Anterior High-Intensity Zone in Lumbar Discs: Prevalence and Association with Low Back Pain.

OBJECTIVES: To explore the features of high-intensity zone (HIZ) in anterior annulus fibrosus and assess the association of anterior HIZ with low back pain (LBP). DESIGN, SETTING, AND SUBJECTS: A retrospective study of 5,940 discs in 1,188 individuals was conducted. METHODS: Subjects' information and LBP symptoms confirmed by an orthopedic surgeon were acquired from the medical record. Magnetic resonance (MR) image reading and analysis were performed by two experienced blinded radiologists. RESULTS: Two hundred eighty individuals exhibited 355 anterior HIZs in 355 discs. The prevalence was 23.57%; 88.45% were located in the inferior part of the annulus fibrosus. It frequently occurred in the middle and upper segments of lumbar spine, especially at L3/4 (45.63%). Of the 355 anterior HIZs, only 79 (22.25%) were consecutive-slides HIZ. Round type (63.38%) was the most common shape of anterior HIZs. The highest prevalence was found in individuals aged 60-69 years. LBP was confirmed in 141 anterior-HIZ individuals. The incidence of LBP in anterior-HIZ individuals was significantly higher than in non-HIZ subjects (50.36% vs 35.24%, χ2 = 18.314, P < 0.001). CONCLUSIONS: Anterior HIZ is a lower-prevalence, age-related sign on lumbar MR images. The spatial distribution of anterior HIZ can be distinguished from posterior HIZ. The number of consecutive anterior HIZ slides might suggest fewer Dallas grade 4 anterior annular disruptions in this sample. Anterior HIZ was correlated with LBP.

Thoracic intervertebral disc calcification and herniation in adults: a report of two cases.

PURPOSE: Acute paraplegia due to thoracic intervertebral disc protrusion and calcification is rare. The purpose of this study was to report two cases with acute paraplegia due to a calcified thoracic disc prolapse, and discuss its clinical diagnosis and surgical treatment with literature reviews. METHODS: These two cases were verified by patient history, physical examination, laboratory examination, CT and MRI studies, and pathological findings. RESULTS: CT scan revealed disc calcification and protrusion at the T11-12 level in case 1 and at the T10-11 level in case 2, respectively. MRI images revealed severe spinal cord compression with a hyperintense central core and surrounding hypointense area in two cases, which were directly connected to the calcified intervertebral nucleus pulposus. Pathological examination revealed calcium deposition. Patients underwent discectomy followed by interbody fusion, and satisfactory therapeutic outcomes were obtained. CONCLUSIONS: We suggest that decompression surgery should be carried out as early as possible for patients with early spinal myelopathy or paraplegia caused by a calcified protruded disc.

[A novel rabbit disc degeneration model induced by fibronectin fragment].

OBJECTIVE: To establish a novel and useful rabbit model of lumbar disc degeneration using microinjection of fibronectin fragment (Fn-f). METHODS: Thirty-two New Zealand white rabbits underwent injection of N-terminal 30 kDa Fn-f (experimental group) or phosphate buffered saline (PBS) (control group) into the central region of L1-2, L2-3, L3-4, L4-5 discs using a 32-gauge microsyringe. Two rabbits (blank group) with no treatments were sacrificed to examine the proteoglycan synthesis of neucleus pulposus (NP) using (35)S-sulfate incorporation assay. At the 4-, 8-, 12-, and 16-week time points, the discs were examined histologically, radiographically, and with proteoglycan synthesis. RESULTS: Histology demonstrated a progressive loss of the cell numbers in NP and architecture destruction in NP and anulus fibrosus (AF) in Fn-f-injected discs over the 16-week study period. The NP regions in Fn-f-injected discs shrinked distinctly after the 4-week time point, and were not discernible with the inner AF by the 16-week time point. Protoglycan synthesis in Fn-f-injected discs decreased progressively (F = 263.241, P = 0.000). At each time point, the Fn-f-injected discs showed significantly decreased proteoglycan synthesis compared with controls (t = -27.010 - -2.833, P < 0.05). The DHI% of the Fn-f-injected discs at the 4-, 8-, 12-, and 16-week time points were 96.5% ± 1.7%, 85.6% ± 3.8%, 77.2% ± 3.5% and 65.5% ± 5.6%, respectively. Comparing with the DHI% of PBS-injected discs (97.4% ± 1.2%), the Fn-f-injected discs exihibited no significant differences in disc heights at the 4-week time point (P > 0.05), but significant decreases in disc heights at the 8-, 12-, and 16-week time points (t = -21.225 - -10.795, P < 0.01). Apparent anterior osteophytes formed at the 12-week time point and enlarged remarkablely by the 16-week time point in the experimental spines. CONCLUSIONS: Fn-f can induce a progressively degenerative process in rabbit discs which is ethical, cost-effective, reproducible, and consistent with the spontaneous degeneration in human. And it seem to be a novel and useful model for the study of disc degeneration at the molecular level.

