Grade Of Intervertebral Disc Degeneration Research Articles

Relationship Between Vertebral Bone Marrow Edema and Early Progression of Intervertebral Disc Wedge or Narrowing After Lumbar Decompression Surgery.

A retrospective study. The aim of this study was to review clinical and imaging features in patients with lumbar spinal canal stenosis (LSS) with and without postoperative early progression of intervertebral disc degeneration (IDD) and to identify predictive factors. Progression of IDD after lumbar decompression surgery can induce low back pain and leg pain, and may require revision surgery. However, risk factors for postoperative radiological changes indicating IDD linked to development of symptoms have not been described. We included 564 patients with LSS without degenerative lumbar scoliosis who underwent lumbar decompression surgery without fusion. Clinical features and imaging findings were compared in cases with (group P) and without (group N) progression of IDD (intervertebral disc wedge or narrowing) at 1 year after surgery. Of the 564 patients, 49 (8.7%) were in group P. On preoperative MRI, all patients in group P had findings of vertebral bone marrow edema (diffuse high intensity on T2-weighted images and low-intensity on T1-weighted images), compared to only 5.4% in group N. The rate of revision surgery was significantly higher in group P in 5 years' follow-up (12.2% vs. 1.4%, P < 0.01). In group P, 44.9% of patients developed postoperative symptoms associated with postoperative radiological changes, and the frequency was higher in narrowing-type than in wedge-type cases. Vertebral bone marrow edema area and IDD grade were not predictors for postoperative early IDD progression. Careful consideration is required to determine whether lumbar decompression surgery should be performed if vertebral bone marrow edema is detected on MRI, since this is a predictor for a negative clinical outcome. If surgery is symptomati-cally urgent, careful clinical and radiological follow-up is required.Level of Evidence: 4.

Intervertebral disc degeneration relates to biomechanical changes of spinal ligaments

BACKGROUND CONTEXTThe ligamentum flavum (LF), the inter- and supraspinous ligament (ISL&SSL) and the intertransverse ligament (ITL) are relevant spinal structures for segmental stability. The biomechanical effect of degeneration and aging on their biomechanical properties remains largely unknown. PURPOSEThe aim of this study was to assess the material properties of the ITL, ISL&SSL and LF and to correlate parameters of biomechanical function with LF-thickness, intervertebral disc (IVD) degeneration and age. STUDY DESIGNBiomechanical cadaveric study. METHODSMRI- and CT-scans of 50 human lumbar segments (Th12-L5) were used to assess the ISL (acc. to Keorochana), the grade of IVD degeneration (acc. to Pfirrmann) and to quantify LF-thickness. The ITL, ISL&SSL and LF were resected in the neutral position of the spinal segment with a specifically developed method to conserve initial strain. Ramp to failure testing was performed (0.5 mm/s) to record initial tension, slack length, stiffness and ultimate strength. The relationship between the biomechanical characteristics and age and radiological parameters were analyzed. There are no study-specific conflicts of interest and no external funding was received for this study. RESULTSWith aging, a significant reduction in initial tension (r=-0.5, p<.01) and ultimate strength (r=-0.41, p<.01) of the LF was observed, while the effect on LF-stiffness and the characteristics of the other ligaments was non-significant. IVD-degeneration was correlated with a significant reduction in stiffness (r=-0.47, p=.001; r=-0.36, p=.01) and ultimate strength (r=-0.3, p=.04; r=-0.36, p=.01) of the LF and ISL&SSL respectively and a significant reduction in initial tension (r=-0.4, p<.01) of the LF. For the ITL, no significant correlation was observed. Comparing Pfirrman 2 to 5, this reduction was 40% to 80% for stiffness 60% to 70% for ultimate strength and 88% for initial tension of the LF. ISL&SSL-stiffness between Kerorochana grade A and D differed significantly (p=.03), while all other comparisons were non-significant (p>.05). LF-thickness did not correlate with the biomechanical properties of the LF (p>.05). CONCLUSIONSAging is primarily related to biomechanical changes to the LF. IVD-degeneration is related to a relevant reduction in stiffness and ultimate strength of the LF and ISL&SSL, with a similar trend for the ITL. The ISL-specific Keorochana grading system provides only minimal biomechanical information and LF-thickness does not provide biomechanical information. CLINICAL SIGNIFICANCEPatient age and the degenerative state of the IVD can be used to evaluate the biomechanical characteristics of the dorsal spinal ligaments, which can be helpful in selecting the optimal surgical procedure (e.g. in decompression surgery) for a specific situation.

