Degenerative Changes In Intervertebral Discs Research Articles

Andrographolide influences IDD cell autophagy and oxidative stress under mechanical pressure via miR-9/FoxO3/PINK1/Parkin molecular axis.

Intervertebral disc degeneration (IDD) is characterized by the decreased function and number of nucleus pulposus cells (NPCs) caused by excessive intervertebral disc (IVD) pressure. This research aims to provide novel insights into IDD prevention and treatment by clarifying the effect of andrographolide (ANDR) on IDD cell autophagy and oxidative stress under mechanical stress. Human primary NPCs were extracted from the nucleus pulposus tissue of non-IDD trauma patients. An IDD cell model was established by posing mechanical traction on NPCs. Through the construction of an IDD rat model, the influence of ANDR on IDD pathological changes was explored in vivo. The proliferation and autophagy of NPCs were decreased while the apoptosis rate and oxidative stress reaction were increased by mechanical traction. ANDR intervention obviously alleviated this situation. MiR-9 showed upregulated expression in IDD cell model, while FoxO3 and PINK1/Parkin were downregulated. Decreased proliferation and autophagy as well as enhanced apoptosis and oxidative stress response of NPCs were observed following miR-9 mimics and H89 intervention, while the opposite trend was observed after FoxO3 overexpression. FoxO3 is a direct target downstream miR-9. The in vivo experiments revealed that after ANDR intervention, the number of apoptotic cells in rat IVD tissue decreased and the autophagy increased. In conclusion, ANDR improves NPC proliferation, and autophagy, inhibits apoptosis and oxidative stress, and alleviates the pathological changes of IDD via the miR-9/FoxO3/PINK1/Parkin axis, which may be a new and effective treatment for IDD in the future.

Efficacy of wet cupping in the pain management of cervical spondylosis – A randomized, controlled, open -label, parallel-group clinical trial

Background and objectiveCervical spondylosis is a chronic degenerative process of the cervical spine characterized by pain in neck, degenerative changes in intervertebral disc and osteophyte formation. Cervical spondylosis is translated as Wajaʻ ur Raqaba, a type of joint pain. The present study was aimed to evaluate the effect of wet cupping in the pain management of cervical spondylosis. MethodsThis Open, randomized, controlled, clinical study was conducted on 44 patients. Subjects in the test group (n = 22) received a series of three-staged wet cupping treatment, performed on 0, 7th and 14th day. Subjects in the control group (n = 22) received 12 sittings of Transcutaneous Electrical Nerve Stimulation (TENS): 6 sittings per week for two weeks. The objective findings of treatment were assessed with the help of VAS, Neck Disability Index (NDI) and Cervical range of motion. ResultsIntra group comparison in test group from baseline to 21st day were found highly significant (p < 0.001) in terms of VAS, NDI, Flexion, Extension and Left rotation score. While in Right rotation, Left rotation and Left lateral flexion score were found moderately significant (p < 0.01). Statistically significant difference was observed between two groups at 21st day in VAS scale, NDI, and Cervical range of motion score (p < 0.001). Interpretation and conclusionḤijāma Bish Sharṭ was found better in the management of pain due to cervical spondylosis than TENS. It can be concluded that Ḥijāma Bish Sharṭ may a better option for the pain management of cervical spondylosis. Clinical trial registrationThe trial was registered on clinical trial registry website (www.ctri.nic.in) bearing a CTRI Number, CTRI/2020/03/024,249.

Sinapine thiocyanate alleviates intervertebral disc degeneration by not regulating JAK1/STAT3/NLRP3 signal pathway.