[The changes of extracellular matrix in adult degenerative nucleus pulposus cells with stiring microcarrier culture system in vitro].

OBJECTIVE: To evaluate the biological effect on the synthesis of the extracellular matrix (ECM) in the cultivation of adult degenerative nucleus pulposus cells using the stiring microcarrier system in vitro. METHODS: Thirty-four specimens were collected after intervertebral fusion operations of the patients with intervertebral disc herniation diseases from September 2005 to May 2009. The specimens were then randomly allocated into 2 groups for in vitro cultivation: monolayer culture group and microcarrier culture group. On the exponential phase, SP-ABC immunohistochemical staining and Western blot quantitative analysis were conducted in the two groups to detect the collagen type I and II. Proteoglycan contents of two groups in different growth phases were detected with (35)S-sulfate incorporation assay. RESULT: The expressions of collagen type I and II in microcarrier culture group were significantly higher than those in monolayer culture group: SP-ABC immunohistochemical staining (collagen type I: 32.5 ± 4.4 vs. 15.2 ± 1.2, t = 2.871, P < 0.01; collagen type II: 43.6 ± 4.1 vs. 23.1 ± 2.2, t = 2.375, P < 0.05); Western blot quantitative analysis (collagen type I: 0.62 ± 0.08 vs. 0.50 ± 0.06, t = 3.327, P < 0.01; collagen type II: 1.46 ± 0.08 vs. 0.86 ± 0.04, t = 2.453, P < 0.05). Nucleus pulposus cells cultivated in stiring microcarrier system showed significantly increased proteoglycan synthesis than monolayer culture group does on both exponential phase and stationary phase (exponential phase: 34 821 ± 312 vs. 21 046 ± 673, t = 2.134, P < 0.05; stationary phase: 45 134 ± 175 vs. 32 193 ± 713, t = 2.801, P < 0.01). CONCLUSIONS: The expression of collagen type I, II and proteoglycan of adult degenerative nucleus pulposus cells are positive regulated by the stiring microcarrier system, which can be used in the mass amplification of the adult degenerative nucleus pulposus cells.

Correction of post-traumatic thoracolumbar kyphosis using pedicle subtraction osteotomy.

The aim of this study was to retrospectively analyze and evaluate the effect of treatment employing pedicle subtraction osteotomy for chronic, posttraumatic thoracolumbar kyphosis. This study included 19 patients, 11 males and 8 females, with chronic, posttraumatic thoracolumbar kyphosis. Pre-operative kyphosis ranged from 31° to 63°. The history of trauma ranged from 8 to 63 months. All patients were treated with pedicle subtraction osteotomy. A mean 40.2° improvement in sagittal alignment was achieved with a mean correction rate of 85.8 %. Perioperative complications were encountered in two patients, one with cerebrospinal fluid leakage followed by encephalitic infection and one with a wound infection. Both were treated conservatively with antibiotics and local wound care. There were no other severe complications. The average follow-up period was 15 months (range 6-41 months). At the last follow-up, clinical symptoms and neurological function were significantly improved. Of 14 patients presenting with intractable back pain, VAS scores improved from a preoperative mean of 6.7 (range 5.0-8.0) to an average 2.0 (range 0-3.0) at final follow-up. No significant loss of correction was observed (loss of 1.7°), and solid fusion was achieved in all 19 patients. A single-stage posterior pedicle subtraction osteotomy is a safe and effective procedure for correction of posttraumatic thoracolumbar kyphosis. Using this technique, it is possible to safely obtain no greater than 55° of correction at a single level.

