Paraspinal muscle gene expression across different aetiologies in individuals undergoing surgery for lumbar spine pathology.

PURPOSE: The purpose of this study was to understand potential baseline transcriptional expression differences in paraspinal skeletal muscle from patients with different underlying lumbar pathologies by comparing multifidus gene expression profiles across individuals with either disc herniation, facet arthropathy, or degenerative spondylolisthesis. METHODS: Multifidus biopsies were obtained from patients (n = 44) undergoing lumbar surgery for either disc herniation, facet arthropathy, or degenerative spondylolisthesis. Diagnostic categories were based on magnetic resonance images, radiology reports, and intraoperative reports. Gene expression for 42 genes was analysed using qPCR. A one-way analysis of variance was performed for each gene to determine differences in expression across diagnostic groups. Corrections for multiple comparisons across genes (Benjamini-Hochberg) and for between-group post hoc comparisons (Sidak) were applied. RESULTS: Adipogenic gene (ADIPOQ) expression was higher in the disc herniation group when compared to the facet arthropathy group (p = 0.032). Adipogenic gene (PPARD) expression was higher in the degenerative spondylolisthesis group when compared to the disc herniation group (p = 0.013), although absolute gene expression levels for all groups was low. Fibrogenic gene (COL3A1) had significantly higher expression in the disc herniation group and facet arthropathy group when compared to the degenerative spondylolisthesis group (p < 0.001 and p = 0.038, respectively). When adjusted for multiple comparisons, only COL3A1 remained significant (p = 0.012). CONCLUSION: Individuals with disc herniation and facet arthropathy demonstrate higher COL3A1 gene expression compared to those with degenerative spondylolisthesis. Future research is required to further understand the biological relevance of these transcriptional differences.

Lumbar intervertebral disc mRNA sequencing identifies the regulatory pathway in patients with disc herniation and spondylolisthesis.

Lumbar degenerative disc disease (DDD) is a multifaceted progressive condition and often accompanied by disc herniation (DH) and/or degenerative spondylolisthesis (DS). Given the high prevalence of the disease (up to 20% according to some estimates) and the high costs associated with its care, there is a need to explore novel therapies such as regenerative medicine. Exploring these novel therapies first warrants investigation of molecular pathways underlying these disorders. Here, we show results from next generation RNA sequencing (RNA-seq) on mRNA isolated from 10 human nucleus pulposus (NP) samples of lumbar degenerated discs (DH and DS; n = 5 for each tissue) and other musculoskeletal tissues (Bone, cartilage, growth plate, and muscle; n = 7 for each tissue). Pathway and network analyses based on gene ontology (GO) terms were used to identify the biological functions of differentially expressed mRNAs. A total of 701 genes were found to be significantly upregulated in lumbar NP tissue compared to other musculoskeletal tissues. These differentially expressed mRNAs were primarily involved in DNA damage, immunity and G1/S transition of mitotic cell cycle. Interestingly, DH-specific signaling genes showed major network in chemotactic (e.g., CXCL10, CXCL11, IL1RL2 and IL6) and matrix-degrading pathway (e.g., MMP16, ADAMTSL1, 5, 8, 12, and 15), while DS-specific signaling genes were found to be those involved in cell adhesion (e.g., CDH1, EPHA1 and EFNA2) and inflammatory cytokines (e.g., CD19, CXCL5, CCL24, 25 and XCL2). Our findings provide new leads for therapeutic drug discovery that would permit optimization of medical or pharmacological intervention for cases of lumbar DDD.

Global Gene Expression Profiling and Alternative Splicing Events during the Chondrogenic Differentiation of Human Cartilage Endplate-Derived Stem Cells.

Low back pain (LBP) is a very prevalent disease and degenerative disc diseases (DDDs) usually account for the LBP. However, the pathogenesis of DDDs is complicated and difficult to elucidate. Alternative splicing is a sophisticated regulatory process which greatly increases cellular complexity and phenotypic diversity of eukaryotic organisms. In addition, the cartilage endplate-derived stem cells have been discovered and identified by our research group. In this paper, we continue to investigate gene expression profiling and alternative splicing events during chondrogenic differentiation of cartilage endplate-derived stem cells. We adopted Affymetrix Human Transcriptome Array 2.0 (HTA 2.0) to compare the transcriptional and splicing changes between the control and differentiated samples. RT-PCR and quantitative PCR are used to validate the microarray results. The GO and KEGG pathway analysis was also performed. After bioinformatics analysis of the data, we detected 1953 differentially expressed genes. In terms of alternative splicing, the Splicing Index algorithm was used to select alternatively spliced genes. We detected 4411 alternatively spliced genes. GO and KEGG pathway analysis also revealed several functionally involved biological processes and signaling pathways. To our knowledge, this is the first study to investigate the alternative splicing mechanisms in chondrogenic differentiation of stem cells on a genome-wide scale.

