaining segment spinal motion will not only relieve pain, but also prevent or reduce the likelihood of symptomatic adjacent segment degeneration that is believed to be a consequence of fusion surgery. The aim of this study was to identify evidence in the medical literature that indicates whether or not spinal fusion surgery increases the likelihood of symptomatic adjacent segment degeneration compared to disc replacement surgery or natural history. Methods A search of the Cochrane Controlled Trials Register, Medline and reference lists of retrieved articles was performed. Search terms included arthroplasty replacement, spinal fusion, prognosis, controlled clinical trials and cohort studies, Studies were included if abstracts were available electronically, were published in the English language before1/3/2005 and involved humans. Levels of evidence were determined using the Oxford Centre for Evidence-Based Medicine criteria (. http://www.cebm.net/levels of evidence.asp. ). Discussion The majority of identified studies were case series of patients presenting with adjacent level
Study Design: Experimental in vivo study on New Zealand White Rabbits. Summary of Background Data: We have developed an in-vivo rabbit model of lumbar
Objectives. A single degenerate intervertebral disc is suggested to promote rapid degeneration in its adjacent discs. We validated this hypothesis using discordant co-twin case-control design. Methods. 185 pairs of twins were selected from the TwinsUK database having cervical MRI scans at baseline and at follow-up, after 10 years. Isolated
Introduction and Aims: Lower back and/or leg pain is a symptom of a number of pathological conditions involving lumbosacral nerve roots. Disc herniation is one of the most common causes of LBP (after mechanical back pain). There is controversy regarding the progression of
Introduction: The development of laboratory techniques in the last ten years has enabled the successful harvest, in vitro selection, culture and transplant of chondrocytes. The study proposes that transplantation of autologous chondrocytes prevents degeneration of the intervertebral disc following outer annular injury in an ovine model. Methods: Eight sheep were anaesthetised and five contiguous lumbar discs were exposed via a left-sided posterolateral approach. Four of the animals were given full thickness annular incisions in three alternate discs. No annular incisions were made in the other 4 sheep. Costal cartilage was harvested from the left twelfth rib of all animals. Tissue was cultured and the chondrocytes were labelled in vitro with CFSE for verification following transplantation. Six weeks later autologous cultured chondrocytes were injected into the lower two alternate discs of all animals, leaving the uppermost discs and those untouched in between as internal controls. Animals were sacrificed after three, six, twelve and twenty-four weeks. Results were based on X rays, histological, and immunocytochemical assessments. Results: Preliminary histological results up to three months showed viability of cultured chondrocytes and matrix production post transplantation. Serial X rays suggested that progressive
INTRODUCTION: The development of laboratory techniques in the last ten years has enabled the successful harvest, in vitro selection, culture and transplant of chondrocytes. The study proposes that transplantation of autologous chondrocytes prevents degeneration of the intervertebral disc following outer annular injury in an ovine model. METHODS: Eight sheep were anaesthetised and five contiguous lumbar discs were exposed via a left-sided posterolateral approach. Four of the animals were given full thickness annular incisions in three alternate discs. No annular incisions were made in the other four sheep. Costal cartilage was harvested from the left twelfth rib of all animals. Tissue was cultured and the chondrocytes were labelled in vitro with CFSE for verification following transplantation. Six weeks later autologous cultured chondrocytes were injected into the lower two alternate discs of all animals, leaving the uppermost discs and those untouched in between as internal controls. Animals were sacrificed after three, six, twelve and twenty-four weeks. Results were based on X-rays, histological, and immunocytochemical assessments. RESULTS: Preliminary histological results up to three months showed viability of cultured chondrocytes and matrix production post transplantation. Serial X-rays suggested that progressive
Study Design and Objectives: The aim of this study is to analyse clinical results of total lumbar disc arthroplasty according to the type of
The lumbar spines of 22 patients were examined for
Introduction: The evidence of genetic background as an important causative factor in
Background. The majority of adults will experience an episode of low back pain during their life. Patients with non-specific low back pain and lumbar
Introduction The annulus fibrosus of the intervertebral disc is composed of a series of concentric collagenous lamellae that constrain the highly pressurised fluid of the nucleus pulposus. With advancing age and even after physical injury in youth the disc almost invariably becomes progressively degenerate due to the combined effects of dehydration of the nucleus and disruption of the annulus. There is conjecture however, about which of the two compartments shows degenerate changes first. Methods A histological and biochemical review is presented, based on a review of the literature and work carried out in our laboratories. Results Three distinct types of annulus tears are seen histologically. Rim tears are formed by detachment of the peripheral annular fibres from the vertebral rim. Autopsy studies show that these lesions are rare in subjects younger than 30 years but the incidence increases significantly with advancing age. Although granulation tissue grows into the outer layers of the annulus in a normal healing response, these lesions frequently extend deeper into the disc. Radiating tears course radially across several lamellae, most often extending from the vertebral rim across the nucleus to involve the posterior side of the disc. These lesions are seen mostly between the ages of 30 and 50 years. Concentric tears are characterised by separation of adjacent lamellae and may appear as early as the second decade of life. In advanced degeneration nuclear changes may be seen with any combination of annular lesions. Biochemical changes, including dehydration and reduced proteoglycan content of the nucleus, parallel the morphologic changes to the disc. Altered biochemistry is further reflected by MRI studies in which some scanning sequences can detect even modest loss of fluid from the nucleus as early as three months after experimental annular incision. Annular lesions also compromise the biomechanical properties of the disc. While internal fixation that aims to immobilise the injured disc may promote some recovery of the mechanical integrity, degeneration nevertheless advances in the long term. Attempts to seal peripheral annular defects in experimental studies using biocompatible glue have also failed to promote healing and to prevent progression of even minor structural defects. Conclusions Technological developments such as gene transfer into disc cells and direct implantation of either stem cells or more mature cells are emerging as potential candidates for the treatment of