High-intensity zone (HIZ) of lumbar intervertebral disc on T2-weighted magnetic resonance images: spatial distribution, and correlation of distribution with low back pain (LBP).

OBJECTIVES: To analyze the three-dimensional distribution of high-intensity zone (HIZ) in lumbar disks and to assess the correlation between low back pain (LBP) and spatial distribution of HIZs. METHODS: Clinical records and lumbar MRIs of 623 patients (337 males and 286 females, age 50.10 ± 15.38 years) were selected and reviewed. Baseline characteristics and 3D localization were performed and recorded by two radiologists in a blind fashion. RESULTS: Among the 623 patients, 200 exhibited an HIZ in at least one disk. HIZs were more frequently seen in the inferior part of annulus fibrosus (superior-middle-inferior ratio 39:59:140, P < 0.001). One hundred and eighty-one HIZs (76.1%) occurred at L4/5 and/or L5/S1. The prevalence of multi-segmental HIZ was 16.5%. Among the 33 patients with multi-segmental HIZs, 24 exhibited HIZs in adjacent disks. The LBP rate of HIZ patients was significantly higher than that of patients who exhibited no HIZ (57.5 vs. 47.8%, P < 0.05). There was no evidence for a correlation between LBP and spatial distribution of HIZ in disk (P > 0.05). The incidence of LBP was slightly higher when the HIZ disk level was lower or when there were HIZs exhibited in more disks; however, the difference was statistically insignificant (P > 0.05). CONCLUSIONS: High-intensity zones occurred frequently at lower segments, inferior part of annulus fibrosus, and single disk.

Imaging Analysis of the High-Intensity Zone on Lumbar Spine Magnetic Resonance Images: Classification, Features and Correlation with Low Back Pain.

AIM: Early studies suggested that the high-intensity zone (HIZ) on lumbar MRI was a diagnostic sign of painful internal disc disruption (IDD). However, recent studies have questioned its diagnostic value. This study is conducted to explore imaging features of HIZ and to investigate the correlation between these characteristics and low back pain (LBP), further studying the predictive value of HIZ. METHODS: A retrospective study of 1188 cases was performed. MR images were read and analyzed by two experienced, blinded radiologists. RESULTS: A total of 575 (48.4%) individuals exhibited HIZ. The prevalence of posterior HIZ (32.3%) was significantly higher than that of anterior HIZ (23.6%; P < 0.01). Round type was the most common shape (61.0%) on sagittal view. Only 37 HIZs (4.6%) were identified on axial views. A total of 263 HIZ discs (32.5%) were found to have additional diagnostic signs of IDD, which is difficult to distinguish from the annulus fibrosus. In subjects with consecutive slides showing HIZ, the incidence of LBP was significantly higher than in single-slide HIZ individuals (58.0% vs 48.6%, P < 0.05). CONCLUSION: MRI-visualized HIZ is a highly valuable method of screening for lumbar IDD. It is demonstrated that consecutive-slide HIZ was a more reliable indicator for discogenic LBP than single-slide HIZ.

[Diagnosis and treatment of disc herniation at cervical vertebra 7-thoracic vertebra 1 (C(7)-T(1))].

OBJECTIVE: To study the outcome of laminoforaminotomy with posterolateral discectomy for patients with lateral disc herniation at C(7)-T(1). METHODS: From August 2000 to August 2008, 12 patients with lateral disc herniation at C(7)-T(1) underwent posterolateral discectomy were analyzed retrospectively. Neurologic function were evaluated with the Motor Scoring System. Preoperative motor were compared with postoperative one. The unique clinical manifestation, imageology features and intraoperative findings were analyzed. RESULTS: All these twelve patients were lateral type. All the patients showed hand intrinsic muscles atrophy and hand weakness. Nine patients had no paraesthesia. The average follow-up period was 26 months. Postoperative scores were significantly higher than preoperative ones. CONCLUSIONS: Disc herniation at C(7)-T(1) is predominantly lateral type and present C(8) nerve motor deficit (hand intrinsic muscles atrophy and hand weakness) and only minority has paraesthesia in C(8) nerve dermatome. Posterolateral cervical discectomy technique is safe and effective for patients with lateral disc herniation at C(7)-T(1).

[Clinical investigation of high-intensity zone in anterior annulus fibrosus of lumbar disc: compared with high-intensity zone in posterior annulus fibrosus].