Characterization of degenerative human facet joints and facet joint capsular tissues.

OBJECTIVE: Lumbar facet joint degeneration (FJD) may be an important cause of low back pain (LBP) and sciatica. The goal of this study was to characterize cellular alterations of inflammatory factor expression and neovascularization in human degenerative facet joint capsular (FJC) tissue. These alterations in FJC tissues in pain stimulation were also assessed. DESIGN: FJs were obtained from consented patients undergoing spinal reconstruction surgery and cadaveric donors with no history of back pain. Histological analyses of the FJs were performed. Cytokine antibody array and quantitative real-time polymerase chain reaction (qPCR) were used to determine the production of inflammatory cytokines, and western blotting analyses (WB) were used to assay for cartilage-degrading enzymes and pain mediators. Ex vivo rat dorsal root ganglion (DRG) co-culture with human FJC tissues was also performed. RESULTS: Increased neovascularization, inflammatory cell infiltration, and pain-related axonal-promoting factors were observed in degenerative FJCs surgically obtained from symptomatic subjects. Increased VEGF, (NGF/TrkA), and sensory neuronal distribution were also detected in degenerative FJC tissues from subjects with LBP. qPCR and WB results demonstrated highly upregulated inflammatory cytokines, pain mediators, and cartilage-degrading enzymes in degenerative FJCs. Results from ex vivo co-culture of the DRG and FJC tissue demonstrated that degenerative FJCs increased the expression of inflammatory pain molecules in the sensory neurons. CONCLUSION: Degenerative FJCs possess greatly increased inflammatory and angiogenic features, suggesting that these factors play an important role in the progression of FJD and serve as a link between joint degeneration and neurological stimulation of afferent pain fibers.

Association between estrogen receptor gene polymorphism and back pain intensity in female patients with degenerative lumbar spondylolisthesis.

STUDY DESIGN: Prospective study. OBJECTIVE: To examine the possible association of estrogen receptor α (ERα) polymorphisms and pain intensity in symptomatic female degenerative spondylolisthesis (DS) patients. SUMMARY OF BACKGROUND DATA: DS has been associated with a significant sex effect. Thus, several studies about the association between the ER gene and osteoarthritis have been reported. However, whether estrogen is associated with pain sensitivity is inconsistent in the existing literatures from both human and animal studies. METHODS: The PvuII and XbaI polymorphisms, bone mineral density at the lumbar spine (LSBMD) and at the femoral neck (FNBMD), pain intensity at the leg and lower back, and radiologic and anthropometric findings were analyzed in 192 patients with DS. RESULTS: There was a significant association between XbaI polymorphism and the visual analog scale score of back pain. The back pain visual analog scale in patients with a GG genotype was significantly higher than in patients with the AG (P<0.05) or the AA (P<0.05) genotypes. In addition, the presence of the CG haplotype was found to be associated with back pain intensity in the haplotype analysis of the PvuII and the XbaI polymorphisms of ERα. CONCLUSIONS: These results suggest that the ERα gene polymorphism using XbaI restriction enzyme influences the perception of back pain in patients with DS.

Type IX collagen neo-deposition in degenerative discs of surgical patients whether genotyped plus or minus for COL9 risk alleles.

