Pathogenesis Of Lumbar Disc Herniation Research Articles

Failure mechanical properties of lumbar intervertebral disc under high loading rate

BackgroundLumbar disc herniation (LDH) is the main clinical cause of low back pain. The pathogenesis of lumbar disc herniation is still uncertain, while it is often accompanied by disc rupture. In order to explore relationship between loading rate and failure mechanics that may lead to lumbar disc herniation, the failure mechanical properties of the intervertebral disc under high rates of loading were analyzed.MethodBend the lumbar motion segment of a healthy sheep by 5° and compress it to the ultimate strength point at a strain rate of 0.008/s, making a damaged sample. Within the normal strain range, the sample is subjected to quasi-static loading and high loading rate at different strain rates.ResultsFor healthy samples, the stress–strain curve appears collapsed only at high rates of compression; for damaged samples, the stress–strain curves collapse both at quasi-static and high-rate compression. For damaged samples, the strengthening stage becomes significantly shorter as the strain rate increases, indicating that its ability to prevent the destruction is significantly reduced. For damaged intervertebral disc, when subjected to quasi-static or high rates loading until failure, the phenomenon of nucleus pulposus (NP) prolapse occurs, indicating the occurrence of herniation. When subjected to quasi-static loading, the AF moves away from the NP, and inner AF has the greatest displacement; when subjected to high rates loading, the AF moves closer to the NP, and outer AF has the greatest displacement. The Zhu–Wang–Tang (ZWT) nonlinear viscoelastic constitutive model was used to describe the mechanical behavior of the intervertebral disc, and the fitting results were in good agreement with the experimental curve.ConclusionExperimental results show that, both damage and strain rate have a significant effect on the mechanical behavior of the disc fracture. The research work in this article has important theoretical guiding significance for preventing LDH in daily life.

Protectin D1 ameliorates non-compressive lumbar disc herniation through SIRT1-mediated CGRP signaling.

Background. Neuro-inflammatory response promotes the initiation and sustenance of lumbar disc herniation (LDH). Protectin D1 (PD1), as a new type of specialized pro-resolving mediator (SPM), can improve the prognosis of various inflammatory diseases. Recent studies have shown that over representation of calcitonin gene-related peptides (CGRP) may activate nociceptive signaling following nerve injury. Silent information regulator 1 (SIRT1) is ubiquitously expressed in the dorsal horn of the spinal cord and plays a role in the pathogenesis of LDH. In this study, we investigated the analgesic effects of PD1 and elucidated the impact of neurogenic inflammation in the pathogenesis of neuropathic pain induced by non-compressive lumbar disc herniation (NCLDH) in a rat model. Methods. NCLDH models were established by applying protruding autologous nucleus pulposus to the L5 Dorsal root ganglion (DRG). PD1, SIRT1 antagonist or agonist, CGRP or antagonist were administered as daily intrathecal injections for three consecutive days postoperatively. Behavioral tests were conducted to assess mechanical and thermal hyperalgesia. The ipsilateral lumbar (L4-6) segment of the spinal dorsal horn was isolated for further analysis. Alterations in the release of SIRT1 and CGRP were explored using western blot and immunofluorescence. Results. Application of protruded nucleus (NP) materials to the DRG induced mechanical and thermal allodynia symptoms, and deregulated the expression of pro-inflammatory and anti-inflammatory cytokines in rats. Intrathecal delivery of PD1 significantly reversed the NCLDH-induced imbalance in neuro-inflammatory response and alleviated the symptoms of mechanical and thermal hyperalgesia. In addition, NP application to the DGRs resulted the spinal upregulation of CGRP and SIRT1 expression, which was almost restored by intrathecal injection of PD1 in a dose-dependent manner. SIRT1 antagonist or agonist and CGRP or antagonist treatment further confirmed the result. Conclusion. Our findings indicate PD1 has a potent analgesic effect, and can modulate neuro-inflammation by regulating SIRT1-mediated CGRP signaling in NCLDH.

Integrated analysis of single-cell and bulk RNA sequencing data identifies the characteristics of ferroptosis in lumbar disc herniation.