Intervertebral disc degeneration (IDD) is a major cause of low back pain. Sinapine thiocyanate (ST) has been reported to have a wide range of biological activities. However, the treatment of IDD with ST has not been studied. To explore the role and mechanism of ST treatment in IDD. Nucleus pulposus cells (NPCs) were induced using lipopolysaccharide (LPS), which was used as an in vitro model of IDD. Cell activity, oxidative stress-related indicators and protein expression were detected using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, enzyme-linked immunosorbent assay (ELISA) and western blot. Pyroptosis was evaluated with propidium iodide (PI)/Hoechst double staining and immunofluorescence for NOD-like receptor protein 3 (NLRP3), and pyroptosis-related proteins and inflammatory factors were measured with western blot and ELISA. The pathological changes of IDD were assessed with hematoxylin & eosin (H&E) and safranin-O staining. Our results showed that ST alleviated LPS-induced degeneration of NPCs, as evidenced by reducing reactive oxygen species (ROS), malondialdehyde (MDA), matrix metalloproteinase-13 (MMP-13), a disintegrin and metalloproteinase with thrombospondin motifs-5 (ADAMTS-5), and increasing collagen II and aggrecan expression. Moreover, ST repressed LPS-induced pyroptosis by inhibiting NLRP3, caspase-1 p20, interleukin (IL)-1β and IL-18. Further studies showed that ST did not restrain the activation of the JAK1/STAT3 signaling pathway induced by colivelin, or of the enhanced pyroptosis induced by polyphyllin VI. Sinapine thiocyanate alleviated IDD in vivo and suppressed NLRP3-mediated pyroptosis and the JAK1/STAT3 signaling pathway. Sinapine thiocyanate could alleviate IDD, although this did not include a reduction in NLRP3-mediated pyroptosis and inactivation of the JAK1/STAT3 signaling pathway, thus potentially being a candidate drug for IDD treatment.

Annulus Fibrosus Injury Induces Acute Neuroinflammation and Chronic Glial Response in Dorsal Root Ganglion and Spinal Cord-An In Vivo Rat Discogenic Pain Model.

Chronic painful intervertebral disc (IVD) degeneration (i.e., discogenic pain) is a major source of global disability needing improved knowledge on multiple-tissue interactions and how they progress in order improve treatment strategies. This study used an in vivo rat annulus fibrosus (AF) injury-driven discogenic pain model to investigate the acute and chronic changes in IVD degeneration and spinal inflammation, as well as sensitization, inflammation, and remodeling in dorsal root ganglion (DRG) and spinal cord (SC) dorsal horn. AF injury induced moderate IVD degeneration with acute and broad spinal inflammation that progressed to DRG to SC changes within days and weeks, respectively. Specifically, AF injury elevated macrophages in the spine (CD68) and DRGs (Iba1) that peaked at 3 days post-injury, and increased microglia (Iba1) in SC that peaked at 2 weeks post-injury. AF injury also triggered glial responses with elevated GFAP in DRGs and SC at least 8 weeks post-injury. Spinal CD68 and SC neuropeptide Substance P both remained elevated at 8 weeks, suggesting that slow and incomplete IVD healing provides a chronic source of inflammation with continued SC sensitization. We conclude that AF injury-driven IVD degeneration induces acute spinal, DRG, and SC inflammatory crosstalk with sustained glial responses in both DRGs and SC, leading to chronic SC sensitization and neural plasticity. The known association of these markers with neuropathic pain suggests that therapeutic strategies for discogenic pain need to target both spinal and nervous systems, with early strategies managing acute inflammatory processes, and late strategies targeting chronic IVD inflammation, SC sensitization, and remodeling.

The relationship between structural changes in paraspinal muscles and intervertebral disc and facet joint degeneration in the lumbar spine of rats

BackgroundDegenerative spine disease is one of the largest causes of disability worldwide and has a multifactorial aetiology. Determining the leading causes of this multifactorial disease could help create new treatment approaches.PurposeStudy the impact of degenerative changes in the paraspinal muscles caused by local (prolonged compression) or systemic (high-fat diet) factors on the structure of the intervertebral discs (IVDs) and facet joints of the lumbar spine in rats.MethodsThe study was conducted using two animal models to create degenerative changes in the paraspinal muscles of 10 white laboratory rats for 90 days and five control rats: 1) high-fat diet model (model 1) involved keeping the rats on a high calorie diet; 2) compression model (model 2) involved binding the paraspinal muscles from L2 to S1 using non-absorbable sutures. Histological analysis for the facet joints and IVDs of rats (at the L1–L4 level) with semi-quantitative analysis of the structure conducted used by degeneration grading system for IVDs and cartilage degeneration score (OARSI) for facet joint.ResultsIn both models, 90 days after the experiment, the degenerative changes observed in the rats’ IVDs were more severe in the annulus fibrosus than in the nucleus pulposus. The height of the IVD in model 1 did not differ from the control group, but in the model 2 was 1.3 times greater (p < 0.001) compared with control. Degenerative changes in the IVD were scored out 5.3 ± 1.7 in model 1 and 5.32 ± 2.1 in model 2 of a possible 16. The height of the articular cartilage of the facet joints was smaller by 1.5 times (p < 0.001) and 1.4 times (p < 0.001) in model 1 and model 2, respectively, compared to the control. Degenerative changes of facet joint were scored out 3.7 ± 0.6 in model 1 and 3.8 ± 0.6 in model 2 of five points according to the cartilage degeneration score.ConclusionsIt was determined that rats who had structural changes in the lumbar paraspinal muscles as a result of being kept on a high-fat diet or subjected to prolonged compression for 90 days, showed degenerative changes in intervertebral discs and osteoarthritis in facet joints of lumbar spine.