STUDY DESIGN: Immunohistochemical analysis of type IX collagen in disc tissue from spinal fusion patients. OBJECTIVE: To determine if collagen IX can be detected in adult disc tissue removed at spinal fusion surgery from patients either with or without degeneration-associated tryptophan single nucleotide polymorphisms (SNPs) and whether the distribution is associated either with severity of degeneration or incidence of a collagen IX SNP genotype. SUMMARY OF BACKGROUND DATA: Genetic factors are strongly associated with risk of development and/or progression of disc degeneration. Two SNPs that introduce tryptophan polymorphisms in COL9A2 and COL9A3 are independently linked to an increased risk of lumbar disc disease. Although tryptophan variants are associated with accelerated degeneration, it is not known if collagen IX can be detected in adult disc tissue. METHODS: We selected age-matched disc samples from five clinical groups: fracture with Trp(-) (six cases), herniation (six cases), degeneration (five cases), spondylolisthesis with Trp(-) (eight cases), and spondylolisthesis/herniation/fracture with Trp(+) (six cases of Trp3 allele and one case of Trp2 allele). Using hematoxylin and eosin staining and immunohistochemical staining (collagens IX and IIA), 78 sections from 32 patients were analyzed. Selected disc tissues were assayed biochemically for collagen IX. RESULTS: Focal deposition of collagen IX was observed in regions of adult human disc tissue from spines showing degenerative changes in patients whether or not they were positive for a tryptophan SNP. However, in nondegenerative control disc tissue from fracture cases, little or no collagen IX was detected. The latter finding was confirmed by direct biochemical analyses for collagen IX in pooled samples of normal adult human annulus fibrosus or nucleus pulposus. CONCLUSION: During growth and maturation of the disc, collagen IX is presumably removed completely during matrix remodeling so that the protein is absent from normal adult annulus and nucleus but can reappear at sites of degeneration presumably as part of a repair response to mechanical injury.

Spondylolisthesis in twins: multifactorial etiology: a case report and review of the literature.

STUDY DESIGN: Report of a high dysplastic developmental spondylolisthesis in two identical twins of two unrelated families. OBJECTIVE: To investigate the multifactorial etiology of developmental spondylolisthesis. SUMMARY OF BACKGROUND DATA: Multiple studies have suggested an association between a high pelvic incidence and the presence of isthmic spondylolisthesis. Other studies suggest a genetic background for spondylolysis and a pattern of inheritance of susceptibility to spondylolysis and spondylolisthesis. Heterozygous cartilage-derived morphogenetic protein-1 (CDMP-1) mutation has previously been associated with spondylolysis and severe spondylolisthesis. METHODS: Two identical female twins presented with a developmental spondylolisthesis. Pelvic parameters, lumbar lordosis and grade of spondylolisthesis were calculated on a lateral standing spine radiograph. MRI is performed to confirm a high dysplastic developmental spondylolisthesis. Blood sample of these four individuals were analyzed for the presence of a CDMP-1 mutation, a cartilage-specific member of the TGF-b superfamily of secreted signaling molecules that plays a key role in chondrogenesis, growth, and patterning of the developing vertebrate skeleton. RESULTS: PI, SS, PT, LL, and SI are significantly greater in all of these patients in comparison with the general population. Spinal MRI confirms a high dysplastic developmental spondylolisthesis in both twins. Mutation analysis of the two coding exons of CDMP-1 did not reveal any mutation in all four individuals. CONCLUSION: To our knowledge, this is the first report of a high dysplastic developmental spondylolisthesis in identical twins. The presence of a high dysplastic developmental spondylolisthesis in two identical twins shows the convergence in etiology of different factors such as genetics, maturation, critical age, female sex, high pelvic incidence. Although we cannot confirm that CDMP-1 mutation plays a key role in the etiology of spondylolysis/spondylolisthesis, neither can we rule out that CDMP-1 problems may be an etiology for at least a subpopulation of patients. However, the presence of a developmental spondylolisthesis in two sets of identical twins still suggests a genetic susceptibility to spondylolysis and spondylolisthesis.

Matrix metalloproteinase-3 on ligamentum flavum in degenerative lumbar spondylolisthesis.

STUDY DESIGN: Human ligamentum flavum (LF) was examined for the activity level of matrix metalloproteinase-3 (MMP-3) in degenerative spondylolithesis (DS) patients using immunohistochemistry, Western blot, reverse transcriptase-polymerase chain reaction (RT-PCR), and quantitative real-time PCR. OBJECTIVE: To investigate the hypothesis that the activity of MMP-3 is elevated in LF of DS patients, which might contribute to DS pathogenesis. SUMMARY OF BACKGROUND DATA: MMP-3 is a proteinase produced by connective tissue cells and is responsible for the degradation and modification of extracellular matrix molecules. MMP-3 activity has been established in articular cartilage, synovial membrane, and intervertebral discs, but not in the LF. METHODS: The experimental group consisted of 18 patients with DS and the control group consisted of 18 patients with spinal stenosis (SS) without any instabilities. MMP-3 expression was measured with in situ using immunohistochemistry and both for mRNA and protein levels. RESULTS: The MMP-3 positive cell ratio in the LF observed in DS patients was substantially higher than in SS patients (P = 0.030). In Western blot, the average optical density (OD) of MMP-3 was higher in LF of DS than of SS (P = 0.028). There was greater MMP-3 expression in DS patients as quantified by RT-PCR (P = 0.004). CONCLUSION: Our study shows that MMP-3 expression in the LF of DS patients was significantly higher than in SS patients. Increased MMP-3 expression may be associated with the degenerative changes of LF in DS patients comprising one of the mechanisms of pathogenesis in DS.