OBJECTIVE: To explore the prevalence, distribution and clinical significance of high-intensity zone (HIZ) in anterior annulus fibrosus (AF) in comparison with HIZ in posterior AF of lumbar disc. METHODS: According to the diagnosis and location of HIZ, 610 lumbar magnetic resonance images with entire clinical materials were divided into control group (without HIZ), anterior AF group (HIZ), posterior AF group (HIZ) and anterior & posterior (AP) AF group (HIZ). The incidence of HIZ was summarized. The clinical data, such as male/female ratio, age, and body weight, prevalence of low back pain (LBP) and distribution of HIZ, were compared and analyzed between the groups. RESULTS: Three hundred and fifteen cases shown no HIZ (51.6%), 95 cases presented HIZ in anterior AF (15.6%, 119 discs), 159 cases presented HIZ in posterior AF (26.1%, 189 discs) and 41 cases presented HIZs in both anterior and posterior AF (6.7%, 96 discs). There was significant difference between the prevalence of HIZ in anterior AF and that in posterior AF (P < 0.01). The male/female ratio and body weight of each groups showed no difference (P > 0.05), and the age was proved to be statistically different between four groups (P < 0.01, control group < posterior AF group < AP AF group < anterior AF group). HIZs in anterior AF often occurred at L(1,2)-L(4,5), whereas, they usually developed at L(3,4)-L(5)S(1) in posterior AF. The incidence of LBP in control group, anterior AF group, posterior AF group, AP AF group were 40.0%, 52.6%, 55.4% and 65.8%, respectively. The LBP prevalence of control group was lower than that of other three groups (P < 0.05), and the prevalence of last three groups showed no difference (P > 0.05). CONCLUSIONS: Compared with HIZ in posterior AF, the HIZ in anterior AF of lumbar disc has a lower prevalence, often develops in elder patients and in upper motion segments. It also indicates an obvious relation to LBP as the former.

[Outcomes of posterior C(1-2) fusion for atlantoaxial instability or dislocation using pedicle screws of atlas and axis].

OBJECTIVE: To retrospectively analyze and evaluate the results of treatment for atlantoaxial instability or dislocation employing pedicle screws of atlas and axis. METHODS: Thirty-one patients (23 male and 8 female) with atlantoaxial instability or dislocation were stabilized using pedicle screws of atlas and axis between May 2005 to January 2008. The patients ranged in age from 17 to 67 years (mean 43.5 years). Patients consisted of chronic odontoid fracture in 17, Os odontoideum in 8, fresh odontoid fracture in 4, transverse ligament rupture in 1, rheumatoid arthritis in 1. Clinical features included neck pain in 31; restricted neck movement in 28, varying degrees of spastic quadriparesis in 19. All patients underwent posterior C(1) to C(2) pedicle screw fixation. Operative time, intraoperative blood loss, complications were recorded, neurological and radiographic studies were carried. RESULTS: Mean follow-up time was 13 months. Operative time averaged 2.5 h. Mean intraoperative blood loss was 300 ml. A patient had postoperative wound infection and was treated conservatively with antibiotics and local wound care. A patient developed pulmonary artery embolism and got well with anticoagulation. Satisfactory stability was achieved in all cases with no vascular and C(2) neuralgia. Average JOA score in 19 cases increased at final follow-up (P < 0.01). Solid fusion was achieved in 29 cases, fusion rate was 93.6%. CONCLUSIONS: Stabilization of atlantoaxial complex via pedicle screws of atlas and axis has advantages of intraoperative restoration, easier placement of screw, solid fixation. It is a safe and effective treatment modality for posterior C(1-2) fusion.

The research of transgenic human nucleus pulposus cell transplantation in the treatment of lumbar disc degeneration.