An in vivo model of ligamentum flavum hypertrophy from early-stage inflammation to fibrosis.

Multi-joint disease pathologies in the lumbar spine, including ligamentum flavum (LF) hypertrophy and intervertebral disc (IVD) bulging or herniation contribute to lumbar spinal stenosis (LSS), a highly prevalent condition characterized by symptomatic narrowing of the spinal canal. Clinical hypertrophic LF is characterized by a loss of elastic fibers and increase in collagen fibers, resulting in fibrotic thickening and scar formation. In this study, we created an injury model to test the hypothesis that LF needle scrape injury in the rat will result in hypertrophy of the LF characterized by altered tissue geometry, matrix organization, composition and inflammation. An initial pilot study was conducted to evaluate effect of needle size. Results indicate that LF needle scrape injury using a 22G needle produced upregulation of the pro-inflammatory cytokine Il6 at 1 week post injury, and increased expression of Ctgf and Tgfb1 at 8 weeks post injury, along with persistent presence of infiltrating macrophages at 1, 3, and 8 weeks post injury. LF integrity was also altered, evidenced by increases in LF tissue thickness and loss of elastic tissue by 8 weeks post injury. Persistent LF injury also produced multi-joint effects in the lumbar IVD, including disc height loss at the injury and adjacent to injury level, with degenerative IVD changes observed in the adjacent level. These results demonstrate that LF scrape injury in the rat produces structural and molecular features of LF hypertrophy and IVD height and histological changes, dependent on level. This model may be useful for testing of therapeutic interventions for treatment of LSS and IVD degeneration associated with LF hypertrophy.

FOXO3-Activated circFGFBP1 Inhibits Extracellular Matrix Degradation and Nucleus Pulposus Cell Death via miR-9-5p/BMP2 Axis in Intervertebral Disc Degeneration In Vivo and In Vitro

(1) Background: intervertebral disc degeneration (IVDD) defined as the degenerative changes in intervertebral disc is characterized by extracellular matrix (ECM) degradation and death in nucleus pulposus (NP) cells. (2) Methods: The model of IVDD was established in male Sprague Dawley rats using a puncture of a 21-gauge needle at the endplates located in the L4/5 intervertebral disc. Primary NP cells were stimulated by 10 ng/mL IL-1β for 24 h to mimic IVDD impairment in vitro. (3) Results: circFGFBP1 was downregulated in the IVDD samples. circFGFBP1 upregulation inhibited apoptosis and extracellular matrix (ECM) degradation and promoted proliferation in IL-1β-stimulated NP cells. Additionally, circFGFBP1 upregulation mitigated the loss of NP tissue and the destruction of the intervertebral disc structure in vivo during IVDD. FOXO3 could bind to the circFGFBP1 promoter to enhance its expression. circFGFBP1 upregulated BMP2 expression in NP via sponging miR-9-5p. FOXO3 enhanced the protection of circFGFBP1 in IL-1β-stimulated NP cells, whereas a miR-9-5p increase partly reversed the protection. miR-9-5p downregulation contributed to the survival of IL-1β-stimulated NP cells, which was partially reversed by BMP2 silence. (4) Conclusions: FOXO3 could activate the transcription of circFGFBP1 via binding to its promoter, which resulted in the enhancement of BMP2 via sponging miR-9-5p and then inhibited apoptosis and ECM degradation in NP cells during IVDD.