Introduction. The usefulness of markers of non-specific low back pain (NSLBP), including MRI derived measurements of cross-sectional area (CSA) and functional CSA (FCSA, fat free muscle area) of the lumbar musculature, is in doubt. To our knowledge, such markers remain unexplored in Lumbar
Thirty-three patients who had undergone anterior cervical fusion for degenerative disc disease were reviewed to determine the efficacy of the procedure. Only patients who were available for examination and who had undergone operation at least one year previously were included in the review. Nearly all had had arm pain and three-quarters neck pain. Diminished neck movement and neurological abnormalities in the arms had been frequent findings. Diagnosis from the clinical features and plain radiographs is described. Myelography was not used routinely and discography was not used at all. Indications for operation and surgical technique are described. Results show that pain in the neck and arm was relieved in a high proportion of cases and that the neurological abnormalities often recovered. It is concluded that this operation is safe and has a definite place in the relief of pain from cervical
Study Design: Experimental study to assess tissue engineered solutions to
Background: The healthy, adult human disc is innervated but the nerves are restricted to the outer few millimetres of the annulus fibrosus. In degenerate discs with associated back pain, however, the nerves are more numerous and penetrate further in. We have used a sheep model of intervertebral disc degeneration to monitor the presence and organisation of nerves in the
The pathophysiology of intervertebral disc degeneration has been extensively studied. Various factors have been suggested as influencing its aetiology, including mechanical factors, such as compressive loading, shear stress and vibration, as well as ageing, genetic, systemic and toxic factors, which can lead to degeneration of the disc through biochemical reactions. How are these factors linked? What is their individual importance? There is no clear evidence indicating whether ageing in the presence of repetitive injury or repetitive injury in the absence of ageing plays a greater role in the degenerative process. Mechanical factors can trigger biochemical reactions which, in turn, may promote the normal biological changes of ageing, which can also be accelerated by genetic factors. Degradation of the molecular structure of the disc during ageing renders it more susceptible to superimposed mechanical injuries. This review supports the theory that degeneration of the disc has a complex multifactorial aetiology. Which factors initiate the events in the degenerative cascade is a question that remains unanswered, but most evidence points to an age-related process influenced primarily by mechanical and genetic factors.
We reviewed the radiographs of 325 unselected patients with defects in the pars interarticularis of L5 to study whether the incidence of vertebral slip in spondylolysis of L5 remained unchanged after the age of 20 years. MRI was also carried out on 111 of the patients to investigate the relationship between the shape of the transverse process of L5 and the degeneration of the discs adjacent to this level. The incidence of spondylolisthesis increased with age from 17% in the second decade to 51% in the sixth. The transverse process was significantly more slender in patients with less degeneration at L4/5 and advanced degeneration at L5/S1 than in patients with advanced degeneration at L4/5 and less degeneration at L5/S1. Vertebral slip secondary to an isthmic defect of L5 after the age of 20 years was confirmed and the adjacent
Introduction. Patients with recurrent low back pain (LBP) exhibit changes in postural control. Stereotypical muscle activations resulting from external perturbations include anticipatory (APAs) and compensatory (CPAs) postural adjustments. This study aimed to determine differences in postural control strategies (APAs and CPAs) between those with and without lumbar
Introduction: A computer model of the L4/5 human intervertebral disc is currently under development. An integral aspect of this model is the material properties assigned to its components. Detailed data on the material properties of the anulus fibrosus ground matrix are not available in the existing literature. To determine these properties, mechanical tests were carried out on specimens of anulus fibrosus harvested from sheep spines. The tests included unconfined uniaxial compression, simple shear and biaxial compression. Data on the strain required to cause permanent damage in the anulus ground matrix and data on the mechanical response of the anulus to repeated loading were obtained. Methods: Intervertebral discs were isolated from the lumbar spines of recently sacrificed sheep. These discs were sectioned into test specimens ensuring there were no continuous collagen fibres bearing load. The edge dimensions of the cubic specimens were 3 ± 0.2mm. To ascertain the strain to initiate tissue damage, the specimens underwent successive loadings, which were carried out 1 hour apart to allow recovery. The maximum strain in each test was increased incrementally by 5% until a reduction in stiffness was observed in the following test. Separate tests were carried out to quantify and characterise the response of the anulus ground matrix in the three modes of loading and to strains greater than that which initiates damage. Results: The strains at which permanent tissue damage occurred were between 20 and 27% in uniaxial compression and between 25 and 35% in simple shear. Testing the specimen beyond these strains showed an obvious reduction in stiffness. The biaxial compression tests showed similar changes but did not result in such pronounced losses in stiffness. The material characteristics were reproducible up to 20% strain. Following deformation to higher strains the altered mechanics were also shown to be reproducible indicating that the matrix had been deranged but not failed. Discussion: Average physiological strains in the L4/5 intervertebral disc are in the order of 10–50% based on maximum deformations observed in vivo. The current results demonstrate that this strain will cause some permanent damage to the anulus ground matrix, however, the matrix will still be capable of carrying stress upon repeated loading. Thompson et. al. 1. found that people over the age of 35 all exhibited signs of