Lumbar disc herniation (LDH) is the most common condition associated with low back pain, and it adversely impacts individuals' health. Ferroptosis has recently emerged as a novel factor in the pathogenesis of LDH; however, the specific impacts of ferroptosis on LDH have not been fully elucidated. Ferroptosis-related differentially expressed genes (FRDEGs) were identified from the transcriptomic datasets of LDH. Gene set enrichment analysis (GSEA) was conducted to identify biological mechanism and related pathways. LASSO and SVM-RFE algorithms were applied to detect signature genes. Function of the signature gene was confirmed by RT-qPCR. The CIBERSORT algorithm was used to compare immune infiltration between LDH and normal samples. Correlation analysis between MYB and immune cells was analyzed using the Pearson method. Additionally, we used scRNA-seq to dissect cell clusters and cellular interactions. AUCell scoring was used to analyze the ferroptosis scores of different cell types. We found that MYB, a highly expressed ferroptosis-related gene, was associated with LDH By leveraging bioinformatics analysis. In immune infiltration analysis, the abundance of monocytes and macrophages varied significantly between the LDH group and disc spondylolisthesis (DS) group. MYB was correlated with most immune cells. GSEA revealed MYB was significantly enriched in immune-related pathways. Furthermore, scRNA-seq analysis revealed the presence of eight distinct cell types. AUCell analysis showed that macrophages had a high ferroptosis score. Cell trajectory analysis revealed that chondrocyte 1 was at the beginning of the trajectory, while calcification inhibiting chondrocytes and fibrochondrocytes accumulated along the middle and tail end of the trajectory, respectively. Cell-cell communication analysis identified chondrocyte 1 had an extensive communication network with other clusters and interacted with nucleus pulposus through collagen signaling pathway. Our analysis demonstrated that MYB may be a potential therapeutic target for LDH. This study provides a resource for the orthopedics community that will facilitate additional discoveries directedly toward understanding the pathogenesis process of LDH.

Posterior longitudinal ligament suturation after lumbar discectomy provides postoperative a large intradural area: First report.

Stability and flexibility of the spine are provided by the posterior longitudinal ligament (PLL). It plays a key role in the pathogenesis of lumbar disc herniation (LDH) by preventing disc protrusion. The effect of the suturing of the PLL on the intradural area was investigated. The patients were included in whom lumbar microdiscectomy was performed between January 2021 and July 1, 2022. The patients were randomly divided into two groups as PLLs were sutured and unsutured. Forty-six (23 males and 23 females) patients were included. The PLLs were sutured in 22 patients (Group 1) and not sutured in 24 patients (Group 2). The levels, sides of LDHs, and ages and gender of patients were also analyzed in both groups, which were not statistically significant. Preoperative mean spinal intradural areas were 77.29 mm2 for the PLL unsutured group and 85.40 mm2 for the PLL sutured group (Groups 1 and 2). For patients in Groups 1 and 2, the postoperative mean spinal intradural areas grew to 134.73 mm2 and 96.12 mm2, respectively. The difference in preoperative mean spinal intradural regions between the two groups was not statistically significant; however, Group 1 showed a substantial difference (sutured PLL patients). This study first time indicates that suturing PLL has a protective and supportive role in patients who were operated on for LDH.

Application of Muscle and Bone Balance Theory in Lumbar Disc Herniation

Pingle Guo's theory of "Balance of Muscles and Bones" has a long history. Muscles and bones are fundamental to each other to maintain the static and dynamic balance of the spine and ensure the normal movement of the spine joint. Muscle and bone balance is the normal physiological state of lumbar vertebrae, and the imbalance of muscle and bone is the basic pathogenesis of lumbar disc herniation, which is caused by "slot out of muscle" and "bone dislocation". It is an important principle to treat prolapse of lumbar intervertebral disc by massage. Aiming at the tenderness and percussion pain of soft tissue around the waist and the limitation of muscular activity around the waist, and the "osseosynostosis" of the lumbar vertebrae, which is mainly manifested by the reduction or disappearance of the physiological protruding of the lumbar vertebrae, or the decrease of the lumbar vertebrae's activity, or the different degree of scoliosis, etc. Therefore, the application of "muscle and bone balance" theory of clinical diagnosis and treatment of lumbar disc herniation is of great significance.

Role of the Lumbosacral Transition Vertebra and Vertebral Lamina in the Pathogenesis of Lumbar Disc Herniation.