Promoting Nrf2/Sirt3-Dependent Mitophagy Suppresses Apoptosis in Nucleus Pulposus Cells and Protects against Intervertebral Disc Degeneration.

One of the causes of intervertebral disc degeneration (IVDD) is nucleus pulposus cell (NPC) death, possibly apoptosis. In this study, we explored the role of the Nrf2/Sirt3 pathway and tert-butylhydroquinone (t-BHQ) in IVDD and elucidated the potential working mechanism. Reactive oxygen species (ROS) assay kits and malondialdehyde (MDA) assay kits were used to assess oxidative stress. Western blot and TUNEL staining were used to examine apoptosis. After siRNA against Nrf2 or lentivirus against Sirt3 was transfected into NPCs, the mechanism of the effect of the Nrf2/Sirt3 pathway on NPCs was assessed. The interaction between t-BHQ and its potential interacting protein NRF2 was further investigated through protein docking analysis. ChIP examined the binding affinity between Nrf2 and Sirt3 promoter. In vivo experiments, X-ray, hematoxylin-eosin (HE) staining, Safranin O staining, and immunohistochemistry were used to evaluate IVDD grades. The results demonstrated that activation of the Nrf2/Sirt3 pathway inhibited tert-butyl hydroperoxide- (TBHP-) induced apoptosis and mitochondrial dysfunction in vitro. In addition to apoptosis, upregulation of the Nrf2/Sirt3 pathway induced by t-BHQ restored TBHP-induced autophagic flux disturbances. However, its protective effect was reversed by chloroquine and Si-ATG5. Furthermore, t-BHQ ameliorated IVDD development in a rat model. In conclusion, our findings indicate that the Nrf2/Sirt3 pathway and its agonist represent a potential candidate for treating IVDD.

Effect of posterior lumbar fusion on the degeneration of adjacent segments using cortical bone trajectory screw fixation

Objective: To compare the degenerative changes of adjacent segments after posterior lumbar fusion surgery using cortical bone trajectory (CBT) screws and pedicle screws (PS) in lumbar spinal degenerative disease. Methods: According to certain inclusion and exclusion criteria, prospective non-randomized study on cases of the single-segment lumbar spinal fusion surgery using cortical bone trajectory screws fixation and pedicle screws fixation in Beijing Chao-Yang Hospital from January 2015 to February 2016 was performed. The intervertebral space height (ISH), foramen height (FH), foramen width (FW), cephalad adjacent segment (CAS) and intervertebral disc degeneration grades at MRI scans were measured before surgery and during follow-up. Clinical symptoms were evaluated by Oswestry Dysfunction Index (ODI) score and pain visual analogue scale (VAS) before and during the follow-up. Radiologic adjacent segment degeneration (R-ASD) and symptomatic adjacent segment disease (S-ASD) patients were diagnosed during the follow-up. Paired sample t test was performed when data were compared before surgery and during follow-up. Results: A total of 69 patients were included in the study, 33 in the CBT group (male/female, 15/18), with an average age of (65±11) years, and 36 in the PS group (male/female, 17/19), with an average age of (64±10) years. The follow-up time was no less than 36 months. At the last follow-up, the ISH of the adjacent segments in the CBT group were not statistically different from that before surgery; the ISH of the adjacent segments in the PS group were significantly reduced compared with that before surgery (t=6.78, P<0.05). The loss of ISH, FH, and FW in the adjacent segments in the CBT group were smaller than those in the PS group, and the differences were statistically significant. During follow-up, 4 cases (12.1%) of R-ASD and no S-ASD were found in the CBT group, while 12 cases (33.3%) of R-ASD appeared in the PS group, which was significantly higher than that in the CBT group (χ(2)=4.35, P=0.04). According to MRI, the adjacent discs of PS group had significantly more severe degeneration at the last follow-up than the CBT group (Z=-2.14, P=0.03). Conclusions: Compared with the PS fixation fusion, the CBT screw fixation can effectively reduce the occurrence of adjacent segment degeneration.

MRI evaluation of degenerative lumbar spine disease in young adults : a quantitative evaluation of risk factors.