The present study determines whether the in vivo injection of TGFß1 and CTGF mediated by AAV2 to transfect nucleus pulposus cells in degenerative lumbar discs can reverse the biological effects of rhesus lumbar disc degeneration. A total of 42 lumbar discs obtained from six rhesus monkeys were classified into three groups: experimental group, control group, and blank group. Degenerative lumbar discs were respectively injected with double gene-transfected human nucleus pulposus cells using minimally invasive techniques. Immumohistochemical staining, RT-PCR, and western blot were performed to observe the biological effects of double gene-transfected human nucleus pulposus cells in degenerative lumbar discs on rhesus lumbar disc degeneration. At 4, 8, and 12 weeks after the transplantation of nucleus pulposus cells, the expression levels of TGF-ß1, CTGF, proteoglycan mRNA, and type-II collagen were detected by RT-PCR. The values of immumohistochemical staining and RT-PCR in the experimental group increased at 8 weeks, decreased with time at 12 weeks, and remained greater than the values in the control group, and the differences were statistically significant (P < .05). The western blot revealed that the values in the experimental group decreased with time, but remained greater than those in the PBS control group and blank control group, and the differences were statistically significant (P < .05). The double gene-transfection of human nucleus pulposus cells in degenerative lumbar discs mediated by rAAV2 can be continuously expressed in vivo after transplantation in lumbar discs of rhesus monkeys, and promotes the synthesis of proteoglycan and type II collagen, achieving the treatment purpose.

[Significance of the high-intensity zone located in the posterior annulus fibrosus for diagnosing discogenic low back pain].

OBJECTIVE: To investigate the prevalence of high-intensity zone (HIZ) and the correlation between HIZ and low back pain (LBP). METHODS: The data of 1000 unselected cases, 566 males and 434 females, aged 49.49 (12 - 86), who underwent lumbar MRI were analyzed to examine the prevalence of HIZ. 200 of the 1000 cases with HIZ which had complete clinical history records, 112 males and 88 females, aged 52.29 (18 - 86), were divided into LBP (n = 115) and non-LBP series (n = 85). CT discography (CTD) was performed in 16 discs in 11 cases with HIZ and the correlation between the modified Dallas' discogram scale and pain reproduction in provocation discography was explored. RESULTS: HIZ located in posterior annulus fibrosus (AF) was shown in 378 discs in 317 cases, most located in L4/5 and L5/S1 (74.8%) and the location in left side (153 HIZs) was more common than that in right side (94 HIZs). HIZs were shown in superior region (15.4%), middle region (28.8%), as well as inferior region (55.8%) in sagittal plane. 115 of the 200 cases with HIZ who had complete clinical history records (57.5%) were symptomatic and 85 cases (42.5%) were asymptomatic. According to the modified Dallas' discogram scale, LBP could be provoked in 8 of the 9 patients with grade IV and could not be provoked in 6 of the 7 patients with the grade III. CONCLUSION: Whether HIZ is accompanied with LBP is related to the degree of disc degeneration. When CTD showed the degree of modified Dallas' grade IV and over most of the patients will show LBP. HIZ only indicates the possibility of discogenic LBP and can not replace provocation discography.

[A model of lumbar disc degeneration on the early stage in rhesus monkey with minimally invasive technique].

OBJECTIVE: To establish a novel model of lumbar disc degeneration on the early stage in the rhesus monkey using percutaneous needle puncture guided by CT. METHODS: (1) Thirteen rhesus monkeys aged from 4 to 7 years, female 7 and male 6 were selected for establishing a model of the early stage of lumbar disc degeneration. (2)13 monkeys, 91 discs were divided into 3 groups: 64 discs from L1/2 to L5/6 were percutaneous punctured with a needle 20G as experimental group and 1 disc with a needle 15G as puncture control group and 26 discs were not be punctured from L6,7 to L7-S1 as control group. (3) Lumbar disc localization for needle puncture was guided by CT. All discs were examined by MRI, the HE, Masson's trichrome, Safranine-O and immunohistochemical staining of type II collagen before disc puncture and after puncture at 4, 8 and 12 weeks. RESULTS: MRI: (1) Experimental group: Pfirmann's Grade I was shown at postoperation 4, 8 and 12 weeks; (2) Puncture control group: Grade III was shown at postoperation 4 weeks and Grade IV at 8 weeks; (3) CONTROL GROUP: Grade I was shown at postoperation 4, 8 and 12 weeks. Histological examination: (1) In experimental group, there was no any change at postoperation 4 weeks, and the cell population of the nucleus was decreased at 8 weeks and more decreased at 12 weeks in HE. (2) There was no any change at postoperation 4 weeks, the clefts among the lamellae of the annulus fibrosus (AF) were shown at 8 weeks and more wider of the clefts of AF at 12 weeks in Masson's trichrome. (3) No any change was shown at postoperation 4 weeks, proteoglycan were progressively decreased at 8 and 12 weeks in Safranine-O. (4) No statistically significant difference in positive rate was observed at 4 and 8 weeks compared with control group in immunohistochemical staining of type II collagen. There was statistical difference at 12 weeks compared with control group (P<0.05). In puncture control group postoperation 8 weeks, the morphology of cell of nucleus pulposus was not clear in HE. The wider clefts of lamellae of the AF were shown in Masson's trichrome. The proteoglycan was obviously decreased in Safranine-O. Immunohistochemical staining collagen II synthesized was decreased. In normal control group, no any change was shown at 4, 8 and 12 weeks. CONCLUSIONS: The degeneration of lumbar intervertebral disc on the early stage could be induced by the percutaneous needle puncture (20G) to the annulus fibrosus. The assessment of disc degeneration on early stage is not shown on MRI and only confirmed by histological examination.

