Clinically Feasible Microstructural MRI to Quantify Cervical Spinal Cord Tissue Injury Using DTI, MT, and T2*-Weighted Imaging: Assessment of Normative Data and Reliability.

BACKGROUND AND PURPOSE: DTI, magnetization transfer, T2*-weighted imaging, and cross-sectional area can quantify aspects of spinal cord microstructure. However, clinical adoption remains elusive due to complex acquisitions, cumbersome analysis, limited reliability, and wide ranges of normal values. We propose a simple multiparametric protocol with automated analysis and report normative data, analysis of confounding variables, and reliability. MATERIALS AND METHODS: Forty healthy subjects underwent T2WI, DTI, magnetization transfer, and T2*WI at 3T in <35 minutes using standard hardware and pulse sequences. Cross-sectional area, fractional anisotropy, magnetization transfer ratio, and T2*WI WM/GM signal intensity ratio were calculated. Relationships between MR imaging metrics and age, sex, height, weight, cervical cord length, and rostrocaudal level were analyzed. Test-retest coefficient of variation measured reliability in 24 DTI, 17 magnetization transfer, and 16 T2*WI datasets. DTI with and without cardiac triggering was compared in 10 subjects. RESULTS: T2*WI WM/GM showed lower intersubject coefficient of variation (3.5%) compared with magnetization transfer ratio (5.8%), fractional anisotropy (6.0%), and cross-sectional area (12.2%). Linear correction of cross-sectional area with cervical cord length, fractional anisotropy with age, and magnetization transfer ratio with age and height led to decreased coefficients of variation (4.8%, 5.4%, and 10.2%, respectively). Acceptable reliability was achieved for all metrics/levels (test-retest coefficient of variation < 5%), with T2*WI WM/GM comparing favorably with fractional anisotropy and magnetization transfer ratio. DTI with and without cardiac triggering showed no significant differences for fractional anisotropy and test-retest coefficient of variation. CONCLUSIONS: Reliable multiparametric assessment of spinal cord microstructure is possible by using clinically suitable methods. These results establish normalization procedures and pave the way for clinical studies, with the potential for improving diagnostics, objectively monitoring disease progression, and predicting outcomes in spinal pathologies.

[Effect of Acupotomy Lysis at Cervical Acupoints on Expression of Matrix Metalloproteinases and Tissue Inhibitor of Metalloproteinase 1 and Changes of Pulpiform Nucleus Ultrastructure in Rats with Degenerated Cervical Intervertebral Discs].

OBJECTIVE: To observe the effect of acupotomy lysis at acupoints around the neck on expression of matrix metalloproteinase 1 (MMP-1), MMP-2 and tissue inhibitor of metalloproteinase 1 (TIMP-1) genes and ultrastructure of pulpiform nucleus in cervical intervertebral disc degeneration (IVDD) rats, so as to explore its mechanism underlying easing IVDD. METHODS: SD rats were randomly allocated to control (n = 15), model (n = 14), Jiaji (EX-B 2, n = 13), cervico-acupoint (n = 14) and medication groups (n = 14). The cervical IVDD model was established by using static-dynamic imbalance method. For rats of the Jiaji (EX-B 2) and cervico-acupoint groups, EX-B 2-points of the cervical 2-7 segments, and peri-cervical acupoints: bilateral "Naokong" (GB 19) , "Naohu" (GV 17), "Dazhui" (GV 14), bilateral "Quyuan" (SI 13) and bilateral "Tianzong" (SI 11) were separately punctured with a needle-knife, once every 5 days for 3 times, and for rats of the medication group, Brufen Capsules (15 mg · kg(-1) · d(-1)) and Jingfukang Granule (0.5 mg · kg(-1) · d(-1)) were given by intragastric administration, once daily for 10 days. The expression levels of MMP-1, MMP-3 and TIMP-1 genes in the pulpiform nucleus of cervical intervertebral discs were detected by RT-PCR and changes of the ultrastructure of the pulpiform nucleus observed under transmission electron microscope. RESULTS: Compared to the control group, the expression levels of MMP-1 mRNA and MMP-3 mRNA of the cervical intervertebral disc tissues were significantly up-regulated in the model group (P < 0.05), and that of TIMP-1 mRNA was obviously down-regulated in the model group (P < 0.05). After the treatment, the increased expression of MMP-1 mRNA and MMP-3 mRNA and the decreased expression of TIMP-1 mRNA were reversed by acupotomy lysis and medication (P < 0.05) except TIMP-1 mRNA in the medication group (P > 0.05). No significant differences were found between the Jiaji (EX-B 2) and cervico-acupoint groups in down-regulating MMP-1 mRNA and MMP-3 mRNA expression and up-regulating TIMP-1 mRNA expression (P > 0.05). Results of electron microscope examinations showed that the ultrastructural injury changes of cells of the pulpiform nucleus were relatively milder in the Jiaji (EX-B 2) and cervico-acupoint groups, followed by the medication group in comparison with those of the model group. CONCLUSION: Acupotomy lysis at acupoints around the neck can improve the ultrastructural changes of cells of the pulpiform nucleus of cervical intervertebral discs in IVDD rats, which is possibly by regulating the expression of MMP-1, MMP-3 and TIMP-1 genes.