Background Data: Evidently, there is an alarmingly increasing incidence of degenerative lumbar spine disease in adolescents and young adults. Hence, it is imperative to ascertain and identify the risk factors so that early lifestyle modifications can be done, and early lumbar spine degeneration can be prevented. Purpose: To study the array of MRI findings of degenerative lumbar spine disease in young adults and quantitatively evaluate the risk factors. Study Design: A prospective imaging case study.Patients and Methods: A total of 200 young adult patients between 21 and 30 years old presenting with low back pain were included in this study. MRI findings of 200 young adults were graded and quantitatively evaluated in relation to three risk factors, that is, physical activity, smoking, and BMI. Grading of intervertebral disc degeneration, lumbar facet joint degeneration, and foraminal stenosis was done.Results: Modic type II endplate changes and reduced T2 bright disc signal were the most common manifestations of spondylosis and intervertebral disc (IVD) degeneration, respectively. A correlation between obesity and increasing grades of disc degeneration was found to be most significant at a 95% confidence interval. A significant correlation was found between each of the risk factors (obesity, sedentary lifestyle, and smoking) and increasing grades of IVD degeneration and facet arthropathy with a p value of <0.05. Foraminal stenosis manifested a positive association with BMI at a 95 % confidence interval. Conclusion: This study could recognize a statistically significant correlation between lumbar DDD and physical activity, smoking, and BMI. Identification of causative lifestyle factors could help prevent the occurrence and progression of lumbar DDD. .

Differential regulation of TRP channel gene and protein expression by intervertebral disc degeneration and back pain

Intervertebral disc (IVD) degeneration and consequent low back pain (LBP) are common and costly pathological processes that require improved treatment strategies. Transient Receptor Potential (TRP) channels constitute a family of multimodal ion channels that have recently emerged as contributors to disc pathologies and were thus proposed as potential therapeutic targets, although limited data on their presence and function in the IVD exist. The purpose of this study was to determine the mRNA and protein expression of TRP channels in non-degenerated and degenerated human IVD tissue (with different pain intensity and chronicity) using gene array, conventional qPCR and immunohistochemistry. We could demonstrate that 26 out of 28 currently known TRP channels are expressed in the IVD on the mRNA level, thereby revealing novel therapeutic candidates from the TRPC, TRPM and TRPML subfamilies. TRPC6, TRPM2 and TRPML1 displayed enhanced gene and protein expression in degenerated IVDs as compared to non-degenerated IVDs. Additionally, the gene expression of TRPC6 and TRPML1 was influenced by the IVD degeneration grade. Pain intensity and/or chronicity influenced the gene and/or protein expression of TRPC6, TRPM2 and TRML1. Interestingly, decreased gene expression of TRPM2 was observed in patients treated with steroids. This study supports the importance of TRP channels in IVD homeostasis and pathology and their possible application as pharmacological targets for the treatment of IVD degeneration and LBP. However, the exact function and activation of the highlighted TRP channels will have to be determined in future studies.

Melatonin modulates IL-1β-induced extracellular matrix remodeling in human nucleus pulposus cells and attenuates rat intervertebral disc degeneration and inflammation.

The inflammatory-associated factors interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) are widely reported to be associated with intervertebral disc (IVD) degeneration (IVDD). N-acetyl-5-methoxytryptamine (melatonin) is a natural hormone secreted by the pineal gland which has been shown to participate in several physiological and pathological progresses, such as aging, anti-inflammation, anti-apoptosis and autophagy regulation. However, the effects of melatonin on IVD remain unclear. In the present study, we treated human nucleus pulposus cells (NPCs) with melatonin and discovered that melatonin could modulate extracellular matrix (ECM) remodeling induced by IL-1β by enhancing collagen II and aggrecan expression levels and by downregulating matrix metalloproteinase-3 (MMP-3) levels. These findings were verified by western blot and immunofluorescence assays. Intraperitoneal injection of melatonin mitigated IVDD in the rat tail puncture model. X-ray and magnetic resonance imaging (MRI), as well as hematoxylin-eosin (H&E), Safranine O-Green, Alcian blue and Celium red staining methods were adopted to evaluate IVDD grades, the structural integrity of nucleus pulposus (NP) and annulus fibrosus (AF) and the damage and calcification of the cartilage endplate. Melatonin reduced inflammatory cell aggregation and the release of the inflammatory factors IL-1β, IL-6, TNF-α as determined by immunohistochemistry. In conclusion, the present study demonstrated that melatonin could modulate ECM remodeling by IL-1β in vitro and attenuate the IVDD and induction of inflammation in a rat tail puncture model in vivo. The data demonstrated that melatonin may contribute to the restoration processs of IVD following damage and may be used as a potential novel therapy for IVDD.