ObjectiveTo investigate the prevalence of lumbosacral transition vertebrae (LSTVs) in both the normal population and the lumbar disc herniation (LDH) population and to determine the risk factors for LDH.MethodsBetween January 2019 and September 2020, all patients aged 18–39 years and underwent an anteroposterior (AP) X‐ray of the lumbar vertebrae were retrospective reviewed in our institution. Those patients who were diagnosed with LDH were eligible for inclusion in the LDH group. During the same period, those patients admitted to our hospital who underwent an anteroposterior X‐ray of the lumbar spine and had not been diagnosed with LDH were included in the control group. Those patients with disease that might affect the lumbar anatomy were excluded from both groups. The type of LSTV was classified according to the Castellvi classification. The height of the lumbar vertebral lamina was evaluated through the h/H index. The inter‐ and intra‐observer reliability was evaluated by one senior radiologist and one senior orthopedist using intraclass correlation coefficient (ICC). The association between the LSTV and the herniation level was also investigated. Binary logistic regression was used to explore the association of different factors between the LDH group and the control group.ResultsTwo hundred LDH patients (115 male and 85 female) and 200 individuals (108 male and 92 female) were investigated retrospectively. The prevalence of LSTVs was 71.5% (n = 143) in the LDH group and 34.0% (n = 68) in the control group. The most frequent LSTV types were type Ib and type IIa. The inter‐ and intra‐observer ICCs of the measurement of “h/H” index and the classification of LSTV were all “excellent” (ICC > 0.90). The median h/H index in the control group was significantly higher than that in the LDH group (0.28 (0.26, 0.31) vs 0.34 (0.31, 0.37), P = 0.000). The distribution of the Castellvi classification in the L4/5 and L5/S1 herniation patients was significantly different (P = 0.048). LSTVs, BMI and the h/H index were closely associated with LDH, with odds ratios of 3.06 (95% CI: 2.12–4.43), 1.23 (95% CI: 1.13–1.33) and 0.09 (95% CI: 0.05–0.15), respectively. The incidence of L4/5 disc herniation in patients with an LSTV was significantly more common than that in patients with L5/S1 disc herniation (P = 0.048).ConclusionThe prevalence of LSTVs was 34.0% in the control group and 71.5% in the LDH group; LSTVs and BMI were positively correlated with LDH, and h/H was negatively correlated with LDH.

Comparison of the histopathological differences between the spinal material and posterior longitudinal ligament in patients with lumbar disc herniation: A focus on the etiopathogenesis.

BACKGROUND: Lumbar disc herniation (LDH) occurs owing to the inability of the posterior longitudinal ligament (PLL) to preserve the disc material within the intervertebral space. There is apparently no study that has investigated the histopathological changes occurring in both PLL and disc material in patients with LDH.OBJECTIVE: Investigate and compare the histopathological changes occurring in PLL and disc material of the patients who underwent a surgical operation for LDH.DESIGN: Descriptive, cross-sectional.SETTING: Pathology and neurosurgery departments of a tertiary health care institutionPATIENTS AND METHODS: The study included patients who underwent surgical operation for LDH from January 2018 to May 2019 and whose PLL and disc material were removed together, and had disc degeneration findings that were radiologically and histologically concordant.MAIN OUTCOME MEASURES: PLL degeneration scores according to the histopathological findings, changes in disc materials according to the MRI findings, disc degeneration scores according to the histo-pathological findings.SAMPLE SIZE: 50.RESULTS: MRI and histological examinations showed fully degenerated black discs (Grade 2) in 12 patients, partially degenerated discs (Grade 1) in 29 patients and fresh/acute discs (Grade 0) in 9 patients. The PLL showed grade 0 degeneration in 2 patients, grade 1 degeneration in 23 patients, and grade 2 degeneration in 25 patients. PLL degeneration grades were higher than the disc degeneration grades (P=.002).CONCLUSION: Longitudinal ligament degeneration can play a significant role in the pathogenesis of LDH. To the best of our knowledge, this study represents the first to focus on the histopathological changes occurring in both the PLL and disc material in patients with LDH.LIMITATIONS: Small sample, retrospectiveCONFLICT OF INTEREST: None.

RAB40C gene polymorphisms rs62030917 and rs2269556 are associated with an increased risk of lumbar disc herniation development in the Chinese Han population.