Structural and Ultrastructural Analysis of the Cervical Discs of Young and Elderly Humans.

Several studies describing the ultrastructure and extracellular matrix (ECM) of intervertebral discs (IVDs) involve animal models and specimens obtained from symptomatic individuals during surgery for degenerative disease or scoliosis, which may not necessarily correlate to changes secondary to normal aging in humans. These changes may also be segment-specific based on different load patterns throughout life. Our objective was to describe the ECM and collagen profile of cervical IVDs in young (G1 - <35 years) and elderly (G2 - >65 years) presumably-asymptomatic individuals. Thirty cervical discs per group were obtained during autopsies of presumably-asymptomatic individuals. IVDs were analyzed with MRI, a morphological grading scale, light microscopy, scanning electron microscopy (SEM) and immunohistochemistry (IHC) for collagen types I, II, III, IV, V, VI, IX and X. Macroscopic degenerative features such as loss of annulus-nucleus distinction and fissures were found in both groups and significantly more severe in G2 as expected. MRI could not detect all morphological changes when compared even with simple morphological inspection. The loose fibrocartilaginous G1 matrix was replaced by a denser ECM in G2 with predominantly cartilaginous characteristics, chondrocyte clusters and absent elastic fibers. SEM demonstrated persistence of an identifiable nucleus and Sharpey-type insertion of cervical annulus fibers even in highly-degenerated G2 specimens. All collagen types were detected in every disc sector except for collagen X, with the largest area stained by collagens II and IV. Collagen detection was significantly decreased in G2: although significant intradiscal differences were rare, changes may occur faster or earlier in the posterior annulus. These results demonstrate an extensive modification of the ECM with maintenance of basic ultrastructural features despite severe macroscopic degeneration. Collagen analysis supports there is not a "pathologic" collagen type and changes are generally similar throughout the disc. Understanding the collagen and ultrastructural substrate of degenerative changes in the human disc is an essential step in planning restorative therapies.

Morphological changes in disc herniation in the lower cervical spine: an ultrastructural study.

INTRODUCTION: The basis of disc degeneration is still unknown, but is believed to be a cell-mediated process. Apoptosis might play a major role in degenerative disc disease (DDD). The aim of this study was to correlate the viability of disc cells with the radiological degeneration grades (rDG) in disc herniation. MATERIALS AND METHODS: Forty anterior IVD's (C4-C7) from 39 patients with DDD were studied histologically and ultrastructurally to quantify healthy, "balloon", chondroptotic, apoptotic and necrotic cells. Patients were classified to their rDG, as having either prolapse (P: DGII + III) and/or osteochondrosis (O: DGIV + V). Similar studies were undertaken on eight control discs. RESULTS: Cell death by necrosis (mean 35%) was common but differed not significantly in both groups. All patients with a disc prolapse DGII + III revealed balloon cells (iAF: mean 32%). All appeared alive and sometimes were hypertrophic. However, significantly less balloon cells were found in the O-Group. Control samples revealed no evidence of "balloon" cells in DGII and only a minor rate in DGIII. CONCLUSION: According to the different rDG, quantitative changes were obvious in healthy and "balloon" cells, but not for cell death. At the moment it can only be hypothesized if "balloon" cells are part of a repair strategy and/or cause of disc herniation.