Thoracic spinal kinematics is affected by the grade of intervertebral disc degeneration, but not by the presence of the ribs: An in vitro study

BACKGROUND CONTEXTThoracic spinal three-dimensional kinematics is widely unknown. For the evaluation of surgical treatments and the complete validation of numerical models, however, kinematic data of the thoracic spine are essential. PURPOSETo identify possible effects of rib presence and grade of intervertebral disc degeneration on thoracic spinal kinematics including three-plane helical axes and instantaneous centers of rotation. DESIGN/SETTINGRadiological grading of intervertebral disc degeneration and in vitro tests using n=8 human thoracic functional spinal units of the segmental levels T1–T2, T3–T4, T5–T6, T7–T8, T9–T10, and T11–T12, respectively, were performed with as well as without ribs to analyze the specific kinematic properties. METHODSSpecimens were loaded with pure moments of 5 Nm and constant loading rates of 1°/s in flexion/extension, lateral bending, and axial rotation. Optical motion tracking was performed to visualize helical axes and instantaneous centers of rotation on three-plane X-rays and to evaluate primary ranges of motion (ROMs) and coupled motions. RESULTSMotion segments with no or mild disc degeneration showed reproducible kinematics in all motion planes, whereas medium or severely degenerated specimens offered high variations and shifts of the rotational axes to the distal direction as well as lower ROM. Coupled motions were generally not detected. CONCLUSIONSWith progressing disc degeneration, the rotational axes show higher variation and tend to shift in distal direction, especially in flexion/extension with a shift to the anterior direction, whereas rib resection does not affect thoracic spinal kinematics but its stability. Rib resections as part of spinal deformity treatment destabilize the thoracic spine, but do not alter its kinematics. Young and healthy discs, however, could be affected by surgical treatments of the thoracic spine regarding thoracic spinal kinematics.

Kinsenoside ameliorates intervertebral disc degeneration through the activation of AKT-ERK1/2-Nrf2 signaling pathway.

Intervertebral disc degeneration (IDD) is recognized as the major contributor to low back pain, which results in disability worldwide and heavy burdens on society and economy. Here we present evidence that the lower level of Nrf2 is closely associated with higher grade of IDD. The apoptosis and senescence of nucleus pulposus cells (NPCs) were exacerbated by Nrf2 knockdown, but suppressed by Nrf2 overexpression under oxidative stress. Based on findings that Kinsenoside could exert multiple pharmacological effects, we found that Kinsenoside rescued the NPC viability under oxidative stress and protected against apoptosis, senescence and mitochondrial dysfunction in a Nrf2-dependent way. Further experiments revealed that Kinsenoside activated a signaling pathway of AKT-ERK1/2-Nrf2 in NPCs. Moreover, in vivo study showed that Kinsenoside ameliorated IDD in a puncture-induced model. Together, the present work suggests that Nrf2 is involved in the pathogenesis of IDD and shows the protective effects as well as the underlying mechanism of Kinsenoside on Nrf2 activation in NPCs.

LncRNA HOTAIR upregulates autophagy to promote apoptosis and senescence of nucleus pulposus cells.

Intervertebral disc degeneration (IDD) is a complex and chronic disease that involves disc cell senescence, death, and extracellular matrix (ECM) degradation. HOTAIR, a long non-coding RNA (lncRNA) is reportedly associated with autophagy, whereas autophagy is shown to promote IDD. However, how it affects nucleus pulposus (NP) cells, the primary component of intervertebral discs is still unclear. We hypothesized that HOTAIR promotes NP cell apoptosis and senescence through upregulating autophagy. Thus, silencing HOTAIR should inhibit autophagy and exert a therapeutic effect on IDD. Our in vitro experiments in human NP cells revealed that HOTAIR expression positively correlated with IDD grade, and overexpression enhanced autophagy. Autophagy inhibition via 3-methyladenine reversed HOTAIR stimulatory effects on apoptosis, senescence, and ECM catabolism, while the AMP-activated protein kinase (AMPK) inhibitor Compound C suppressed HOTAIR-induced autophagy through regulating AMPK/mTOR/ULK1 pathways. Our in vivo experiment then illustrated that silencing HOTAIR ameliorates IDD in rats. Collectively, we demonstrated that HOTAIR stimulates autophagy to promote NP cell apoptosis, senescence, and ECM catabolism. Therefore, silencing HOTAIR has the potential to become a treatment option for IDD.

Angiopoietin-like protein 8 expression and association with extracellular matrix metabolism and inflammation during intervertebral disc degeneration.