Factors associated with lumbar disc high-intensity zone (HIZ) on T2-weighted magnetic resonance image: a retrospective study of 3185 discs in 637 patients.

BACKGROUND: It is well known that internal disc disruption (IDD) is accelerated by factors such as aging and injury. High- intensity zone (HIZ) on lumbar MRI is usually considered a marker of painful IDD. However, many painful IDD show no HIZ. This suggests that the risk factors of HIZ may be different to these of IDD. The purpose was to clarify the correlation between the HIZ on lumbar MR and the factors, including gender, age, body weight, and low back pain (LBP). METHODS: Characteristics were obtained from the medical record. The MR images, biplanar post-discography radiographs, and post-discography CT images were reviewed and rated by two experienced radiologists in a blinded fashion. RESULTS: Annular HIZ correlated significantly with age (OR = 1.011), body weight (OR = 1.022), and LBP symptom (OR = 1.527). The lowest two HIZ prevalence rates were in the second and the third decades (11.54% and 7.84%). The highest prevalence was in the sixth decade (38.03%). The body weight was positively associated with the HIZ prevalence. There was a significant difference in HIZ prevalence between symptomatic and asymptomatic patients (36.16% vs. 26.96%, P < 0.05). All the HIZ discs exhibited grade 3 or grade 4 disruptions, but only 9 discs (9/16, 8 exhibited grade 4 annular tears) were detected with exact pain reproduction. CONCLUSIONS: It is demonstrated that the presence of HIZ on lumbar MR image was associated with aging, high body weight, and low back pain symptom. HIZ sign indicated a part of the natural history of disc degeneration but was not an actual source of low back pain.

Effects of AAV2-mediated co-transfection of CTGF and TIMP1 genes on degenerative lumbar intervertebral discs in rhesus monkeys in vivo.

OBJECTIVE: This study aims to investigate the effects of co-transfection of the genes for connective tissue growth factor (CTGF) and tissue inhibitor of metalloproteinase-1 (TIMP1) mediated by adeno-associated virus 2 (AAV2) on degenerative lumbar intervertebral discs in a primate model. METHODS: Twelve 4-7 year-old rhesus monkeys weighing 4.5-7.0 kg were utilized. CTGF and TIMP1 genes carried by AAV2 were injected into the degenerative lumbar intervertebral discs. Cytokine expression and biological effects were determined using quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and 35S-sulfate incorporation assays. A rhesus monkey model of intervertebral disc degeneration was successfully established. RESULTS: At post-transfection, CTGF mRNA expression was higher in the transfection group than in the control group (P < 0.05). Furthermore, TIMP1 mRNA expression in the transfection group was several times the levels observed in the control group (P < 0.05). Moreover, type-II collagen mRNA expression was higher in the transfection group than in the control group (P < 0.05). In addition, higher aggrecan mRNA expression and synthesis were observed in the transfection group, compared to that in the control group (P < 0.05). CONCLUSION: The stable expression of CTGF and TIMP1 genes in vivo promoted the synthesis of aggrecan and type II collagen in the nucleus pulposus in the rhesus monkey model of intervertebral disc degeneration, which has a potential for intervertebral disc regeneration.

The upper thoracic spinal cord compression as the initial manifestation of Wegener's granulomatosis: a case report.