Accumulation of p62 in degenerated spinal cord under chronic mechanical compression: functional analysis of p62 and autophagy in hypoxic neuronal cells.

Intracellular accumulation of altered proteins, including p62 and ubiquitinated proteins, is the basis of most neurodegenerative disorders. The relationship among the accumulation of altered proteins, autophagy, and spinal cord dysfunction by cervical spondylotic myelopathy has not been clarified. We examined the expression of p62 and autophagy markers in the chronically compressed spinal cord of tiptoe-walking Yoshimura mice. In addition, we examined the expression and roles of p62 and autophagy in hypoxic neuronal cells. Western blot analysis showed the accumulation of p62, ubiquitinated proteins, and microtubule-associated protein 1 light chain 3 (LC3), an autophagic marker, in the compressed spinal cord. Immunohistochemical examinations showed that p62 accumulated in neurons, axons, astrocytes, and oligodendrocytes. Electron microscopy showed the expression of autophagy markers, including autolysosomes and autophagic vesicles, in the compressed spinal cord. These findings suggest the presence of p62 and autophagy in the degenerated compressed spinal cord. Hypoxic stress increased the expression of p62, ubiquitinated proteins, and LC3-II in neuronal cells. In addition, LC3 turnover assay and GFP-LC3 cleavage assay showed that hypoxic stress increased autophagy flux in neuronal cells. These findings suggest that hypoxic stress induces accumulation of p62 and autophagy in neuronal cells. The forced expression of p62 decreased the number of neuronal cells under hypoxic stress. These findings suggest that p62 accumulation under hypoxic stress promotes neuronal cell death. Treatment with 3-methyladenine, an autophagy inhibitor decreased the number of neuronal cells, whereas lithium chloride, an autophagy inducer increased the number of cells under hypoxic stress. These findings suggest that autophagy promotes neuronal cell survival under hypoxic stress. Our findings suggest that pharmacological inducers of autophagy may be useful for treating cervical spondylotic myelopathy patients.

Variant inferior root of ansa cervicalis / Variación de la raíz inferior del asa cervical

Ansa cervicalis is a loop of nerves found in the anterior wall of the carotid sheath in the carotid triangle of neck innervating infrahyoid muscles. Its superior root has fibres from the first cervical nerve that leaves the hypoglossal nerve and joins the inferior root formed by the branches from the second and third cervical nerves. The two roots join to form ansa cervicalis. The ansa cervicalis nerve formation is relatively complex, as its course and location along the great vessels of the neck vary. In the present case on the left side of an adult male cadaver the inferior root was absent and the contributions from C2 and C3 were joining independently with the superior root of ansa to form ansa cervicalis. However no such variation was found in the ansa cervicalis formation on the right side.

El asa cervical es un loop de nervios que se encuentra en la pared anterior de la vaina carotídea, en el triángulo carotídeo del cuello, que inerva los músculos infrahioideos. Su raíz superior tiene fibras del primer nervio cervical que sale del nervio hipogloso y se une a la raíz inferior formada por las ramas de los nervios cervicales segundo y tercero. Las dos raíces se unen para formar el asa cervical. La formación del asa cervical del nervio es relativamente compleja, ya que su curso y ubicación varía a lo largo de los grandes vasos del cuello. En el lado izquierdo del cuello de un cadáver adulto de sexo masculino, la raíz inferior del asa cervical estaba ausente y las contribuciones de los ramos de C2 y C3 se unieron, de forma independiente, con la raíz superior del asa y así formar el loop.

Posterior osseous bridging of C1.