Expression of estrogen receptor of the facet joints in degenerative spondylolisthesis.

STUDY DESIGN: Immunohistochemical study was done by harvesting articular cartilage of the facet joints during the decompressive surgery for spinal stenosis. OBJECTIVES: To observe the expression of estrogen receptor on the articular cartilage of the facet joints in degenerative spondylolisthesis (DS) SUMMARY OF BACKGROUND DATA: Few attempts have been made to evaluate the effect of sex-hormone, although DS is more common in females than in males. METHODS: After harvesting the articular cartilage of the facet joints in 17 DS and in 15 spinal stenosis (SS) patients, the expression of estrogen receptor and the severity of facet arthritis were observed by H-E and immunohistochemical staining, respectively. Measurements of both staining were made by using a semiquantitative analysis. RESULTS: The significantly increased expression of estrogen receptor correlated with the severity of facet arthritis (r = 0.78, P < 0.05). There was a significantly increased expression of estrogen receptor of the facet joint in DS compared with SS (P < 0.01). The histologic-histochemical grading of cartilage lesion in DS was 12.4 (SEM, 0.6), which was significantly higher than in SS (P < 0.05). CONCLUSIONS: These findings suggest that the higher expression of estrogen receptor might aggravate degenerative change of the facet articular cartilage and might also be considered one of the causative factors for DS in postmenopausal women.

The association of lumbar spondylolisthesis with collagen IX tryptophan alleles.

Two collagen type IX gene polymorphisms that introduce a tryptophan residue into the protein's triple-helical domain have been linked to an increased risk of lumbar disc disease. To determine whether a particular subset of symptomatic lumbar disease is specifically associated with these polymorphisms, we performed a prospective case-control study of 107 patients who underwent surgery of the lumbar spine. Patients were assigned to one of five clinical categories (fracture, disc degeneration, disc herniation, spinal stenosis without spondylolisthesis and spinal stenosis with spondylolisthesis) based on history, imaging results, and findings during surgery. Of the 11 tryptophan-positive patients, eight had spinal stenosis with spondylolisthesis and three had disc herniation. The presence of the tryptophan allele was significantly associated with African-American or Asian designation for race (odds ratio 4.61, 95% CI 0.63 to 25.35) and with the diagnosis of spinal stenosis with spondylolisthesis (odds ratio 6.81, 95% CI 1.47 to 41.95). Our findings indicate that tryptophan polymorphisms predispose carriers to the development of symptomatic spinal stenosis associated with spondylolisthesis which requires surgery.

Spondyloptosis and multiple-level spondylolysis.

An unusual case of a combination of multiple bilateral spondylolyses (L2, 3 and 4), spondylolisthesis at L3/4, spondyloptosis at L4/5 and sacralization of L5 in a teenage female is described. The patient had severely increasing lower back pain radiating to the left lower limb. Radiography identified the abnormalities and myelography revealed complete obstruction and compression of the thecal sac at the L4/5 level. The case was treated surgically by posterior decompression, corpectomy and fusion in a three-stage operation. The follow-up was extended to 2 years with no complications. No similar case has previously been reported.

L4-5 degenerative spondylolisthesis in monozygous twins.

OBJECTIVE: The author reports symptomatic spinal canal stenosis due to degenerative spondylolisthesis at L4-5 in identical twins. SUMMARY OF BACKGROUND DATA: The patient reported his brother had a similar condition. METHODS: Monozygosity was established by phenotyping. CONCLUSIONS: Spinal morphology and degenerative disease may be genetically determined.

Phenotypic variability of mannosidosis type II: report of two Greek siblings.

Two patients, a 13-year-old boy and his 24-year-old sister, were diagnosed as mannosidosis type II cases, on the basis of both presenting extremely reduced plasma and white blood cell acid-alpha-mannosidase are reported. With the exception of mental retardation and neurosensory deafness the two siblings manifested a wide phenotypic variability. The boy had several facial features indicating a lysosomal storage disorder, as well as spondylolisthesis. His sister, apart from heavy eyebrows and lower jaw prognathism appeared normal.

[Spondylolisthesis in athletes]. / Le spondylolisthésis du sportif.