An update on animal models of intervertebral disc degeneration and low back pain: Exploring the potential of artificial intelligence to improve research analysis and development of prospective therapeutics.

Animal models have been invaluable in the identification of molecular events occurring in and contributing to intervertebral disc (IVD) degeneration and important therapeutic targets have been identified. Some outstanding animal models (murine, ovine, chondrodystrophoid canine) have been identified with their own strengths and weaknesses. The llama/alpaca, horse and kangaroo have emerged as new large species for IVD studies, and only time will tell if they will surpass the utility of existing models. The complexity of IVD degeneration poses difficulties in the selection of the most appropriate molecular target of many potential candidates, to focus on in the formulation of strategies to effect disc repair and regeneration. It may well be that many therapeutic objectives should be targeted simultaneously to effect a favorable outcome in human IVD degeneration. Use of animal models in isolation will not allow resolution of this complex issue and a paradigm shift and adoption of new methodologies is required to provide the next step forward in the determination of an effective repairative strategy for the IVD. AI has improved the accuracy and assessment of spinal imaging supporting clinical diagnostics and research efforts to better understand IVD degeneration and its treatment. Implementation of AI in the evaluation of histology data has improved the usefulness of a popular murine IVD model and could also be used in an ovine histopathological grading scheme that has been used to quantify degenerative IVD changes and stem cell mediated regeneration. These models are also attractive candidates for the evaluation of novel anti-oxidant compounds that counter inflammatory conditions in degenerate IVDs and promote IVD regeneration. Some of these compounds also have pain-relieving properties. AI has facilitated development of facial recognition pain assessment in animal IVD models offering the possibility of correlating the potential pain alleviating properties of some of these compounds with IVD regeneration.

The Effect of Diabetes and Disease Duration on Disc Degeneration and ‎Subcutaneous Adipose Tissue Thickness

In this study, we aimed to determine whether the presence and duration of diabetes mellitus (DM) is ‎associated with changes in intervertebral disc degeneration, epiphyseal plaque degeneration, and ‎subcutaneous fat tissue thickness. Diabetic patients and non-diabetic participants (control group) who applied to the ‎internal medicine outpatient clinic between 2020-2022 and had lumbar spine magnetic resonance imaging ‎due to low back and/or leg pain were included the study. Disc degeneration, epiphyseal plaque ‎degeneration, and subcutaneous fat tissue thickness parameters of DM patients and control group were ‎reviewed retrospectively, and the results were statistically evaluated. There were 80 DM patients in the ‎patient group, 25 (31.3%) of these patients were male and 55 (68.8%) were female. There was a total of ‎‎121 participants in the control group, of which 39 (32.2%) were male and 82 (67.8%) were female. The ‎mean age of the patient group was 57.85±10.76 years, while the mean age of the control group was ‎‎54.23±13.09 years. There was no significant difference between the duration of DM and disc degeneration, ‎epiphyseal plaque degeneration, and subcutaneous adipose tissue thickness. However, there was a ‎significant difference between the control group and the DM patient group in terms of disc degeneration and ‎epiphyseal plaque degeneration (p=0.003 and p=0.017, respectively). This study reveals that patients with ‎diabetes have an increased susceptibility to disc degeneration and epiphyseal plaque degeneration, ‎regardless of disease duration. Control and treatment of diabetic disease will make an important contribution ‎to the prevention of secondary complications‎.

Mechanism of Aspirin oxidative stress regulating interleukin-induced apoptosis in nucleus pulposus cells in a rat model of intervertebral disc degeneration.

Intervertebral disc degeneration (IDD) is an important cause of low back pain. Increase of reactive oxygen species (ROS), overexpression of inflammatory factors, and loss of extracellular matrix are important factors in the pathological changes of IDD. The present study aimed to investigate the mechanism of action of Aspirin regulating oxidative stress in IDD, so as to propose new treatment. Nucleus pulposus cells (NPCs) were isolated from the caudal intervertebral discs of Sprague Dawley (SD) rats under sterile conditions. The expression of ROS and inflammatory factors was detected sequentially, and the degree of degeneration of nucleus pulposus cells was observed by real-time fluorescence quantitative polymerase chain reaction (PCR) and cell immunofluorescence staining. In vivo, the caudal disc puncture model was used to induce degeneration, and a local injection of 10 or 100 µg/mL Aspirin was performed. The rats were sacrificed 1 week later, and the disc specimens of the tail vertebrae were collected for imaging, histomorphology, and immunohistochemical analysis. In vitro experiments showed that lipopolysaccharide (LPS) could significantly induce oxidative stress in NPCs and stimulate NPCs to secrete a large amount of ROS and inflammatory factors, which eventually leads to the reduction of collagen type II and polyglycoprotein gene expression in NPCs and the high expression of matrix metalloproteinase (MMP). Consequently, NPCs degeneration occurs. Our results clarified the important role of oxidative stress in IDD and proved that LPS can be used as a drug to alleviate oxidative stress and intervene in the IDD process.