The sulcus of the vertebral artery is located behind the lateral mass of the atlas and in some cases is converted into a foramen by anomalous ossification known as the posterior ponticulus (osseous bridge). This study involved anatomical observations of 158 isolated anatomical specimens of dry C1 vertebrae. The incidence and types of posterior osseous bridging were identified for the 158 dry samples of atlas vertebrae. In nine (5.6%) dry C1 vertebrae, partial osseous bridging was detected (bilaterally in eight vertebrae and unilaterally on the left in one). Complete osseous bridging (arcuate foramen) was observed in six (3.8%) dry C1 vertebrae (bilaterally in one vertebra, unilaterally on the left in three, and on the right in two). Awareness of the types of posterior osseous bridging of C1 in craniocervical junction surgery is essential, and may be helpful in surgical interventions in this region.

An ultrastructural study on the ligamentum flavum of the cervical spine in patients with ossification of the posterior longitudinal ligament.

Some histological analyses of the ossification of the posterior longitudinal ligament (OPLL) have been reported, but no ultrastructural studies of the ligamentum flavum (LF) in patients with OPLL have been published to date. To understand the pathology of the ossification of the spinal ligament, we examined, by electron microscopy, ultrastructural changes in the LF in cases of OPLL and made a comparison with the LF in cervical spondylotic myelopathy (CSM). Subjects were three men and two women with cervical OPLL who underwent longitudinal spinous process-splitting laminoplasty. During surgery, a small piece of the LF was collected from C2-C3 to C7-T1 and was then analyzed by light and electron microscopy. We observed atrophic elastic bundles with a two-layer structure and disarrangement, a partially torn area, the disappearance of microfibrils, and an enlarged interstitium with an irregular alignment of collagen fibrils. We observed some properties of a cell preceding its death: the initial phase may be the disappearance of the plasma-membrane, followed by the scattering of many organellae around its degenerated nucleus. Finally, many extracellular plasma membrane-invested particles that resemble matrix vesicles remain there without phagocytosis. These results suggest that ultrastructural abnormalities exist in the spinal ligament in cases of ossification of the spinal ligament.

[Noninvasive, accurate and reliable measurement of cervical spine motion with a 3D real-time ultrasound motion analyzer]. / Nichtinvasive, akkurate und reliable Messung der Halswirbelsäulenbeweglichkeit mittels ultraschallgestützter 3D-Echtzeit-Bewegungsanalyse.

AIMS: The kinematic analysis of cervical spine motion is important to assess objectively the effects of therapeutic interventions. In this study, precision and reliability of a 3D ultrasound motion analyser was determined. Using this tool the physiologic range of movement of healthy volunteers was assessed. The aim was to test the clinical practicability of this system. METHODS: The active and passive cervical spine range of motion of 20 healthy volunteers with a mean age of 23 years (range 19-28 years) was determined using a CMS 3D ultrasound realtime motion analyser (Zebris Medizintechnik, Tübingen, Germany). Precision was assessed by comparison with a precision goniometer. Two observers determined the inter-rater and retest reliability and the Pearson correlation coefficients were calculated. RESULTS: The maximum measurement difference between CMS and precision goniometer was 0.6 degree. Inter-rater- and retest reliability correlated significantly (0.84 < r < 0.96, p < 0.001). The range of motion found by the CMS corresponded well with motion values determined using other devices. CONCLUSIONS: The range of motion of the cervical spine can be assessed accurately and reliably using a 3D ultrasound motion analyser. The CMS motion analyser is suitable for clinical practice.

Uncinate processes and Luschka joints influence the biomechanics of the cervical spine: quantification using a finite element model of the C5-C6 segment.

A fully three-dimensional finite element model of a C5-C6 motion segment of the human spine was developed and validated for the purpose of investigating the biomechanical significance of uncinate processes and Luschka joints. The original intact cervical model was modified to create two additional models. The first simulated the absence of Luschka joints by replacing the fissures with continuous annulus fibrosus and leaving the uncinate processes intact. The second model simulated a surgical resection of the uncinate processes, while leaving the Luschka joints intact. The results of these two models were compared with the intact model, which served as a baseline; thus, the relative contributions of these two structures to cervical motion were established. With use of our model, it was possible, for the first time, to provide quantitative data concerning the source of coupled motions in the lower cervical spine. In principle, the results from this model support the hypothesis of Penning and Wilmink. Our results indicate that the facet joints and Luschka joints are the major contributors to coupled motion in the lower cervical spine and that the uncinate processes effectively reduce motion coupling and primary cervical motion (motion in the same direction as load application), especially in response to axial rotation and lateral bending loads. Luschka joints appear to increase primary cervical motion, showing an effect on cervical motion opposite to that of the uncinate processes. Surgeons should be aware of the increase in motion accompanied by resection of the uncinate processes.