Lumbar disc herniation (LDH) places a serious burden on the daily lives and socioeconomics of people. Although the pathogenesis of LDH is complex, genetic factors such as single nucleotide polymorphisms (SNPs) may affect the risk of developing LDH. In the present study, we aimed to elucidate the effect of RAB40C SNPs on the risk of LDH in the Chinese Han population. We investigated 508 LDH cases and 508 healthy controls for this case-control study. Three tag SNPs in RAB40C were selected and genotyped using the Agena MassARRAY platform (Agena Bioscience, San Diego, CA, USA). After adjusting for age and gender, odds ratios (ORs) and 95% confidence intervals (CIs) were calculated using unconditional logistic regression. In the allele model, we found rs62030917 and rs2269556 in RAB40C with a minor G allele significantly increased the risk of LDH (rs62030917: OR = 1.23, 95% CI = 1.00-1.50, p = 0.046; rs2269556: OR = 1.21, 95% CI = 1.02-1.45, p = 0.033). In genetic model analysis, rs2269556 was associated with an increased risk of LDH under both codominant (OR = 1.49, 95% CI = 1.03-2.15, p = 0.035) and log-additive models (OR = 1.21, 95% CI = 1.01-1.45, p = 0.035). rs62030917 of RAB40C was associated with an increased risk of LDH under codominant, recessive and log-additive models (p < 0.05) only among individuals younger than 49 years after stratification by age. For the first time, our results suggest that rs62030917 and rs2269556 in the RAB40C gene influence genetic susceptibility to LDH.

The impact of spino-pelvic parameters on pathogenesis of lumbar disc herniation.

Controversies exist on the relative contribution of sagittal spino-pelvic alignment on pathogenesis of lumbar disc herniation. Spinopelvic alignment parameters, pelvic incidence, pelvic tilt, sacral slope, pelvic radius-S1 end-plate ratio and pelvic radius-S1 angle, in 52 patients with lumbar disc herniation were compared with the same measurements in 43 healthy adult volunteers. Statistical analysis revealed significant difference in pelvic incidence and sacral slope between patients with lumbar disc herniation and normal population. Patients with lumbar disc herniation have a more vertical sacrum compared to the normal population which may result in higher intradiscal pressure in these patients.

A double-network hydrogel for the dynamic compression of the lumbar nerve root.

Current animal models of nerve root compression due to lumbar disc herniation only assess the mechanical compression of nerve roots and the inflammatory response. Moreover, the pressure applied in these models is static, meaning that the nerve root cannot be dynamically compressed. This is very different from the pathogenesis of lumbar disc herniation. In this study, a chitosan/polyacrylamide double-network hydrogel was prepared by a simple two-step method. The swelling ratio of the double-network hydrogel increased with prolonged time, reaching 140. The compressive strength and compressive modulus of the hydrogel reached 53.6 and 0.34 MPa, respectively. Scanning electron microscopy revealed the hydrogel's crosslinked structure with many interconnecting pores. An MTT assay demonstrated that the number of viable cells in contact with the hydrogel extracts did not significantly change relative to the control surface. Thus, the hydrogel had good biocompatibility. Finally, the double-network hydrogel was used to compress the L4 nerve root of male sand rats to simulate lumbar disc herniation nerve root compression. The hydrogel remained in its original position after compression, and swelled with increasing time. Edema appeared around the nerve root and disappeared 3 weeks after operation. This chitosan/polyacrylamide double-network hydrogel has potential as a new implant material for animal models of lumbar nerve root compression. All animal experiments were approved by the Animal Ethics Committee of Neurosurgical Institute of Beijing, Capital Medical University, China (approval No. 201601006) on July 29, 2016.

Evaluation of Common Variants in Matrix Metalloproteinase-9 Gene with Lumbar Disc Herniation in Han Chinese Population.

Lumbar disc herniation (LDH) is a common and frequent orthopedic disease with strong genetic determinants. The disruption of the intervertebral disc extracellular matrix has been found to play a key role in the development of LDH, suggesting that abnormal matrix metalloproteinases (MMPs) may promote the degradation of the disc matrix. MMP-9, an important member of the MMP family, is a good candidate for the LDH susceptibility gene. The present study aimed to investigate the association of common variants in the MMP-9 gene with the risk, severity, and clinical characteristic variables of LDH. Fourteen tag single nucleotide polymorphisms (SNPs) entirely covering the region of the MMP-9 gene were analyzed in a sample of 845 patients and 1751 healthy controls. The SNP rs17576 was found to be significantly associated with susceptibility to LDH (OR = 0.77, p = 0.0002), which was also confirmed by haplotype-based analyses (rs79845319-rs17576-rs45437897, global p < 0.001). Our results indicated that the A allele of rs17576 reduced the risk of LDH by ∼23% on average. Furthermore, the G allele of rs17576 was found to correlate with more severe grades of disc degeneration. Our results provide additional evidence supporting an important role of the MMP-9 gene in the pathogenesis of LDH.