INTRODUCTION: A computer model of the L4/5 human intervertebral disc is currently under development. An integral aspect of this model is the material properties assigned to its components. Detailed data on the material properties of the anulus fibrosus ground matrix are not available in the existing literature. To determine these properties, mechanical tests were carried out on specimens of anulus fibrosus harvested from sheep spines. The tests included unconfined uniaxial compression, simple shear and biaxial compression. Data on the strain required to cause permanent damage in the anulus ground matrix and data on the mechanical response of the anulus to repeated loading were obtained. METHODS: Intervertebral discs were isolated from the lumbar spines of recently sacrificed sheep. These discs were sectioned into test specimens ensuring there were no continuous collagen fibres bearing load. The edge dimensions of the cubic specimens were 3 ± 0.2 mm. To ascertain the strain to initiate tissue damage, the specimens underwent successive loadings, which were carried out one hour apart to allow recovery. The maximum strain in each test was increased incrementally by 5% until a reduction in stiffness was observed in the following test. Separate tests were carried out to quantify and characterise the response of the anulus ground matrix in the three modes of loading and to strains greater than that which initiates damage. RESULTS: The strains at which permanent tissue damage occurred were between 20 and 27% in uniaxial compression and between 25 and 35% in simple shear. Testing the specimen beyond these strains showed an obvious reduction in stiffness. The biaxial compression tests showed similar changes but did not result in such pronounced losses in stiffness. The material characteristics were reproducible up to 20% strain. Following deformation to higher strains the altered mechanics were also shown to be reproducible indicating that the matrix had been deranged but not failed. DISCUSSION: Average physiological strains in the L4/5 intervertebral disc are in the order of 10–50% based on maximum deformations observed in vivo. The current results demonstrate that this strain will cause some permanent damage to the anulus ground matrix, however, the matrix will still be capable of carrying stress upon repeated loading. Thompson et. al. 1. found that people over the age of 35 all exhibited signs of
Introduction: There are few long-term studies after Zielke ventral derotation spondylodesis (VDS). We present a minimum 17 year follow-up study to assess factors predicting distal adjacent
Low back pain is thought to relate to intervertebral disc (IVD) degeneration. Although the mechanisms have not been clearly identified, exhaustion of nucleus pulposus cells and their producing matrix is regarded as one cause. The matrix of the IVD is continuously replenished and remodeled by tissue-specialized cells and are crucial in supporting the IVD function. However, due to aging, trauma, and genetic and lifestyle factors, the cells can lose their potency and viability, thereby limiting their collective matrix production capacity. We have discovered the link between loss of angiopoietin-1 receptor (Tie2)-positive human NP progenitor cells (NPPC) and IVD degeneration. Tie2+ cells were characterized as undifferentiated cells with multipotency and possessing high self-renewal abilities. Thus we and others have proposed Tie2+ NPPC as a potent cell source for regenerative cell therapies against IVD degeneration. However, their utilization is hindered by low Tie2-expressing cell yields from NP tissue, in particular from commonly available older and degenerated tissue sources. Moreover, NPPC show a rapid Tie2 decrease due to cell differentiation as part of standard culture processes. As such, a need exists to optimize or develop new culture methods that enable the maintenance of Tie2-expressing NPPC. Trials to overcome these difficulties will be shared.
Background: Over several decades, investigators have been trying to identify the painful degenerate disc. Their work included two main methods. The first was to set criteria on the radiological investigations, mainly the MRI scan, to describe the severity of the degenerative disc disease (DDD); and the second was to perform discographies. Neither of these two methods precluded the need for the other. Purpose: Using Pfirrmann’s classification, we correlated static MRI images, for the severity of segmental
The factors influencing normal spine curvature in midlife are unknown. We performed an MR and plain radiograph study on well characterised, unselected twin volunteers from the TwinsUK register ( T2 weighted MR scans and long spine standing radiographs were obtained at the same morning visit on twin pairs. Midline sagittal MR images were coded for 4 degenerative features. SpineviewTM software was applied plain films and calculated the angles of curvature. A classical twin study was performed. Multivariate regression analysis was used to determine the association between spine curves, LDD and confounders (age, body mass index).Background
Methods
Aims. The presence of facet tropism has been correlated with an elevated susceptibility to lumbar disc pathology. Our objective was to evaluate the impact of facet tropism on chronic lumbosacral discogenic pain through the analysis of clinical data and finite element modelling (FEM). Methods. Retrospective analysis was conducted on clinical data, with a specific focus on the spinal units displaying facet tropism, utilizing FEM analysis for motion simulation. We studied 318 intervertebral levels in 156 patients who had undergone provocation discography. Significant predictors of clinical findings were identified by univariate and multivariate analyses. Loading conditions were applied in FEM simulations to mimic biomechanical effects on intervertebral discs, focusing on maximal displacement and intradiscal pressures, gauged through alterations in disc morphology and physical stress. Results. A total of 144 discs were categorized as ‘positive’ and 174 discs as ‘negative’ by the results of provocation discography. The presence of defined facet tropism (OR 3.451, 95% CI 1.944 to 6.126) and higher Adams classification (OR 2.172, 95% CI 1.523 to 3.097) were important predictive parameters for discography-‘positive’ discs. FEM simulations showcased uneven stress distribution and significant disc displacement in tropism-affected discs, where loading exacerbated stress on facets with greater angles. During varied positions, notably increased stress and displacement were observed in discs with tropism compared to those with normal facet structure. Conclusion. Our findings indicate that facet tropism can contribute to disc herniation and changes in intradiscal pressure, potentially exacerbating
The complex relationship between acetabular component position and spinopelvic mobility in patients following total hip arthroplasty (THA) renders it difficult to optimize acetabular component positioning. Mobility of the normal lumbar spine during postural changes results in alterations in pelvic tilt (PT) to maintain the sagittal balance in each posture and, as a consequence, markedly changes the functional component anteversion (FCA). This study aimed to investigate the in vivo association of lumbar degenerative disc disease (DDD) with the PT angle and with FCA during postural changes in THA patients. A total of 50 patients with unilateral THA underwent CT imaging for radiological evaluation of presence and severity of lumbar DDD. In all, 18 patients with lumbar DDD were compared to 32 patients without lumbar DDD. In vivo PT and FCA, and the magnitudes of changes (ΔPT; ΔFCA) during supine, standing, swing-phase, and stance-phase positions were measured using a validated dual fluoroscopic imaging system.Aims
Methods
One hundred and thirty-nine discs from cadaveric lumbar spines were injected with a mixture of radio-opaque fluid and dye. Discograms were taken and the discs were then sectioned in the sagittal plane. Examination of the sections revealed that injected fluid did not at first mix with the disc matrix but pushed it aside to form pools of injected fluid. The location of these pools, and hence the appearance of a discogram, depended on the stage of degeneration of the disc. It is concluded that useful clinical information can be obtained from discograms.