T1ρ, T2 and T2* mapping of lumbar intervertebral disc degeneration: a comparison study

BackgroundEarly and accurate assessment of lumbar intervertebral disc degeneration (IVDD) is very important to therapeutic strategy. This study aims to correlate and compare the performances of T1ρ, T2 and T2* mapping for Pfirrmann grades and morphologic changes in the IVDD.MethodsThis prospective study included 39 subjects with 195 lumbar discs. T1ρ, T2 and T2* mapping were performed, and T1ρ, T2 and T2* values of nucleus pulposus (NP), and anterior and posterior annulus fibrosus were measured. IVDD was assessed with Pfirrmann grading and morphologic changes (normal, bulging, herniation and annular fissure). The performances of T1ρ, T2 and T2* relaxation times were compared for detecting early (Pfirrmann grade II-III) and advanced degeneration (Pfirrmann grade IV–V), as well as for morphologic changes.ResultsT2 relaxation times was strongly corelated with T1ρ and T2* relaxation times. Areas under the curves (AUCs) of T1ρ, T2 and T2* relaxation times of NP were 0.70, 0.87 and 0.80 for early degeneration, and 0.91, 0.95 and 0.82 for advanced degeneration, respectively. AUCs of T1ρ, T2 and T2* relaxation times of NP were 0.78, 0.83 and 0.64 for bulging discs, 0.87, 0.89 and 0.69 for herniated discs, and 0.79, 0.82 and 0.69 for annular tearing, respectively. The AUC of T2 relaxation time was significantly higher than those of T1ρ relaxation times (both P < 0.01) for early IVDD, and the AUCs of T1ρ and T2 relaxation times for assessing advanced degeneration and morphologic changes were similar (P > 0.05) but significantly higher than that of T2*relaxation time (P < 0.01).ConclusionsT2 mapping performed better than T1ρ mapping for the detection of early IVDD. T1ρ and T2 mapping performed similarly but better than T2* mapping for advanced degeneration and morphologic changes of IVDD.

Different phylotypes of Cutibacterium acnes cause different modic changes in intervertebral disc degeneration.

The contribution of Cutibacterium acnes (C. acnes) infection to intervertebral disc degeneration (IDD) and the antibiotic therapy has evoked several controversies in recent years. While some microbiology studies report bacterial disc infection within IDD patients, others attribute the positive results to contamination during prolonged cultures. In addition to the clinical controversy, little was known about the mechanism of C. acnes-caused Modic changes (MCs) if C. acnes was the pathogenic factor. This study aimed to investigate the inflammatory mechanism of MCs induced by different phylotypes of C. acnes in patients with IDD. Specimens from sixty patients undergoing microdiscectomy for disc herniation were included, C. acnes were identified by anaerobic culture, followed by biochemical and PCR-based methods. The identified species of C. acnes were respectively inoculated into the intervertebral discs of rabbits. MRI and histological change were observed. Additionally, we detected MMP expression in the rabbit model using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Of the 60 cases, 18 (30%) specimens were positive for C. acnes, and we identified 4 of 6 defined phylogroups: IA, IB, II and III. The rabbits that received Type IB or II strains of C. acnes showed significantly decreased T1WI and higher T2WI at eighth weeks, while strain III C. acnes resulted in hypointense signals on both T1WI and T2WI. Histological examination results showed that all of the three types of C. acnes could cause disc degeneration and endplates rupture. Moreover, endplate degeneration induced by type IB or II strains of C. acnes is related with MMP13 expression. Meanwhile, strain III C. acnes might upregulated the level of MMP3. This study suggested that C. acnes is widespread in herniated disc tissues. Different types of C. acnes could induce different MCs by increasing MMP expression.