Histiocytic necrotizing lymphadenitis (Kikuchi's disease) of the cervical lymph nodes.

Histiocytic necrotizing lymphadenitis (HNL), or Kikuchi's disease, is a benign cause of lymph node enlargement of unknown origin. It may be mistaken for malignant lymphoma, both clinically and histologically. Though well recognized in the pathological literature few clinicians are aware of the disease. We present a case of cervical HNL and review the literature.

Anatomic relationships of the cervicothoracic junction.

STUDY DESIGN: This study analyzed the anatomic relationships between bony structures and soft tissues of the cervicothoracic junction. OBJECTIVES: To provide composite reference data for intrasegmental and intersegmental gradients of anatomic variation within the cervical-thoracic junction. SUMMARY OF BACKGROUND DATA: Because the risk of soft tissue damage during posterior spinal stabilization, an understanding of bony and soft tissue changes in the cervicothoracic junction is necessary. METHODS: Three-hundred-twenty-four cross-sectional spinal segments from nine spines were analyzed to characterize cervicothoracic junctional anatomy. RESULTS: There were predictable cranial-to-caudal alterations in both bone and soft tissue anatomy of the cervicothoracic junction. Neural and vascular structures directly anterior to the lateral mass or transverse process and lateral to the pedicle tend to decrease in frequency, whereas measured parameters of the vertebrae increase in size from C5-T3, except for pedicle dimensions that tend to increase at the C7-T1 junction. CONCLUSION: The anatomic changes that occur within the cervicothoracic junction are consistent and predictable, and their recognition should lead to a better appreciation of their clinical implications.

Intervertebral variation in trabecular microarchitecture throughout the normal spine in relation to age.

The vertebral bodies of the complete spine (C-3-L-5) were removed in 26 autopsy cases without evidence for primary or secondary bone disease (13 males aged 19-79 years and 13 females aged 17-90 years). A sagittal segment through the center of all vertebral bodies was embedded undecalcified in hydroxyethylmethacrylate and processed to so-called surface stained block grindings. Histomorphometric analysis of the complete segment was performed using a computer-assisted image analysis system (IBAS 2000). The structural parameters investigated were bone volume (BV/TV) and trabecular interconnection quantificated by trabecular bone pattern factor (TBPf). A close correlation of BV/TV and TBPf was found in all vertebral bodies irrespective of vertebral region (r = 0.8, p < 0.001). This indicates that the age-related decrease of trabecular bone mass is primarily the consequence of the transformation from plates to rods and the loss of whole trabecular structures. This basic principle is valid throughout the complete spine. However, the systematic analysis of vertebral trabecular bone from C-3 to L-5 revealed a significant intervertebral variation of trabecular microarchitecture. The density of trabecular structure of cervical vertebrae is much higher than that of thoracic and lumbar vertebrae (p < 0.001). The extent of age-related loss of trabecular bone mass and structure showed a decrease within the spine from the caudal to the cranial region (p < 0.05). The loss of bone volume in individuals between the ages of 30 and 80 years in the lumbar spine was 53%, whereas in the thoracic spine the decrease was 41%, and in the cervical spine only 24%.(ABSTRACT TRUNCATED AT 250 WORDS)

[Angio-fibrolipoma in epidural space of the vertebral canal]. / Naczyniako-wlókniako-tluszczak w przestrzeni zewnatrzoponowej kanalu kregowego.

55-yera old patient with angio-fibro-lipoma of thoracic part of the vertebral canal was described. This rare, benign tumour was totally removed. Subsequently the neurological state of the patient improved.

[Malherbe's calcifying epithelioma (pilomatrixoma) of the cervical middle line. A case report]. / Epitelioma calcificante de malherbe (pilomatrixoma) en línea media cervical. Presentación de un caso.