Intervertebral disc degeneration (IDD) is considered the primary culprit for low back pain. Although the underlying mechanisms remain unknown, hyperactive catabolism of the extracellular matrix (ECM) and inflammation are suggested to play critical roles in IDD progression. This study was designed to elucidate the role of angiopoietin‐like protein 8 (ANGPTL8) in the progression of IDD, especially the relationship of ANGPTL8 with ECM metabolism and inflammation. A positive association between ANGPTL8 expression and degenerative grades of IDD was detected in the analysis of human nucleus pulposus tissue samples. Silencing of ANGPTL8 attenuated the degradation of the anabolic protein type collagen II, and reduced the expression of the catabolic proteins MMP3 and MMP9, and the inflammatory cytokine IL‐6 through inhibition of NF‐κB signalling activation. In addition, the effect of ANGPTL8 was evaluated in a rat model of puncture‐induced IDD. Based on the imaging results and histological examination in animal study, knockdown of ANGPTL8 was demonstrated to ameliorate the IDD progression. These results demonstrate the detrimental role of ANGPTL8 expression in the pathogenesis of IDD and may provide a new therapeutic target for IDD treatment.

TFEB protects nucleus pulposus cells against apoptosis and senescence via restoring autophagic flux

Excessive apoptosis and senescence of nucleus pulposus (NP) cells are major pathological changes in intervertebral disc degeneration (IVDD) development; previous studies demonstrated pharmacologically or genetically stimulation of autophagy may inhibit apoptosis and senescence in NP cells. Transcription factor EB (TFEB) is a master regulator of autophagic flux via initiating autophagy-related genes and lysosomal biogenesis. This study was performed to confirm whether TFEB was involved in IVDD development and its mechanism. TFEB activity was detected in NP tissues in puncture-induced rat IVDD model by immunofluorescence as well as in tert-Butyl hydroperoxide (TBHP), the reactive oxygen species (ROS) donor to induce oxidative stress, treated NP cells by western blot. After TFEB overexpression in NP cells with lentivirus transfection, autophagic flux, apoptosis and senescence percentage were assessed. In invivo study, the lentivirus-normal control (LV-NC) or lentivirus-TFEB (LV-TFEB) were injected into the center space of the NP tissue, after 4 or 8 weeks, Magnetic resonance imaging (MRI), X ray, Hematoxylin-Eosin (HE) and Safranin O staining were used to evaluate IVDD grades. The nuclear localization of TFEB declined in degenerated rat NP tissue as well as in TBHP treated NP cells. Applying lentivirus to transfect NP cells, TFEB overexpression restored the TBHP-induced autophagic flux blockage and protected NP cells against apoptosis and senescence; these protections of TFEB are diminished by chloroquine-medicated autophagy inhibition. Furthermore, TFEB overexpression ameliorates the puncture-induced IVDD development in rats. Experimental IVDD inhibited the TFEB activity. TFEB overexpression suppressed TBHP-induced apoptosis and senescence via autophagic flux stimulation in NP cell and alleviates puncture-induced IVDD development invivo.

CircularRNA_104670 plays a critical role in intervertebral disc degeneration by functioning as a ceRNA

This study was carried out to explore the roles of circular RNAs (circRNAs) in nucleus pulposus (NP) tissues in intervertebral disc degeneration (IDD). Differentially expressed circRNAs in IDD and normal NP tissues were identified based on the results of microarray analysis. Bioinformatics techniques were employed to predict the direct interactions of selected circRNAs, microRNAs (miR), and mRNAs. CircRNA_104670 was selected as the target circRNA due to its large multiplier expression in IDD tissues. After luciferase reporter and EGFP/RFP reporter assays, we confirmed that circRNA_104670 directly bound to miR-17-3p, while MMP-2 was the direct target of miR-17-3p. The receiver-operating characteristic (ROC) curve showed that circRNA_104670 and miR-17-3p had good diagnostic significance for IDD (AUC circRNA_104670 = 0.96; AUC miRNA-17-3p = 0.91). A significant correlation was detected between the Pfirrmann grade and expression of circRNA_104670 (r = 0.63; p = 0.00) and miR-17-3p (r = −0.62; p = 0.00). Flow-cytometric analysis and the MTT assay showed that interfering with circRNA_104670 using small interfering RNA (siRNA) inhibited NP cell apoptosis (p < 0.01), and this inhibition was reduced by interfering with miR-17-3p. Interfering with circRNA_104670 suppressed MMP-2 expression and increased extracellular matrix (ECM) formation, which were also reduced by interfering with miR-17-3p. Finally, an MRI evaluation showed that circRNA_104670 inhibition mice had a lower IDD grade compared with control mice (p < 0.01), whereas circRNA_104670 and miRNA-17-3p inhibition mice had a higher IDD grade compared with circRNA_104670 inhibition mice (p < 0.05). CircRNA_104670 is highly expressed in the NP tissues of IDD and acts as a ceRNA during NP degradation.