Analysis of Spino-pelvic Sagittal Alignment in Young Chinese Patients with Lumbar Disc Herniation.

Few studies have concentrated on the sagittal alignment of lumbar disc herniation (LDH), especially the parameters of the pelvis, and controversy exists about whether pelvic morphology is involved in the pathogenesis of LDH. The present study analyzed the characteristics of the sagittal alignment in young Chinese LDH patients and explored the impact of pelvic morphology on the pathogenesis of LDH. A retrospective analysis was conducted on 100 young patients with LDH (69 men and 31 women, aged 18-35 years), and the clinical and imaging findings met the criteria for the diagnosis of LDH. The control group included 100 asymptomatic volunteers with matching age and gender. Coronal and sagittal parameters were measured on the anteroposterior and lateral radiographs of the whole spine, including lumbar lordosis (LL), pelvic incidence (PI), sacral slope (SS), pelvic tilt (PT), thoracic kyphosis (TK), and sagittal balance (SVA). The cases were classified into four types by the apex position of lumbar lordosis (type I, L5 or the L4-5 intervertebral space; type II, bottom or middle of L4 ; type III, upper part of L4 or the intervertebral space between L3 and L4 ; type IV, L3 or more high level), and divided into three groups by PI; namely, a low PI group (PI < 40°), a medium PI group (40° ≤ PI < 50°), and a high PI group (PI ≥ 50°). The sagittal parameters, especially PI, were compared between the LDH group and the control group. Correlations between the parameters in the LDH group were analyzed. The PI value of the LDH group was not different from that of the control group (46.1° ± 10.0° vs 47.2° ± 8.8°, P > 0.05). The LDH group showed lower average LL, SS, and TK (P < 0.01), as well as higher PT and SVA compared with the control group (P < 0.01). The LL (34.4° ± 15.3° vs 50.8° ± 10.2°) and SVA (21.6 ± 53.6 mm vs - 18.4 ± 32.8 mm) showed a significant difference (P < 0.01); LL was correlated with PI, SS, PT, TK and SVA (P < 0.01); and SVA was also correlated with the parameters above (P < 0.01) except PI (P > 0.05), and the lordosis apex tended to be higher. The distributions of PI groups between the LDH group and the control group were not different. Pairwise correlations were found among LL, PI, SS, and PT. In addition, TK and SVA were correlated with LL, SS, and PT. There is no difference in PI between young Chinese patients with lumbar disc herniation and the normal population. Young LDH patients may present flat lumbar and thoracic curves, as well as lower sacral slope. The anteversion sagittal imbalance is regulated by both the spine and the pelvis.

Interleukin-23 may contribute to the pathogenesis of lumbar disc herniation through the IL-23/IL-17 pathway.

BackgroundStudies have indicated that interleukin 23 (IL-23) plays an important role in many inflammatory- and autoimmune-related diseases. However, there is little knowledge about IL-23 in lumbar disc herniation (LDH). Thus, in this study, we aimed to find out whether IL-23 is expressed in intervertebral discs (IVDs) and what roles it may play.MethodsHuman IVD tissues were collected from 29 LDH patients and 8 vertebral fracture patients (normal control, NC group). According to the integrity of annulus fibrosus, LDH patients were divided into two groups: R group (ruptured group, n = 16) and NR group (non-ruptured group, n = 13). Morphological changes of IVDs were assessed by hematoxylin and eosin (HE staining), and expression of IL-23 in IVD tissues was detected by immunohistochemical staining. Besides gene expression of IL-23, IL-17, IL-6, IL-1β, and TNF-α was also evaluated by reverse transcription polymerase chain reaction (RT-PCR).ResultsThe results showed that the R group was more degenerated than the other two groups and NC group showed the least degenerated performance; stronger positive IL-23 expression was observed in herniated IVDs, especially in the R group. Meanwhile, higher gene expression of IL-23, IL-17, IL-6, IL-1β, and TNF-α was found in the tissues from LDH patients and a positive correlation between IL-17 and IL-23 gene expression was also observed.ConclusionsTaken all above results together, it may be deduced that higher expression of IL-23 may contribute to the deterioration of IVDs through the IL-23/IL-17 pathway.