There are many difficulties associated with the localisation of the symptomatic segment in patients presenting with cervicobrachial pain with no evidence of impaired conduction in the nerve root. Ancillary radiological investigations such as myelography, epidural phlebography, and epidural myelograms are of unreliable diagnostic value. However, discography can be of value if the technique described here is used. Infiltration of the cervical nerve root with local anaesthetic has also proved useful in the localisation of the symptomatic segment. The techniques used in cervical discography and infiltration of the nerve root are described and their reliability is assessed.
CaSO4 and CaCO3 3% alginate hydrogels were injected into the NP cavity of a bovine tail. After 90 minutes the tail was dissected to reveal the gel. NP cells released from pooled bovine NP tissue were dispersed into the CaSO4 and CaCO3 alginate gels (10x106 cells.mL-1) with and without hylan (Synvisc®) and cultured for 21 days.
Injectable alginate suspensions formed solid viscoelastic gels, filling the exact shape of the NP cavity. NP DNA and ECM synthesis was significantly greater in the CaCO3 alginate gel than in the CaSO4 alginate gel (p<
0.05). Synvisc® significantly increased sulphated GAG (p<
0.01) and collagen (p<
0.05) production. These effects were supported histologically and immunohistologically where cells in the CaCO3 and Synvisc® gels stained more intensely for proteoglycan and collagen type II.
We studied 135 lumbar discs from 27 spines removed post-mortem from subjects of an average age of 31.5 years. Defects of the annulus fibrosus were classified as peripheral, circumferential or radiating; the nucleus pulposus as normal, moderately or severely degenerate. Peripheral tears were more frequent in the anterior annulus, except in the L5-S1 disc. Circumferential tears were equally distributed between the anterior and the posterior annulus. Almost all the radiating tears were in the posterior annulus, and closely related to the presence of severe nuclear degeneration. Histology suggested that peripheral tears were due to trauma rather than biochemical degradation, and that they developed independently of nuclear degeneration. The association of peripheral annular lesions with low back pain is uncertain but our study suggests that they may have a role in the pathogenesis of discogenic pain.
the mid-sagittal plane of 10 male and 10 female discs aged 13–79 years; 7 parasagittal slices through a single disc; discs showing various types of tears.
Most chondrocytes were unicellular but bicellular and multicellular chondrons were common in the margins of large tears and the nucleus in degenerate discs. Cellularity was highest in the right posterior quadrant, lowest in the left anterior quadrant, about equal in the left posterior and right anterior quadrants, and substantially higher in the right half of the nucleus. The correlation of increasing age with declining cell density was much stronger for the nucleus than for the annulus. Nucleus cellularity continued to decline throughout life whereas the annulus ceased its decline after the age of 50. Cell density was low in the vicinity of tears but elsewhere the disc was unaffected. Extensive inferior and superior end-plate separations reduced cellularity throughout the disc. Increased thickness of the cartilage end-plate was associated with higher cellularity in the nucleus.