Clinical Relationship of Degenerative Changes between the Cervical and Lumbar Spine.

Study DesignRetrospective, observational, case series.PurposeTo elucidate the prevalence of degenerative changes in the cervical and lumbar spine and estimate the degenerative changes in the cervical spine based on the degeneration of lumbar disc through a retrospective review of magnetic resonance (MR) images.Overview of LiteratureOver 50% of middle-aged adults show evidence of spinal degeneration. However, the relationship between degenerative changes in the cervical and lumbar spine has yet to be elucidated.MethodsA retrospective review of positional MR images of 152 patients with symptoms related to cervical and lumbar spondylosis with or without a neurogenic component was conducted. The degree of intervertebral disc degeneration (IDD) was assessed on a grade of 1–5 for each segment of the cervical and lumbar spine using MR T2-weighted sagittal images. The grades across all segments were summed to produce the degenerative disc score (DDS) for the cervical and lumbar spine. The patients were divided into two groups based on the IDD grade for each lumbar segment: normal (grades 1 and 2) and degenerative (grades 3–5).ResultsDDSs for the cervical and lumbar spine were positively correlated. Significant differences in cervical DDSs between the groups were observed in all lumbar segments. Although there were no significant differences in cervical DDSs among the degenerative lumbar segment, cervical DDSs at the L1–2 and L2–3 segments tended to be higher than those at the L3–4, L4–5, and L5–S degenerative segments.ConclusionsOur study shows that participants with degenerative changes in the upper lumbar segments are more likely to have a certain amount of cervical spondylosis. This information could be used to lower the incidence of a missed diagnosis of cervical spine disorders in patients presenting with lumbar spine symptomology.

Intervertebral Disc Degeneration in Warmblood Horses: Morphology, Grading, and Distribution of Lesions.

Equine intervertebral disc degeneration is thought to be rare and of limited clinical relevance, although research is lacking. To objectively assess pathological changes of the equine intervertebral disc and their clinical relevance, description of the normal morphology and a practical, biologically credible grading scheme are needed. The objectives of this study are to describe the gross and histological appearance of the equine intervertebral discs and to propose a grading scheme for macroscopic degeneration. Spinal units from 33 warmblood horses were grossly analyzed and scored. Of the 286 intervertebral discs analyzed, 107 (37%) were assigned grade 1 and grade 2 (considered normal) and were analyzed histologically. A nucleus pulposus and an annulus fibrosus could be identified macroscopically and histologically. Histologically, the nucleus pulposus was composed of a cartilaginous matrix and the annulus fibrosus of parallel collagenous bands. A transition zone was also histologically visible. Intra- and inter-observer reliability scores were high for all observers. Higher grades were associated with greater age. Gross changes associated with equine intervertebral disc degeneration (grades 3-5)-that is, yellow discoloration, cleft formation (tearing), and changes in consistency of the nucleus pulposus-were largely similar to those in humans and dogs and were most prevalent in the caudal cervical spine. Equine intervertebral disc degeneration was not associated with osteophyte formation. Changes of the vertebral bone were most common in the thoracolumbar spine but were not correlated with higher grades of intervertebral disc degeneration. Thus, changes of the vertebral bone should be excluded from grading for equine intervertebral disc degeneration.

Dysregulated MiR-3150a-3p Promotes Lumbar Intervertebral Disc Degeneration by Targeting Aggrecan

Background/Aims: Low back pain has become one of the most common musculoskeletal diseases in the world. Studies have shown that intervertebral disc degeneration (IDD) is an important factor leading to low back pain, but the mechanisms underlying IDD remain largely unknown. Research over the past decade has suggested critical roles for microRNAs (miRNAs) in natural growth and disease progression. However, it remains poorly understood whether circular RNAs participate in IDD. Methods: Clinical IDD samples were collected from 20 patients who underwent discectomy. Weighted gene co-expression network analysis was used to identify the co-expression miRNA network modules (highly co-expressed clusters of miRNAs) that were associated with IDD grade. Results: miR-3150a-3p was the most significantly up-regulated miRNA in module “Blue.” Notably, aggrecan (ACAN) was identified as a direct target gene of miR-3150a-3p and ACAN expression was regulated by miR-3150a-3p. Overexpression of miR-3150a-3p decreased ACAN expression in nucleus pulposus cells, whereas inhibition of miR-3150a-3p increased ACAN expression. In addition, ACAN expression was negatively correlated with IDD grade. Conclusion: Our study suggests that the reduction of ACAN expression induced by the upregulation of miR-3150a-3p might participate in the development of IDD.