There are only a few reported cases about spinal cord involvement with Wegener's granulomatosis (WG) in the literature. In these cases, the spinal cord is usually indented or compressed by dural and meningeal masses which are characterized by necrotizing granuloma formation and vasculitis. And, it usually cannot be correctly diagnosed. A 53-year-old woman suffered from Wegener's granulomatosis, in whom the upper thoracic spinal cord compression is the initial manifestation. The surgical biopsy and thoracic laminectomy were performed and the histologic examination was done. This patient was finally diagnosed as WG when the pathologic examination revealed as Wegener's granulomatosis and the serum antineutrophil cytoplasmic antibodies (ANCA) were reported positive; titers of antimyeloperoxidase (MPO) antibodies were markedly elevated. After treatment with cyclophosphamide and corticosteroids this patient partially recovered from neurological involvement. In a case such as this, careful monitoring of clinical parameters is essential for assessing disease activity with repeated MRI if neurologic status changes. Serial measurement of ANCA titers may also be helpful to establish the diagnosis. Cyclophosphamide and corticosteroids are the agents of choice for induction of remission of WG.

Constructing adeno-associated virus-TGFbeta3 and comparing its biological effect on proteoglycan synthesis in dedifferentiated nucleus pulpous cells with adenovirus-TGFbeta1.

OBJECTIVE: To construct adeno-associated virus (AAV) expression system for transforming growth factor beta3 (TGFbeta3 ) and detect its biological effect on proteoglycan synthesis of the earlier and later dedifferentiated rabbit lumbar disc nucleus pulpous (NP) cells, which was compared with that of adenovirus (AV) expression system for TGFbeta1. METHODS: TGFbeta3 gene was obtained using PCR. Its upstream contained restriction enzyme site Kpn I, and its downstream contained restriction enzyme site Sal I. Using the restriction enzyme sites of PCR product of TGFbeta3 and the corresponding multiple cloning site (MCS) in plasmid AAV, TGFbeta3 was subcloned into AAV. The recombinant plasmid AAV-TGFbeta3 was transfected into H293 cells with Lipofectamine 2000, and the expression of TGFbeta3 gene was detected using immunofluorescent analysis. After AAV-TGFbeta3 virus particle with infectious activity was packaged, TGFbeta3 expression in NP cells was detected by immunoblotting, and its biological effect on proteoglycan synthesis was detected by antonopulos method and compared with that of AV-TGFbeta1 in the earlier and later dedifferentiated NP cells. RESULTS: For the earlier dedifferentiated NP cells, AAV-TGFbeta3 slowly and stably enhanced proteoglycan synthesis, but AV-TGFbeta1 rapidly and transiently enhanced its synthesis. For the later dedifferentiated NP cells, AAV-TGFbeta3 stably enhanced proteoglycan synthesis, but AV-TGFbeta1 inhibited its synthesis. CONCLUSION: AAV expression system can mediate TGFbeta3 gene to be expressed stably, and AAV-TGFbeta3 can enhance proteoglycan synthesis of the earlier and later dedifferentiated NP cells.

Lentivirus-mediated TGF-ß3, CTGF and TIMP1 gene transduction as a gene therapy for intervertebral disc degeneration in an in vivo rabbit model.

The present study examined the effects of transforming growth factor (TGF)-ß3, connective tissue growth factor (CTGF) and tissue inhibitor of metalloproteinase 1 (TIMP1) gene transduction, using a lentiviral vector, on rabbit intervertebral disc degeneration in vivo, with the intention of investigating their potential use in gene therapy. A model of lumbar intervertebral disc degeneration was created by needle puncture into the annulus fibrosus of 15 New Zealand white rabbits. Empty lentivirus or recombinant lentiviral plasmid lenti-TGFß3-P2A-CTGF-T2A-TIMP1 was injected into degenerative lumbar intervertebral discs (representing the control and experimental groups, respectively), whilst untreated degenerative lumbar intervertebral discs served as the puncture group. After 16 and 20 weeks, magnetic resonance imaging (MRI) was conducted and the changes in intensity on micrographs of degenerative intervertebral discs were measured. The mRNA levels of aggrecan and type II collagen in nucleus pulposus tissue were determined by reverse transcription-polymerase chain reaction, and protein expression levels of type II collagen and aggrecan were determined by western blot analysis. MRI results indicated that intervertebral disc degeneration was ameliorated in the experimental group when compared with the control and the puncture group. Furthermore, the expression levels of type II collagen and aggrecan in the puncture and control groups were significantly lower than in the experimental group (P<0.05). In conclusion, lenti-TGFß3-P2A-CTGF-T2A-TIMP1 co-transduction can promote synthesis of aggrecan and type II collagen in degenerative intervertebral discs, thereby delaying intervertebral disc degeneration. These results indicate the potential of gene therapy in treatment of intervertebral disc degeneration.