Role of interleukin-17 in chondrocytes of herniated intervertebral lumbar discs.

Lumbar disc herniation (LDH) is a common cause of lumbosacral radiculopathy. An autoimmune response to a herniated nucleus pulposus (NP) has been suggested to play an important role in the initiation of radiculopathy. Interleukin-17 (IL-17) is a cytokine associated with inflammation and autoimmunity. The presence of IL-17 has been studied in patients with LDH; however, extensive investigation into the expression of IL-17 in different disc pathologies of LDH has not yet been conducted. The aim of the present study was to investigate the role of neovascularization and hypertrophic chondrocytes in herniated intervertebral lumbar discs. Fifty-two intervertebral lumbar disc specimens were extracted from 46 patients with LDH and were subsequently classified as either contained or non-contained disc herniation (CDH and NCDH, respectively). The specimens were stained with hematoxylin and eosin or toluidine blue, or were immunostained with polyclonal antibodies to IL-17 using the streptavidin-peroxidase method. The neovascular tissue and staining results were graded to establish the histological differences between the two herniation types. The intervertebral discs (IVDs) obtained from patients with NCDH showed significantly more neovascularization and granulation tissue than the discs obtained from patients with CDH (P<0.05). Furthermore, hypertrophic chondrocytes were more abundant in the NCDH specimens than in the CDH specimens (P<0.05). Similarly, the number of IL-17-immunoreactive cells was significantly higher in the NCDH specimens than that in the CDH specimens (P<0.01). In conclusion, local inflammation and autoreactive immune activation may play an important role in the pathogenesis of LDH. These results also suggest a role of chondrocytes in the repair of herniated IVDs.

Akt/PKB isoforms expression in the human lumbar herniated disc: correlation with clinical and MRI findings

Intervertebral disc (IVD) degeneration suggests a complex process influenced by genetics, lifestyle and biomechanics, which accounts for the development of low back pain (LBP) and lumbar radiculopathy, a major cause of musculoskeletal disability in humans. The family of Akt/PKB kinases is a principal mediator in the signal transduction pathways, which contribute to transcriptional regulation, cell growth, proliferation, apoptosis, and survival ability. The purpose of this study was to evaluate the transcriptional profile of the AKT family genes in human herniated discs and the involvement of the PI3K-Akt signaling pathway in human IVD degeneration. Real-time PCR analysis was used to assess the mRNA expression pattern of the three Akt/PKB isoforms in 63 herniated and 10 control disc specimens. Our results showed a significant positive correlation between AKT1 and AKT3 mRNA in herniated discs suggesting a synergistic action between these isoforms in disc herniation. Interestingly, AKT2 mRNA was up-regulated in patients with acute pain during the first 12 months, indicating that AKT2 transcriptional activation may be associated with acute rather than chronic inflammation and phagocytosis. Finally, Akt1/PKB transcription presented a stepwise activation as disc herniation deteriorated. Our findings provide evidence on the transcriptional activation of the Akt/PKB pathway indicating that it is involved in lumbar disc degeneration. There is need for further studies to elucidate the exact role and down-stream signaling action of Akt/PKB isoforms in the pathogenesis of lumbar disc herniation.

Correlation of matrix metalloproteinases–1 and –3 with patient age and grade of lumbar disc herniation

The authors studied the histological alterations and the expression of matrix metalloproteinase (MMP)-1 and MMP-3 in disc specimens of patients who had undergone operations for lumbar disc herniation. Forty-three lumbar disc specimens were evaluated histopathologically for degenerative changes and immunohistochemical expression of MMP-1 and MMP-3. The observed degenerative changes provided a degenerative score that was applied in each patient. Sections of disc immunostained for MMP-1 and MMP-3 were evaluated semiquantitatively. Patients were categorized in 3 age groups: < 30 years, between 30 and 60 years, and > 60 years of age. The expression of MMP-1 and MMP-3 were correlated to patient's age, degenerative score, and grade of lumbar disc herniation. There was no statistically significant difference in the degenerative score between the age groups. Degenerative changes were more pronounced in greater grades of herniation (p < 0.0001). In the group of patients < 30 years of age there was a significant correlation between MMP-1 and MMP-3 expression and both degenerative score and herniation grade. For the group of patients 30-60 years of age, there was no significant difference between MMP-1 expression and degenerative score, but the correlation between MMP-1 expression and grade of herniation was significant. There was a significant correlation between MMP-3 expression and both degenerative score and herniation grade. Regarding the patients > 60 years of age, there was a significant correlation between MMP-1 and MMP-3 expression and both degenerative score and herniation grade. There was a significantly lower expression of both MMP-1 and MMP-3 in the group < 30 years of age compared with the other ages. No significant correlation was found in MMP-1 and MMP-3 expression between the groups of patients who were 30-60 and > 60 years of age. Interestingly, in age groups > 30 years, there were no statistically significant differences between the expression of MMP-1 and MMP-3, whereas in patients < 30 years of age the expression of MMP-3 was significantly lower than the expression of MMP-1. The expression of MMP-1 and MMP-3 were strongly correlated to the age of the patients and the grade of herniation. An important finding in this study is the differential expression of MMP-1 and MMP-3 between the age groups. In the young age group it appears that deregulation of MMP-1 expression is higher than that of MMP-3 in the pathogenesis of lumbar disc herniation.