Nicotinamide Phosphoribosyltransferase Inhibitor APO866 Prevents IL-1β-Induced Human Nucleus Pulposus Cell Degeneration via Autophagy

Background/Aims: Intervertebral discs consist of an extracellular matrix (ECM) with a central gelatinous nucleus pulposus (NP) enclosed in an outer layer known as the annulus fibrosus. ECM metabolic disorders result in loss of boundary between the annulus fibrosus and NP, which can lead to intervertebral disc degeneration (IDD). Proinflammatory cytokines, such as interleukin (IL)-1β, mediate the progression of IDD. Nicotinamide phosphoribosyltransferase (Nampt) catalyzes the first step in the biosynthesis of nicotinamide adenine dinucleotide (NAD) and is known to be induced by IL-1β. APO866 is an inhibitor of NAD biosynthesis and is involved in autophagy. LC3 (microtubule-associated protein 1 light chain 3) is a key regulator of autophagy and is used as an indicator of increased autophagy. Herein, we investigate the role of APO866 in regulating autophagy in NP cells and IL-1β mediated NP cell degeneration and apoptosis. Methods: NP cells were extracted from IDD tissues and cultured in DMEM/F12 medium. Nampt was induced by different concentrations of IL-1β (0, 0.5, 1, 5, 10 ng/mL) for 24 h or NP cells were treated with 10 ng/mL IL-1β for 0, 6, 12, 48 h. QRT-PCR and western blots were used to detect Nampt and ECM-related protein expression in NP tissue of patients with IDD and in NP cells. Confocal analysis was used to detect membrane-bound LC3, Aggrecan, and Collagen II. Results: Nampt is expressed in NP tissue at higher levels in severe grades of IDD (Grade IV and V) compared with low grades (Grade II and III). In NP cells, 10 ng/mL IL-1β induced Nampt expression for 48 h, increased expression of the degradative-associated proteins, ADAMTS4/5 and MMP-3/13, and decreased expression of ECM-related proteins, Aggrecan and Collagen II. However, the Nampt inhibitor APO866 blocked IL-1β induction, and the knockdown of Nampt expression increased the expression of ECM proteins that were inhibited by IL-1β. Moreover, evidence provided by the autophagic markers LC3 and Beclin-1 indicated that APO866 induced NP cell autophagy. Furthermore, although APO866 inhibited the downregulated expression of ECM-related proteins by IL-1β, this function was blocked by autophagy inhibitor, 3-methyladenine. Conclusion: APO866 protects NP cells and induces autophagy by inhibiting IL-1β-induced NP cell degeneration and apoptosis, which may have therapeutic potential in IDD.

Association between intervertebral disc degeneration and the Oswestry Disability Index.

Low back pain and lumbar intervertebral disc degeneration (IDD) are common findings. Valid data on correlation between clinical pain scores and grades of IDD are not available. To investigate the correlation of intervertebral disc degeneration (IDD) at lumbar levels L4/5 and L5/S1 and the Oswestry Disability Index (ODI). The lumbar discs L4/5 and L5/S1 of 591 patients were evaluated according to the 5-point (Grade I to Grade V) grading system as published by Pfirrmann et al. Functional status was assessed using the Oswestry Disability Index. Spearman's coefficient of rank correlation was used for statistical analysis (p < 0.05). The majority of patients revealed lumbar discs with Pfirrmann grade II to grade IV changes (93.3% at level L4/5; 89.8% at level L5/S1), while a relatively low percentage of lumbar discs presented with grade I (level L4/5: 1.5%; level L5/S1: 2.0%) or grade V (level L4/5: 5.1%; level L5/S1: 8.1%) changes, respectively. Patients' ODI scores ranged between 0 and 91.11% (arithmetic mean of 32.77% ± 17.02%). The largest group of patients (48.39%) had moderate functional disability (ODI score between 21 and 40%). There was a weak, but statistically significant positive correlation between IDD and ODI for both evaluated lumbar levels. Increased lumbar IDD in MRI goes along with an increased ODI. Thus, MRI is a strong indicator of a patient's clinical appearance. However, low back pain cannot be explained by imaging alone. Clinical correlation is imperative for an adequate diagnostic advance in patients with low back pain.