Effect of rAAV2-hTGFß1 Gene Transfer on Matrix Synthesis in an In Vivo Rabbit Disk Degeneration Model.

STUDY DESIGN: In vivo gene transfer for disk regeneration. OBJECTIVE: To evaluate the efficiency and effect of human transforming growth factor ß1 (hTGFß1) gene transfer mediated by adeno-associated virus (AAV) in a rabbit disk degeneration model induced by fibronectin fragment (Fn-f). SUMMARY OF BACKGROUND DATA: Gene therapy for disk degeneration has been reported to be effective. Nevertheless, few investigations have targeted the degenerative nucleus pulposus (NP) cells in vivo. Fn-f-induced degeneration has been previously verified to be a useful model for the study of disk degeneration at the molecular level. AAV vector is well suited for gene transfer in the disk for its lower immunogenicity and higher safety. MATERIALS AND METHODS: The early dedifferentiated NP cells were transfected with rAAV2-mediated enhanced green fluorescent protein (EGFP) gene in vitro. Fluorescence expression was observed 48 hours later. The rabbit disk degeneration model was established with a microinjection of Fn-f. Ninety-six degenerative disks of 24 rabbits were injected with rAAV2-hTGFß1 (group A), rAAV2-EGFP (group B), or PBS (group C). Immunohistochemical staining for hTGFß1 and fluorescence observation were performed at the 1- and 12-week time points, respectively. 35S-sulfate incorporation assay and Western blot analysis were used to measure the synthesis of proteoglycan and collagen type II at 4-, 8-, and 12-week time points. RESULTS: The dedifferentiated NP cells exhibited intensive fluorescence expression in vitro, with a transfection rate of 90%. In vivo, disks in group A showed enhanced positive hTGFß1 immunostaining at the 1-week time point. At the 4-, 8-, and 12-week time points, disks in group A exhibited significantly increased proteoglycan and collagen type II synthesis compared with the other 2 groups (P<0.01). Abundant green fluorescence was observed in the disks in group B at the 12-week time point. CONCLUSIONS: Early degenerative NP cells are susceptible to AAV-mediated gene transfer in vitro and in vivo. The rapid and prolonged target protein expressions and increased matrix synthesis indicated that AAV-mediated therapeutic gene transfer can be a promising form of treatment for disk regeneration in vivo.

One stage laminoplasty and posterior herniotomy for the treatment of myelopathy caused by cervical stenosis with cervical disc herniation.

The aim of the study was to introduce a method of one stage laminoplasty and posterior herniotomy for myelopathy caused by cervical stenosis with cervical disc herniation and to evaluate the clinical efficacy of this surgery. From 1999 to 2008, 18 patients with myelopathy caused by cervical stenosis with cervical disc herniation who underwent this procedure were included. The average age was 63 years (range 48-74 years), and the average follow-up period was 46 months (range 3-108 months). Neurologic status was evaluated using the JOA scoring system. Neurological symptoms improvement was seen in all patients after surgery. The average JOA score was 14.22±1.86 by final follow-up, which was higher than preoperative values (P<0.01), and the average improvement in neurological function was 76.63%. Neurologic examination showed that excellent results had been obtained by 10 patients, good results by 8 patients, with no fair or poor results. 2 patients developed cerebrospinal fluid leakage after surgery and recovered during the follow-up period. One patient with cervical disc herniation developed postoperative C5 palsy on the axle side on the third day after surgery. She completely recovered by 1 month after surgery. No other patients experienced postoperative neurologic complications. Complete anterior and posterior decompression of the spinal cord was achieved after surgery. We concluded that one stage laminoplasty and posterior herniotomy is an effective, reliable, and safe procedure for the treatment of myelopathy caused by cervical stenosis with cervical disc herniation.