Isthmic spondylolysis is a fatigue fracture which mostly affects athletes. It is acquired between the ages of 5 and 15 years and is directly related to the type of sport practised. Ethnic and racial factors are also involved. It is of interest to identify the first symptoms of isthmic damage at the initial stage. The practice of sports must be interrupted to facilitate isthmic repair. Sporting activities can be resumed, but the initial therapeutic procedure (hyperlordosis) must be modified. Once spondylolysis has been constituted, few sports are contra-indicated in adolescents. Major displacements towards spondyloptosis are rare, but surveillance is necessary. Isthmic spondylolysis is usually well tolerated by adults.

Monozygotic twins concordant for Rubinstein-Taybi syndrome.

Monozygotic twin sisters from Qatar, concordant for the Rubinstein-Taybi syndrome, are described. Skeletal anomalies not previously seen in this syndrome are described. The mode of inheritance is reviewed.

[Bone changes in Gaucher disease]. / Knochenveränderungen bei Morbus Gaucher.

Among Ashkenazi-Jews, Gaucher' disease, an autosomal-recessive hereditary genetic defect of sphingolipid metabolism, occurs more frequently than in the general population. Because of lack of the specific b-glucosidase, glucocerebrosidase, there is increased deposition of glucocerebrosides in the reticulo-endothelial system, mostly in the spleen, liver, and bone-marrow. In the chronic adult form (type 1), in addition to the hematologic complications (which are mostly associated with a splenomegaly), a less known involvement of the skeletal system occurs, which can lead to significant rheumatologic and orthopedic problems. These are: nonspecific skeletal and joint pain, purulent osteomyelitis, pseudo-osteomyelitis, aseptic necrosis of the femoral head, pathologic fractures of the long bones, acutely occurring kyphosis secondary to pathologic vertebral fractures with or without spinal compression, bony deformities, growth disturbances, arthritis, and bursitis. One sees a wide variety of bone lesions which can be solitary or multiple. Various pathogenic mechanisms have been discussed: toxic reaction to the Gaucher cells, disturbance of the osteoblastic and osteoclastic function, compression of osseous blood vessels by pathological cells, pressure-induced atrophy of the surrounding osseous tissue, local hemorrhage, local thrombosis, invasion of the arterioles with subsequent occlusion, and bone infarcts. The therapy is purely symptomatic. For orthopedic problems there is a greater tendency towards conservative treatment. There is disagreement as to whether splenectomy, which is often performed for hematologic or mechanical reasons, accelerates involvement of the bone. The case of a patient with multiple fractures of the spine and a slight spinal compression is presented.

Aspartylglucosaminuria in a Puerto Rican family: additional features of a panethnic disorder.

We report on 3 Puerto Rican brothers with the clinical and laboratory findings of aspartylglucosaminuria (AGU). Their parents were first cousins. The affected sibs have the "cardinal" manifestations of AGU, including developmental disabilities, progressive "coarsening" of the face, and early onset of hepatosplenomegaly. Biochemical studies showed elevated levels of urinary aspartylglucosamine and very low activity of aspartylglucosaminidase(AGA) in cultured fibroblasts. With long term follow-up, previously undescribed manifestations were noted, including radiographic evidence of spondylolysis and spondylolisthesis in early childhood and development of macro-orchidism during puberty. This family shows that AGU is not limited to individuals of Finnish background, but that the gene is panethnic in distribution and that additional changes, not previously noted, may present with advancing age.

[Concordant occurrence of grade I spondylolisthesis in childhood in a pair of monozygotic twins]. / Konkordantes Auftreten einer Spondylolisthesis Grad I im Kindesalter bei einem eineiigen Zwillingspaar.

The case of two identical female twins is presented, who both showed concordantly spondylolisthesis L 5/S 1 grade I according to Meyerding at the age of ten. Further abnormalities in spinal development were detected in both girls. The course of 31 months is reported.

Familial spondylolisthesis of the axis vertebra.

This report describes a nine-year-old girl with a spondylolisthesis of the C2 vertebra allowing 14 mm of slip. Her father had very similar vertebral anomalies.

Spondylolysis in Eskimo skeletons.

Spondylolysis was found in 25 of 46 spines of Eskimos from Greenland, at L1 in one case, at L3 in five cases and at L4 and L5 in 14 cases each. Two spondylolytic vertebrae were seen in the same spine in nine cases. Among young individuals, spondylolysis was found in 2 out of 15 spines and in older in 23 out of 31. The prevalence of spondylolysis was higher, the fourth vertebra was more commonly affected, and spondylolysis occurred at an older age in Eskimos than in other ethnic groups.