Exercise-induced FNDC5/irisin protects nucleus pulposus cells against senescence and apoptosis by activating autophagy

Intervertebral disc degeneration (IVDD) is a major cause of low back pain (LBP), and excessive senescence and apoptosis of nucleus pulposus (NP) cells are major pathological changes in IVDD. Physical exercise could effectively delay the process of intervertebral disc degeneration; however, its mechanism is still largely unknown. Irisin is an exercise-induced myokine released upon cleavage of the membrane-bound precursor protein fibronectin type III domain-containing protein 5 (FNDC5), and its levels increase after physical exercise. Here, we show that after physical exercise, FNDC5/irisin levels increase in the circulation and NP, senescence and apoptosis are reduced, autophagy is activated in NP tissue, and the progression of IVDD is delayed. Conversely, after knocking out FNDC5, the benefits of physical exercise are compromised. Moreover, the overexpression of FNDC5 in NP tissue effectively alleviated the degeneration of the intervertebral disc (IVD) in rats. By showing that FNDC5/irisin is an important mediator of the beneficial effects of physical exercise in the IVDD model, the study proposes FNDC5/irisin as a novel agent capable of activating autophagy and protecting NP from senescence and apoptosis.

Even mild intervertebral disc degeneration reduces the flexibility of the thoracic spine: an experimental study on 95 human specimens

BACKGROUND CONTEXTIntervertebral disc degeneration represents one of multiple potential trigger factors for reduced passive spinal mobility and back pain. The effects of age-related degenerative intervertebral disc changes on spinal flexibility were however mainly investigated for the lumbar spine in the past, while intervertebral disc degeneration is also highly prevalent in the thoracic spine. PURPOSETo evaluate the effect of the degeneration grade on the range of motion and neutral zone of the thoracic spine. STUDY DESIGNExperimental study including combined radiological grading of intervertebral disc degeneration and biomechanical testing of 95 human thoracic functional spinal units (min. n=4 per level from T1–T2 to T11–T12) from 33 donors (15 female / 18 male, mean age 56 years, age range 37–80 years). METHODSDegeneration grades of the intervertebral discs were assessed using the validated x-ray grading scheme of Liebsch et al. (0=no, 1=mild, 2=moderate, 3=severe degeneration). Motion segments were loaded with pure moments in flexion/extension, lateral bending, and axial rotation to determine range of motion and neutral zone at 5 Nm. RESULTSAll tested specimens exhibited degeneration grades between zero and two. Range of motion significantly decreased for grades one and two compared with grade zero in any motion direction (p<.05), showing the strongest decrease in extension comparing grade two with grade zero (-42%), while no significant differences were detected between grades one and two. Similar trends were found for the neutral zone with the strongest decrease in extension also comparing grade two with grade zero (-47%). Donor age did not significantly affect the range of motion, whereas the range of motion was significantly reduced in specimens from male donors due to the significantly higher degeneration grade in this study. CONCLUSIONSEven mild intervertebral disc degeneration reduces the range of motion and neutral zone of the thoracic spine in any motion plane, whereas progressing degeneration does not further affect its flexibility. This is in contrast to the lumbar spine, where a more gradual decrease of flexibility was found in prior studies, which might be explained by differences between thoracic and lumbar intervertebral disc morphologies. CLINICAL SIGNIFICANCEThoracic intervertebral disc degeneration should be considered as one of multiple potential causal factors in patients showing reduced passive mobility and middle back pain.

Identification of Core Genes and Screening of Potential Targets in Intervertebral Disc Degeneration Using Integrated Bioinformatics Analysis.