Mesenchymal stem-cell based therapies have been
proposed as novel treatments for intervertebral disc degeneration,
a prevalent and disabling condition associated with back pain. The
development of these treatment strategies, however, has been hindered
by the incomplete understanding of the human nucleus pulposus phenotype
and by an inaccurate interpretation and translation of animal to
human research. This review summarises recent work characterising
the nucleus pulposus phenotype in different animal models and in
humans and integrates their findings with the anatomical and physiological
differences between these species. Understanding this phenotype
is paramount to guarantee that implanted cells restore the native
functions of the intervertebral disc. Cite this article:
We conducted a prospective follow-up MRI study
of originally asymptomatic healthy subjects to clarify the development
of Modic changes in the cervical spine over a ten-year period and
to identify related factors. Previously, 497 asymptomatic healthy
volunteers with no history of cervical trauma or surgery underwent
MRI. Of these, 223 underwent a second MRI at a mean follow-up of
11.6 years (10 to 12.7). These 223 subjects comprised 133 men and 100
women with a mean age at second MRI of 50.5 years (23 to 83). Modic
changes were classified as not present and types 1 to 3. Changes
in Modic types over time and relationships between Modic changes
and progression of degeneration of the disc or clinical symptoms
were evaluated. A total of 31 subjects (13.9%) showed Modic changes at
follow-up: type 1 in nine, type 2 in 18, type 3 in two, and types In the cervical spine over a ten-year period
We investigated the relationship between spinopelvic
parameters and
The December 2022 Spine Roundup. 360. looks at: Deep venous thrombosis prophylaxis protocol on a Level 1 trauma centre patient database; Non-specific spondylodiscitis: a new perspective for surgical treatment;
Aims. In this investigation, we administered oxidative stress to nucleus pulposus cells (NPCs), recognized DNA-damage-inducible transcript 4 (DDIT4) as a component in intervertebral disc degeneration (IVDD), and devised a hydrogel capable of conveying small interfering RNA (siRNA) to IVDD. Methods. An in vitro model for oxidative stress-induced injury in NPCs was developed to elucidate the mechanisms underlying the upregulation of DDIT4 expression, activation of the reactive oxygen species (ROS)-thioredoxin-interacting protein (TXNIP)-NLRP3 signalling pathway, and nucleus pulposus pyroptosis. Furthermore, the mechanism of action of small interfering DDIT4 (siDDIT4) on NPCs in vitro was validated. A triplex hydrogel named siDDIT4@G5-P-HA was created by adsorbing siDDIT4 onto fifth-generation polyamidoamine (PAMAM) dendrimer using van der Waals interactions, and then coating it with hyaluronic acid (HA). In addition, we established a rat puncture IVDD model to decipher the hydrogel’s mechanism in IVDD. Results. A correlation between DDIT4 expression levels and
The relationship of degeneration to symptoms has been questioned. MRI detects apparently similar
A key cause of low back pain is the degeneration of the intervertebral disc (IVD). Causality between infection of the IVD and its degenerative process gained great interest over the last decade. Granville Smith et al. (2021) identified 36 articles from 34 research studies investigating bacteria in human IVDs. Bacteria was identified in 27 studies, whereas 9 attributed bacterial presence to contamination. Cutibacterium acnes was the most abundant, followed by coagulase-negative staphylococcus. However, whether bacteria identified were present in vivo or represent perioperative contamination remains unclear. This study investigated whether bacteria are present in IVDs and what potential effects they may have on native disc cells. Immunohistochemical staining for Gram positive bacteria was performed on human IVD tissue to identify presence and characterise bacterial species. Nucleus pulposus (NP) cells in monolayer and 3D alginate were stimulated with LPS and Peptidoglycan (0.1-50 µg/ml) for 48hrs. Following stimulation qPCR for factors associated with
Bertolotti’s syndrome is characterised by anomalous enlargement of the transverse process(es) of the most caudal lumbar vertebra which may articulate or fuse with the sacrum or ilium and cause isolated L4/5 disc disease. We analysed the elective MR scans of the lumbosacral spine of 769 consecutive patients with low back pain taken between July 2003 and November 2004. Of these 568 showed
Osteoarthritis (OA) is a painful and disabling chronic condition that constitutes a major challenge to health care worldwide. There is currently no cure for OA and the analgesic pharmaceuticals available do not offer adequate and sustained pain relief, often being associated with significant undesirable side effects. Another disease associated with degenerating joints is Intervertebral disc degeneration (IVDD) which is a leading cause of chronic back pain and loss of function. It is characterized by the loss of extracellular matrix, specifically proteoglycan and collagen, tissue dehydration, fissure development and loss of disc height, inflammation, endplate sclerosis, cell death and hyperinnervation of nociceptive nerve fibers. The adult human IVD seems incapable of intrinsic repair and there are currently no proven treatments to prevent, stop or even retard
Objectives. Interleukin 18 (IL-18) is a regulatory cytokine that degrades the disc matrix. Bone morphogenetic protein-2 (BMP-2) stimulates synthesis of the disc extracellular matrix. However, the combined effects of BMP-2 and IL-18 on human intervertebral disc degeneration have not previously been reported. The aim of this study was to investigate the effects of the anabolic cytokine BMP-2 and the catabolic cytokine IL-18 on human nucleus pulposus (NP) and annulus fibrosus (AF) cells and, therefore, to identify potential therapeutic and clinical benefits of recombinant human (rh)BMP-2 in intervertebral disc degeneration. Methods. Levels of IL-18 were measured in the blood of patients with intervertebral disc degenerative disease and in control patients. Human NP and AF cells were cultured in a NP cell medium and treated with IL-18 or IL-18 plus BMP-2. mRNA levels of target genes were measured by real-time polymerase chain reaction, and protein levels of aggrecan, type II collagen, SOX6, and matrix metalloproteinase 13 (MMP13) were assessed by western blot analysis. Results. The serum level of patients (IL-18) increased significantly with the grade of IVD degeneration. There was a dramatic alteration in IL-18 level between the advanced degeneration (Grade III to V) group and the normal group (p = 0.008) Furthermore, IL-18 induced upregulation of the catabolic regulator MMP13 and downregulation of the anabolic regulators aggrecan, type II collagen, and SOX6 at 24 hours, contributing to degradation of disc matrix enzymes. However, BMP-2 antagonised the IL-18 induced upregulation of aggrecan, type II collagen, and SOX6, resulting in reversal of IL-18 mediated