Self-complementary adeno-associated virus serotype 6 mediated knockdown of ADAMTS4 induces long-term and effective enhancement of aggrecan in degenerative human nucleus pulposus cells: A new therapeutic approach for intervertebral disc disorders.

Inhibition of intervertebral disc (IVD) degeneration, which is often accompanied by painful inflammatory and immunopathological processes, is challenging. Current IVD gene therapeutic approaches are based on adenoviral gene delivery systems, which are limited by immune reactions to their viral proteins. Their applications in IVDs near to sensitive neural structure could provoke toxicity and immunological side-effects with neurological deficits. Self-complementary adeno-associated virus (scAAV) vectors, which do not express any viral gene and are not linked with any known disease in humans, are attractive therapeutic gene delivery vectors in degenerative IVDs. However, scAAV-based silencing of catabolic or inflammatory factor has not yet been investigated in human IVD cells. Therefore, we used scAAV6, the most suitable serotype for transduction of human nucleus pulposus (NP) cells, to knockdown the major catabolic gene (ADAMTS4) of IVD degeneration. IVD degeneration grades were determined by preoperative magnetic resonance imaging. Lumbar NP tissues of degeneration grade III were removed from 12 patients by nucleotomy. NP cells were isolated and cultured with low-glucose. Titre of recombinant scAAV6 vectors targeting ADAMTS4, transduction efficiencies, transduction units, cell viabilities and expression levels of target genes were analysed using quantitative PCR, fluorescence microscopy, fluorescence-activated cell sorting, 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide assays, quantitative reverse transcription PCR, western blot and enzyme-linked immunosorbent assays during 48 days of post-transduction. Transduction efficiencies between 98.2% and 37.4% and transduction units between 611 and 245 TU/cell were verified during 48 days of post-transduction (p<0.001). scAAV6-mediated knockdown of ADAMTS4 with maximum 87.7% and minimum 40.1% was confirmed on day 8 and 48 with enhanced the level of aggrecan 48.5% and 30.2% respectively (p<0.001). scAAV6-mediated knockdown of ADAMTS4 showed no impact on cell viability and expression levels of other inflammatory catabolic proteins. Thus, our results are promising and may help to design long-term and less immunogenic gene therapeutic approaches in IVD disorders, which usually need prolonged therapeutic period between weeks and months.

Relation between grades of intervertebral disc degeneration and occupational activities of patients with lumbar disc herniation

Background/Aim. Intervertebral disc degeneration (IDD) occurs as consequence of combined effects of genetic, age-related, environmental and occupational factors. Lumbar disc herniation (LDH) develops mostly due to IDD. The aim of this study was to investigate whether the frequency of LDH is higher at the level of the most pronounced IDD, and whether a category of physical workload influences higher IDD on level L3-L4, L4-L5 and L5-S1 separately. Methods. The research included 60 patients with permanent employment, hospitalized due to LDH. A grade of IDD was assessed by lumbosacral preoperative magnetic resonance imaging (MRI), according to Pfirmann's MRI classification system. Occupational factors were determined by a specific questionnaire. Results. Out of the 60 patients participating in the study, 33.3% had jobs with easy workload, 23% had moderate workload, while 43% had heavy workload. Herniated discs were found at level L3-L4 in 8.3%, at level L4-L5 in 46.7% and at level L5-S1 in 45% patients. The symptomatic discs at level L5-S1 showed statistically significant frequency of degenerative changes of grades IV and V. Binary logistic regression results showed that the strongest predictor of IDD grade for examined levels was physical workload. Positive association of physical workload and IDD grade was observed in all cases. Higher grades of IDD are more likely for patients with both higher TE and heavier physical workload (OR 2.011) at level L3-L4. At levels L4-L5 and L5-S1 higher degree of IDD was more likely for females with heavier physical workload (OR 1.978 and 2.433 respectively). Conclusion. Symptomatic discs show higher frequency of higher grades of IDD but herniation does not occur solely at the disc of the most prominent degenerative changes. The results suggest importance of inter-influence of physical workload and the years of employment and the inter-influence of physical workload and gender, on degeneration of lumbar discs.