Background: Intervertebral disc degeneration (IDD), characterized by diverse pathological changes, causes low back pain (LBP). However, prophylactic and delaying treatments for IDD are limited. The aim of our study was to investigate the gene network and biomarkers of IDD and suggest potential therapeutic targets. Methods: Differentially expressed genes (DEGs) associated with IDD were identified by analyzing the mRNA, miRNA, and lncRNA expression profiles of IDD cases from the Gene Expression Omnibus (GEO). The protein–protein interaction (PPI) network, Gene Ontology (GO) enrichment, Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis as well as miRNA–lncRNA–mRNA networks were conducted. Moreover, we obtained 71 hub genes and performed a comprehensive analysis including GO, KEGG, gene set enrichment analysis (GSEA), gene set variation analysis (GSVA), Disease Ontology (DO), methylation analysis, receiver operating characteristic (ROC) curve analysis, immune infiltration analysis, and potential drug identification. We finally used qRT-PCR to verify 13 significant DEGs in normal and degenerative nucleus pulposus cells (NPCs). Results: We identified 305 DEGs closely related to IDD. The GO and KEGG analyses indicated that changes in IDD are significantly associated with enrichment of the inflammatory and immune response. GSEA analysis suggested that cell activation involved in the inflammatory immune response amide biosynthetic process was the key for the development of IDD. The GSVA suggested that DNA repair, oxidative phosphorylation, peroxisome, IL-6-JAK-STAT3 signaling, and apoptosis were crucial in the development of IDD. Among the 71 hub genes, the methylation levels of 11 genes were increased in IDD. A total of twenty genes showed a high functional similarity and diagnostic value in IDD. The result of the immune cell infiltration analysis indicated that seven genes were closely related to active natural killer cells. The most relevant targeted hub genes for potential drug or molecular compounds were MET and PIK3CD. Also, qRT-PCR results showed that ARHGAP27, C15orf39, DEPDC1, DHRSX, MGAM, SLC11A1, SMC4, and LINC00887 were significantly downregulated in degenerative NPCs; H19, LINC00685, mir-185-5p, and mir-4306 were upregulated in degenerative NPCs; and the expression level of mir-663a did not change significantly in normal and degenerative NPCs. Conclusion: Our findings may provide new insights into the functional characteristics and mechanism of IDD and aid the development of IDD therapeutics.

Treatment of Intervertebral Disc Degeneration

Intervertebral disc degeneration (IDD) causes a variety of signs and symptoms, such as low back pain (LBP), intervertebral disc herniation, and spinal stenosis, which contribute to high social and economic costs. IDD results from many factors, including genetic factors, aging, mechanical injury, malnutrition, and so on. The pathological changes of IDD are mainly composed of the senescence and apoptosis of nucleus pulposus cells (NPCs), the progressive degeneration of extracellular matrix (ECM), the fibrosis of annulus fibrosus (AF), and the inflammatory response. At present, IDD can be treated by conservative treatment and surgical treatment based on patients' symptoms. However, all of these can only release the pain but cannot reverse IDD and reconstruct the mechanical function of the spine. The latest research is moving towards the field of biotherapy. Mesenchymal stem cells (MSCs) are regard as the potential therapy of IDD because of their ability to self‐renew and differentiate into a variety of tissues. Moreover, the non‐coding RNAs (ncRNAs) are found to regulate many vital processes in IDD. There have been many successes in the in vitro and animal studies of using biotherapy to treat IDD, but how to transform the experimental data to real therapy which can apply to humans is still a challenge. This article mainly reviews the treatment strategies and research progress of IDD and indicates that there are many problems that need to be solved if the new biotherapy is to be applied to clinical treatment of IDD. This will provide reference and guidance for clinical treatment and research direction of IDD.

Effect of endplate reduction on endplate healing morphology and intervertebral disc degeneration in patients with thoracolumbar vertebral fracture.

To determine the effect of endplate reduction on the final healing morphology and degenerative changes in intervertebral discs. Forty-eight patients with single-level thoracolumbar fractures with endplate injury were included. All patients underwent posterior reduction and pedicle screw fixation, and postoperative imaging was used to determine whether endplate reduction was successful. The healing morphology of the endplate was divided into three types: increased endplate curvature, irregular healing and traumatic Schmorl node. MRI was performed at baseline and at the last follow-up evaluation to observe changes in disc degeneration (disc height and nucleus pulposus signal) and Modic changes. The reduction rate in the central area was significantly lower than that in the peripheral area (P = 0.017). In patients with successful reduction, 90.9% (20/22) of the endplates healed with increased curvature. In patients with an unsuccessful endplate reduction, 63.4% (26/41) of the endplates healed irregularly, and 34.1% (14/41) of the endplates formed traumatic Schmorl nodes. Endplate reduction was closely related to the final healing morphology of the endplate (P < 0.001), which had a significant protective effect on the degeneration of the intervertebral disc. At the last follow-up evaluation, there was no statistically significant correlation between different endplate healing morphologies and new Modic changes. The reduction rate in the central area is significantly lower than that in the peripheral area. Although all of the intervertebral discs corresponding to fractured endplates had degenerated to different degrees, successful endplate fracture reduction can obviously delay the degeneration of intervertebral discs.