Introduction and Objective. Intervertebral disc (IVD) degeneration accompanying with low back pain is a serious worldwide problem. Even though, surgical treatments are available for pain relief, there is an urgent need to establish enduring cell-based remedies. Notochordal (NC) cells as the ancestor of nucleus pulposus (NP) cells in human IVD are a promising therapeutic target. It has been reported that the loss of NC cells after childhood could promote the onset of
Aims. In order to evaluate the effectiveness of the Mobi-C implant
in cervical
Purpose of study and background. Mechanical overloading initiates intervertebral disc degeneration, presumably because cells break down the extracellular matrix (ECM). We used Fourier Transform Infrared Spectroscopy (FTIR) imaging to identify, visualize and quantify the ECM and aimed to identify spectroscopic markers for early
Aims. The aim of this study was to determine the differences in spinal imaging characteristics between subjects with or without lumbar developmental spinal stenosis (DSS) in a population-based cohort. Methods. This was a radiological analysis of 2,387 participants who underwent L1-S1 MRI. Means and ranges were calculated for age, sex, BMI, and MRI measurements. Anteroposterior (AP) vertebral canal diameters were used to differentiate those with DSS from controls. Other imaging parameters included vertebral body dimensions, spinal canal dimensions,
Biomechanical overloading initiates intervertebral disc degeneration. We hypothesized that this is due to mechanosensitivity of the cells, which break down the extracellular matrix. Previously, we found that overloading in a loaded disc culture system causes upregulation of remodeling- and inflammatory gene expressions. Fourier Transform Infrared Spectroscopy is a novel technique to identify, visualize and quantify ECM. In this research, we first identified novel spectroscopic markers for
Study design: Prospective clinical and radiologic study. Objective: The purpose of this study was to investigate the risk factors for adjacent segment degeneration after posterior lumbar interbody fusion (PLIF). Summary of Background data: Although several authors have reported the adjacent segment degeneration after lumbar or lumosacral fusion, there is no consensus regarding the risk factors for adjacent segment degeneration. Methods: Sixty-five patients were studied after PLIF after a minimum follow up time of 6 years. Plain and flexion/ extension radiographs and MRI scans were obtained and compared to preoperative and postoperative. Progression of segment degeneration was defined as a condition in which the distinction between nucleus and annulus is lost, and the disc space is collapsed according to the grading system (Grade 1–5) described by Pfirrmann et al evaluated with T2 weighted MRI scans. Patients were divided into three groups: Group 1 with no radiological progression of
Aims. Whether to perform hybrid surgery (HS) in contrast to anterior cervical discectomy and fusion (ACDF) when treating patients with multilevel cervical
Using deep learning and image processing technology, a standardized automatic quantitative analysis systerm of lumbar
Background. An improved understanding of intervertebral disc (IVD) structure and function is required for treatment development. Loading induces micro-fractures at the interface between the nucleus pulposus (NP) and the annulus fibrosus (AF), which is hypothesized to induce a cascade of cellular changes leading to degeneration. However, there is limited understanding of the structural relationship between the NP and AF at this interface and particularly response to load. Here, X-ray scattering is utilised to provide hierarchical morphometric information of collagen structure across the IVD, especially the interface region under load. Methodology. IVDs were imaged using the I22 SAXS/WAXS beamline at Diamond Light Source. Peaks associated with the D-banded structure of collagen fibrils were fitted to quantify their azimuthal distribution, as well the magnitude and direction of internal strains under static and applied strain (0–20%). Results. IVD tissue regions exhibited structural “AF-like” and “NP-like” fingerprints. Demonstrating high internal strains on collagen fibres particularly within the NP region of the disc. AF and NP regions showed distinct collagen orientation and internal strains with an apparent lack of bracing structure seen at the interface between the differential mechanical tissues. X-ray scattering under tensile strain provided structural information at high resolution, with clear differences observed between normal and degenerate discs under load. Conclusion. X ray scattering has been utilised to develop an improved understanding of collagen structure across the intervertebral disc which can be utilised to gain an increased understanding of load induced propagation of micro fissures and
Objectives. This study aims to investigate whether bacteria are present in intervertebral discs (IVDs) and their influence. Causality between chronic infection of the IVD and its degenerative process gained great interest recently. Granville Smith et al. (2021) identified 36 articles from 34 research studies investigating bacteria in IVDs, from these 27 studies found, Cutibacterium acnes being the most abundant. However, whether bacteria identified were present in vivo or if they represent contamination remains unclear. Methods. Human IVD tissue was fixed in paraffin and Immunohistochemical stained for Gram-positive bacteria. NP cells in monolayer have been stimulated with LPS (0.1–50 µg/ml) and Peptidoglycan (0.1–50 µg/ml) for 24, 48 and 72 hrs to investigate their influence. The concentration of proinflammatory and catabolic cytokines in the media is being measured using ELISA. RNA extracted and RT-qPCR utilised for factors associated with
Introduction: The effect of facet tropism on the development of lumbar disc diseases has been investigated but is still controversy; moreover, there has been no study to be done on far lateral lumbar disc herniation (LDH). In the current study, the authors attempted to determine the differences of the degree of facet tropism and the degree of
Lumbar diseases have become a major problem affecting human health worldwide. Conservative treatment of lumbar diseases is difficult to achieve ideal results, and surgical treatment of trauma, complications, it is imperative to develop a new treatment method. This study aims to explore the regulatory mechanism of cartilage endplate ossification caused by abnormal stress, and design intervention targets for this mechanism, so as to provide theoretical reference for the prevention and treatment of lumbar degeneration. In vivo, we constructed spinal instability model in mice. In vitro, we used a mechanical tensile machine to simulate the abnormal stress conditions of the endplate cartilage cells. Through the high-throughput sequencing, we found the enrichment of Hippo signaling pathway. As YAP is a key protein in the Hippo signaling pathway, we then created cartilaginous YAP elimination mice (Col2::YAPfl/fl). The lumbar spine model was constructed again in these mice for H&E, SOFG and immunofluorescence staining. In vitro lentivirus was used to knock out YAP, immunofluorescence staining, WB and qPCR were performed. Finally, we conducted therapeutic experiments by using YAP agonist and AAV5 carrying YAP plasmids. We collected 8w samples from C57/BL6 mice after modeling. We found ossification of the endplate in mice similar to human