Calcifying epithelioma of Malherbe (Pilomatrixoma) is a rare benign tumor, starting in the hair matrix and clinically resembling a hard subcutaneous swelling. The most common sites of localization are the head and neck regions. Women show a sizeable incidence. We report a case located in the prelaryngeal area and emphasize cytodiagnostic by fine needle aspiration. The correct treatment consists in pure and simply surgical excision under local anesthesia.

[Lateral isolated neurofibroma of the neck]. / Neurofibroma solitario del espacio laterocervical.

The neurofibroma (NF) of the neck is a neurogenic tumor seldom presented. Sometimes solitary, as the case reported, or within the context of von Recklinghausen's disease. When diagnosis is doubtful either the TAC, the RMN of the PAAF show its usefulness. However the ultimate diagnosis is made through the histologic exam of the removed lump, which is the elective treatment.

Numbers and proportions of unmyelinated axons at cervical levels in the fasciculus gracilis of monkey and cat.

The present study is a quantitative analysis of the unmyelinated fiber population in the fasciculus gracilis of the second cervical segment of cat and monkey. We find that unmyelinated fibers represent 13.7% of the total fiber population in this pathway in the cat and 18.9% in the monkey (Macaca fascicularis). The existence of such large numbers of these axons suggests that there may be a sizeable ascending fine primary afferent pathway in the fasciculus gracilis in cat and monkey whose destination is presumably the dorsal column nuclei. These findings are of interest in regard to classic ideas that the afferent fibers in the dorsal columns are large myelinated fibers that convey fine discriminative information to the dorsal column nuclei.

Induction and characterization of an interface tissue by implantation of methylmethacrylate cement into the posterior part of the cervical spine of the dog.

After the implantation of methylmethacrylate cement into the posterior part of the cervical spine of the dog, a thick layer of connective tissue forms at the bone-cement interface. The tissue is six to eight millimeters thick and in all animals it surrounds the dorsal and lateral aspects of the masses of implanted cement, grows between the undersurface of the cement and the bone of the posterior elements, and completely covers that bone. This tissue was examined by light and electron microscopy and its collagenous components were extracted and analyzed biochemically by gel electrophoresis. Specific extracellular matrix proteins in the tissue at the bone-cement interface were also localized by immunohistochemistry. The tissue at the host-cement interface contained zones of fibrocytes and plump and teardrop-shaped cells within a collagenous matrix. Type-I, Type-III, and Type-V collagen were extracted and were identified by gel electrophoresis. Type-V collagen and fibronectin were localized predominantly around the plump and teardrop-shaped cells. Type-IV collagen and laminin were localized predominantly in an area just beneath the teardrop-shaped cells at the surface of the tissue overlying the cement, suggesting that a basement-membrane-like tissue had formed in this area.

Ultrastructural study of calcification process in the ligamentum flavum of the cervical spine.

The ultrastructure of the formation of calcified nodules in three cases of symptomatic calcification of the cervical ligamentum flavum were studied. In some areas of the ligament, extracellular plasma membrane-invested matrix vesicles and thick wall-bound matrix giant bodies with or without mineralized deposits were present. These calcified vesicles and bodies were also encountered in the wide mineralized areas among the collagen and elastic fibers in the ligament. The mineralization process of the calcified nodules in the ligamentum flavum implies that matrix vesicles and matrix giant bodies acquire mineralized precipitates; then some gather in clusters. Calcified deposits may spread to collagen and elastic fibers contiguous with the calcified vesicles and bodies, and some eventually coalesce to make a calcified nodule.

Septal afferents to the area dentata terminate on vasoactive intestinal polypeptide (VIP)-like immunoreactive, non-pyramidal neurons. An electron-microscopic immunocytochemical degeneration study in the rat.

Thermic lesions in the medial septum of adult rats result in dark degeneration of terminal boutons in the stratum moleculare and hilus of the area dentata. While most of the degenerating terminals are in synaptic contact with non-reactive cells, part of them end on dendrites of VIP-like immunoreactive neurons.