Imaging Evaluation of Intervertebral Disc Degeneration and Painful Discs-Advances and Challenges in Quantitative MRI.

In recent years, various quantitative and functional magnetic resonance imaging (MRI) sequences have been developed and used in clinical practice for the diagnosis of patients with low back pain (LBP). Until now, T2-weighted imaging (T2WI), a visual qualitative evaluation method, has been used to diagnose intervertebral disc (IVD) degeneration. However, this method has limitations in terms of reproducibility and inter-observer agreement. Moreover, T2WI observations do not directly relate with LBP. Therefore, new sequences such as T2 mapping, T1ρ mapping, and MR spectroscopy have been developed as alternative quantitative evaluation methods. These new quantitative MRIs can evaluate the anatomical and physiological changes of IVD degeneration in more detail than conventional T2WI. However, the values obtained from these quantitative MRIs still do not directly correlate with LBP, and there is a need for more widespread use of techniques that are more specific to clinical symptoms such as pain. In this paper, we review the state-of-the-art methodologies and future challenges of quantitative MRI as an imaging diagnostic tool for IVD degeneration and painful discs.

Effects of omega-3 fatty acids and CoQ10 on gross morphology of intervertebral discs subjected to immobilization

The aim of the study was to determine the effects of immobilization on the gross morphology of rats’ intervertebral disc (IVD) and observe the ameliorating effects of Omega 3 fatty acids and Co-enzyme Q 10 (CoQ10). Forty Sprague Dawley rats weighing 250-300 g were procured from NIH Islamabad. The animals were randomly selected and were divided into four groups of 10 animals in each. Group-A rats served as control group. Each rat of Group B was disc immobilized by using an Illizarov-type apparatus, which was applied for 60 days. Group-C and -D rats after disc immobilization were administrated with Omega 3 fatty acids (260 mg/kg/day) and CoQ10 (150mg/kg/day) through oral gavage respectively. Gross examination of IVD was done using the Thompson grading scale and the disc alterations were scored from grade 1 to 5 in increasing order of IVD alterations. Gross examination of the sections of IVD’s of the control group showed normal healthy morphology, falling in Thompson grade I degeneration. The frequency of disc alteration was statistically significant in disc-immobilized group B when compared to control group A (p-value=0.000), group C (p-value=0.000) and group D (p value=0.002). Group C in which n-3 fatty acid was given along with disc immobilization, showed significant improvement in disc degenerative changes. On comparison with group B, p-value&lt;0.001 was statistically significant. In experimental Group D, where CoQ10 was given along with disc immobilization, the degenerative changes were significantly reduced as compared to Group B (p = 0.002). In this study, gross morphological changes were induced by immobilization in IVDs of the experimental rats and its reversal by omega 3 and CoQ10 was proven. Co-administration of Omega 3 and CoQ10 significantly minimized degenerative changes in IVDs induced by immobilization.

AMPK as a Potential Therapeutic Target for Intervertebral Disc Degeneration.

As the principal reason for low back pain, intervertebral disc degeneration (IDD) affects the health of people around the world regardless of race or region. Degenerative discs display a series of characteristic pathological changes, including cell apoptosis, senescence, remodeling of extracellular matrix, oxidative stress and inflammatory local microenvironment. As a serine/threonine-protein kinase in eukaryocytes, AMP-activated protein kinase (AMPK) is involved in various cellular processes through the modulation of cell metabolism and energy balance. Recent studies have shown the abnormal activity of AMPK in degenerative disc cells. Besides, AMPK regulates multiple crucial biological behaviors in IDD. In this review, we summarize the pathophysiologic changes of IDD and activation process of AMPK. We also attempt to generalize the role of AMPK in the pathogenesis of IDD. Moreover, therapies targeting AMPK in alleviating IDD are analyzed, for better insight into the potential of AMPK as a therapeutic target.