Background. Magnetic resonance imaging (MRI) algorithm identifies end stage severely degenerated disc as ‘black’, and a moderately degenerate to non-degenerated disc as ‘white’. MRI is based on signal intensity changes that identifies loss of proteoglycans, water, and general radial bulging but lacks association with microscopic features such as fissure, endplate damage, persistent inflammatory catabolism that facilitates proteoglycan loss leading to ultimate collapse of annulus with neo-innervation and vascularization, as an indicator of pain. Thus, we propose a novel machine learning based imaging tool that combines quantifiable microscopic histopathological features with macroscopic signal intensities changes for hybrid assessment of
Anterior cervical discectomy and fusion (ACDF) is a well-established spinal operation for cervical
Invertebral
Aim of Study and Background. The vertebral endplate (VEP) is characterised as a bilayer of cartilage and bone, acting as a boundary between the disc and the vertebra. The disc being the largest avascular tissue in the body, relies primarily on the nutritional pathways from the vascular network in the adjacent VEP. Disruption of this nutrient supply has been identified as a major contributor to
Aims. The aim of this study was to determine the influence of developmental spinal stenosis (DSS) on the risk of re-operation at an adjacent level. Patients and Methods. This was a retrospective study of 235 consecutive patients who had undergone decompression-only surgery for lumbar spinal stenosis and had a minimum five-year follow-up. There were 106 female patients (45.1%) and 129 male patients (54.9%), with a mean age at surgery of 66.8 years (. sd. 11.3). We excluded those with adult deformity and spondylolisthesis. Presenting symptoms, levels operated on initially and at re-operation were studied. MRI measurements included the anteroposterior diameter of the bony spinal canal, the degree of
Clinical investigations show that the cervical spine presents wide inter-individual variability, where its motion patterns and load sharing strongly depend on the anatomy. The magnitude and scope of cervical diseases, including
Injury of the intervertebral disc (IVD) can occur for many reasons including structural weakness due to
Background. Intervertebral disc (IVD) degeneration is a major cause of low back pain (LBP). Degenerate discs are associated with accelerated cellular senescence. Cell senescence is associated with a secretory phenotype characterised by increased production of catabolic enzymes and cytokines. However to date, the mechanism of cell senescence within
Abstract. Objectives. While spinal fusion is known to be associated with adjacent
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
Intervertebral disc (IVD) degeneration is a major cause of low back pain (LBP). Degenerate discs are associated with accelerated cellular senescence. Cell senescence is associated with a secretory phenotype characterised by increased production of catabolic enzymes and cytokines. However, to date, the mechanism of cell senescence within
Introduction:
Despite a relentless search for adequate and effective treatment, low back pain is one of the most prevalent and costly illness in today’s society. While
Study Design and Objectives: The aim of this prospective study is to analyze clinical results of lumbar total disc arthroplasty according to the MRI evaluation of the
Introduction and Objective. Intervertebral disc (IVD) degeneration is one of the major contributors to low back pain, the leading cause of disability worldwide. This multifactorial pathological process involves the degradation of the extracellular matrix, inflammation, and cell loss due to apoptosis and senescence. While the deterioration of the extracellular matrix and cell loss lead to structural collapse of the IVD, increased levels of inflammation result in innervation and the development of pain. Amongst the known regulators of inflammation, toll-like receptors (TLRs) and more specifically TLR-2 have been shown to be specifically relevant in IVD degeneration. As strong post-transcriptional regulators, microRNAs (miRNAs) and their dysregulation has been connected to multiple pathologies, including degenerative diseases such as osteoarthritis and IVD degeneration. However, the role of miRNAs in TLR signalling in the IVD is still poorly understood and was hence investigated in this study. Materials and Methods. Human Nucleus pulposus (hNP) and Annulus fibrosus (hAF) cells (n=5) were treated with the TLR-2/6 specific agonist PAM2CSK4 (100 ng/mL for 6 hours) in order to activate the TLR2 signalling pathway. After the activation both miRNA and mRNA were isolated, followed by next-generation sequencing and qPCR analysis of proinflammatory cytokines respectively. Furthermore, cell supernatants were used to analyze the secretion of proinflammatory cytokines with enzyme-linked immunosorbent assay. TLR-2 knockdown (siRNA) cells were used as a control. Statistical analysis was conducted by performing Kolmogorov-Smirnov test and a two-tailed Student's t-test using GraphPad Prism version 9.0.2 for Windows (GraphPad Software, La Jolla California USA). Results. TLR-2 activation resulted in the induction of an inflammatory cell response, with a significant increase in gene expression of interleukin (IL)-6 (525 ± 180 fold change, p < 0.05) and IL-8 (7513 ± 1907 fold change, p < 0.05) and protein secretion of IL-6 (30.5 ± 8.1 pg/mL) and IL-8 (28.9 ± 5.4 pg/mL). TLR-2 activation was furthermore associated with changes in the miRNA profile of hNP and hAF cells. Specifically, we identified 10 differentially expressed miRNAs in response to TLR-2 activation, amongst which were miR-335–3p (1.45 log2 FC, p < 0.05), miR-125b-1–3p (0.55 log2 FC, p < 0.05), and miR-181a-3p (−1.05 log2 FC, p < 0.05). Conclusions. The identified miRNAs are known to be associated with osteoarthritis (miR-335-3p), inflammation and IVD degeneration (mir-125-1-3p and miR-181a-3p), but the link to TLR signalling has not been previously reported. Experiments to validate the identified miRNAs and elucidate their functional role are undergoing. The identification of these miRNAs provides an opportunity to further investigate miRNAs in the context of TLR activation and inflammation and to enhance our understanding of underlying molecular mechanisms behind
We have reviewed the cervical spine radiographs of 180 patients with athetoid cerebral palsy and compared them with those of 417 control subjects.