A Functional Polymorphism in THBS2 that Affects Alternative Splicing and MMP Binding Is Associated with Lumbar-Disc Herniation

Lumbar-disc herniation (LDH), one of the most common musculoskeletal diseases, has strong genetic determinants. Recently, several genes that encode extracellular matrix (ECM) proteins in the intervertebral disc have been reported to associate with LDH. Thrombospondins (THBSs) 1 and 2 are good candidates for the LDH susceptibility gene: They are intervertebral disc ECM proteins that regulate the effective levels of matrix metalloproteinases (MMPs) 2 and 9, which are key effectors of ECM remodeling. Here, we report that THBS2 is associated with LDH in Japanese populations. An intronic SNP in THBS2 (IVS10-8C --> T; rs9406328) showed significant association (p = 0.0000028) with LDH in two independent Japanese populations. This SNP, located in a polypyrimidine tract upstream of the 3' splice site of intron 10, exerts allelic differences on exon 11 skipping rates in vivo, with the susceptibility allele showing increased skipping. Skipping of exon 11 results in decreased THBS2 interaction with MMP2 and MMP9. Further, a missense SNP in MMP9 (Q279R; rs17576) is also strongly associated with LDH in the Japanese population (p = 0.00049) and shows a combinatorial effect with THBS2 (odds ratio 3.03, 95% confidence interval 1.58-5.77). Thus, a splicing-affecting SNP in THBS2 and a missense SNP in MMP9 are associated with susceptibility to LDH. Our data indicate that regulation of intervertebral disc ECM metabolism by the THBS2-MMP system plays an essential role in the etiology and pathogenesis of LDH.

The role of cyclooxygenase-2 in lumbar disc herniation.

The expression of cyclooxygenase-2 was studied immunohistologically in specimens from lumbar disc herniation. The cultured disc cells also were examined to evaluate the significance of cyclooxygenase-2, which might be involved in the pathogenesis of lumbar disc herniation. To investigate whether cyclooxygenase-2 might be involved in the pathogenesis of lumbar disc herniation. Prostaglandin E2 is one of the most important mediators contributing to pathogenetic components of lumbar disc herniation. Cyclooxygenase-2, the rate-limiting enzyme of prostaglandin E2 synthesis, has been identified and extensively investigated in other inflammatory diseases. However, the role of cyclooxygenase-2 in lumbar disc herniation has never been addressed. Fifteen specimens from patients with lumbar disc herniation and five control discs from traumatic burst fracture were harvested. The expression of cyclooxygenase-2 was evaluated immunohistologically. The ability of cultured disc cells to produce prostaglandin E2 with inflammatory stimulus in the presence or absence of a selective inhibitor of cyclooxygenase-2 was investigated. At the same time, the induction of cyclooxygenase-2 mRNA of these cells by reverse transcriptase-polymerase chain reaction was detected. The manner in which this prostaglandin E2 production could be suppressed by various doses of a cyclooxygenase-2 inhibitor also was investigated. Immunohistologically, the expression of cyclooxygenase-2 was observed only in the lumbar disc herniation specimens. The cultured cells had a strong ability to produce prostaglandin E2 coinciding with cyclooxygenase-2 mRNA induction. A selective inhibitor of cyclooxygenase-2 inhibited this prostaglandin E2 production in a dose-dependent manner. Cyclooxygenase-2 might be involved in the pathogenesis of lumbar disc herniation through upregulation of prostaglandin E2 production.