Purpose: Clinical practice guideline (CPG) concordant treatment (Ctx) has been shown to be more effective than CPG discordant care (Dtx) in a heterogeneous cohort of patients with acute lower back pain (ALBP). However, patients with underlying spine pathology (e.g. stenosis,
The basis of back pain and
Summary Statement. Conventional imaging techniques lack the ability to objectively assess early stages of intervertebral disc degeneration, characterised by glycosaminoglycan loss. This study shows that MRI T2∗ mapping correlates positively with GAG content and that it provides continuous measurements for
Background: Low back pain due to
Objectives. The aim of this study was to examine whether asymmetric loading
influences macrophage elastase (MMP12) expression in different parts
of a rat tail intervertebral disc and growth plate and if MMP12
expression is correlated with the severity of the deformity. Methods. A wedge deformity between the ninth and tenth tail vertebrae
was produced with an Ilizarov-type mini external fixator in 45 female
Wistar rats, matched for their age and weight. Three groups were
created according to the degree of deformity (10°, 30° and 50°).
A total of 30 discs and vertebrae were evaluated immunohistochemically
for immunolocalisation of MMP12 expression, and 15 discs were analysed
by western blot and zymography in order to detect pro- and active
MMP12. Results. No MMP12 expression was detected in the nucleus pulposus. Expression
of MMP12 in the annulus progressively increased from group I to
groups II and III, mainly at the concave side. Many growth plate
chondrocytes expressed MMP12 in the control group, less in group
I and rare in groups II and III. Changes in cell phenotype and reduction
of cell number were observed, together with disorganisation of matrix
microstructure similar to
We studied 52 patients, each with a lumbosacral transitional vertebra. Using MRI we found that the lumbar discs immediately above the transitional vertebra were significantly more degenerative and those between the transitional vertebrae and the sacrum were significantly less degenerative compared with discs at other levels. We also performed an anatomical study using 70 cadavers. We found that the iliolumbar ligament at the level immediately above the transitional vertebra was thinner and weaker than it was in cadavers without a lumbosacral transitional vertebra. Instability of the vertebral segment above the transitional vertebra because of a weak iliolumbar ligament could lead to subsequent
Introduction Intervertebral disc degeneration may cause chronic low back pain.
In our study, the aims were to describe the changes in the appearance of the lumbar spine on MRI in elite fast bowlers during a follow-up period of one year, and to determine whether these could be used to predict the presence of a stress fracture of the posterior elements. We recruited 28 elite fast bowlers with a mean age of 19 years (16 to 24) who were training and playing competitively at the start of the study. They underwent baseline MRI (season 1) and further scanning (season 2) after one year to assess the appearance of the lumbar intervertebral discs and posterior bony elements. The incidence of low back pain and the amount of playing and training time lost were also recorded. In total, 15 of the 28 participants (53.6%) showed signs of acute bone stress on either the season 1 or season 2 MR scans and there was a strong correlation between these findings and the later development of a stress fracture (p <
0.001). The prevalence of intervertebral disc degeneration was relatively low. There was no relationship between
As part of a 10 year follow-up study investigating the relationship between MRI-diagnosed disc disease and low back pain (LBP), a comparison of MRI image acquisition protocols was conducted. The aim was to establish whether the modern protocol produced improved diagnoses of lumbar disc disease. This is of significance when attempting to determine links between lumbar disc disease and LBP. The proposed hypothesis was that little difference in the pathology reported of MRI lumbar spines between the surface coil acquired images (Coil-MRI) and phased-array acquired images (Phased-MRI) would be found. Methods: Local ethics committee approval was granted for this study. 31 male subjects (aged 35–71 years) were recruited and underwent two subsequent scans. For both Coil-MRI and Phased-MRI scans sagittal dual echo, T1, axial T1 and T2 images were acquired. A Consultant Radiologist blindly reviewed the 62 scans continuously and reported on the pathology. Disease pathology assessment consisted of
Although success has been achieved with implantation of bone marrow mesenchymal stem cells (bMSCs) in degenerative discs, its full potential may not be achieved if the harsh environment of the degenerative disc remains. Axial distraction has been shown to increase hydration and nutrition. Combining both therapies may have a synergistic effect in reversing degenerative disc disease. In order to evaluate the effect of bMSC implantation, axial distraction and combination therapy in stimulating regeneration and retarding degeneration in degenerative discs, we first induced
Introduction Fusion has been the primary treatment for symptomatic
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
Introduction Cervical disc replacement has been advocated as a technique for retaining segmental motion following discectomy. The in vivo kinematics of the prosthesis are predicted to influence outcome, but remain poorly understood. The aim of this study was to determine whether factors that might possibly influence outcome can be reliably measured. Methods Anteroposterior and lateral xrays were performed pre-, inter- and post-operatively on 67 patients undergoing cervical prosthetic disc replacement. The xrays were reviewed by 3 observers. Measurements were performed manually by two observers and using digital image analysis software by a third observer. The inter-rater reliability of categorical measurements such as overall cervical alignment, degree of