Monkey flexor and abductor pollicis brevis motoneuron pools: Proximal dendritic trees and small motoneurons.

Transverse sections of the monkey cervical spinal cord from a previous study (Jenny and Inukai, 1983 [1]) were reanalyzed using Neurolucida to create a three-dimensional display of flexor pollicis brevis and abductor pollicis brevis (FAbPBr) motoneurons and dendrites that had been jointly labeled with horse radish peroxidase (HRP). These data were correlated with similar data from a reanalysis of an extensor digitorum communis (EDC) motoneuron pool (Jenny, Cheney, and Jenny, 2018 [2]). The FAbPBr motoneuron columns were located in the C8 (caudal) and T1 segments of the spinal cord and within the most dorsal and medial regions of the motor column pools that innervate hand muscles. Small motoneurons (cell body areas less than 500 µm2 and presumed to be gamma motoneurons) comprised about four percent of the motoneurons and were located throughout the length of the motoneuron pool. HRP labeled dendrites extended radially (360°) from the motoneuron soma but greater numbers of dendrites were directed either dorsomedial to the base of the dorsal horn or medial to the ventromedial gray matter. The longer HRP labeled dendrites and their branch dendrites usually continued in the same radial direction as when originating from the cell body or proximal dendrite. As such we considered the radial direction of the longer HRP labeled dendrites to be a reasonable estimate of the radial direction of the more distal dendritic trees [2]. Both the EDC and FAbPBr motoneuron groups had a greater number of dendrites oriented in dorsal and medial directions from the motoneuron column. Our data continue to suggest that motoneuron dendritic trees have direction-oriented dendrites that extend toward functional terminal regions.

Apoptosis of endplate chondrocytes in cervical kyphosis is associated with chronic forward flexed neck: an in vivo rat bipedal walking model.

BACKGROUND: This study was undertaken to establish a rat bipedal walking model of cervical kyphosis (CK) associated with chronic forward flexed neck and assess the effects of chronic forward flexed neck on endplate chondrocytes. METHODS: Forty-eight 1-month-old Sprague-Dawley rats were randomly divided into 3 groups: forward flexed neck group (n = 16), bipedal group (n = 16), and normal group (n = 16). Cervical curves were analyzed on a lateral cervical spine X-ray using Harrison's posterior tangent method before the experiment and at 2-week intervals for a 6-week period. Histologic changes in cartilaginous endplate chondrocytes were observed using hematoxylin and eosin (H&E) staining, transmission electron microscopy (TEM), and terminal deoxyribonucleotidyl transferase (TdT)-mediated dUTP nick-end labeling. RESULTS: Radiographic findings suggested a significantly decreased cervical physiological curvature in the forward flexed neck group over the 6-week follow-up; normal cervical curves were maintained in other groups. The average cervical curvature (C2-C7) was - 7.6 ± 0.9° in the forward flexed neck group before the experiment, - 3.9 ± 0.8° at 2 weeks post-experiment, 10.7 ± 1.0° at 4 weeks post-experiment, and 20.5 ± 2.1° at the last follow-up post-experiment. Histologically, results of H&E staining unveiled that cartilaginous endplate chondrocytes were arranged in an irregular fashion, with the decreased number at the observation period; the incidence of apoptotic cells in the forward flexed neck group was noticeably higher at the 6-week follow-up than that in other groups. CONCLUSIONS: CK developed as the result of chronic forward flexed neck. Histologic changes suggested that chondrocyte apoptosis may play a critical role in the development of cervical kyphotic deformity associated with chronic forward flexed neck.

Visual information increases the indirect corticospinal excitation via cervical interneurons in humans.

Modulatory actions of inputs from the visual system to cervical interneurons (IN) for arm muscle control are poorly understood in humans. In the present study, we examined whether visual stimulation modulates the excitation of cervical IN systems mediating corticospinal tract (CST) inputs to biceps brachii (BB). Twenty-eight healthy volunteers were seated, and electromyogram recordings from the BB were performed across six experiments, each with discrete objectives. A flash stimulator for visual stimulation (50-µs duration) was placed 60 cm from the participant's eye. The CST was stimulated with transcranial magnetic/electrical stimulation (TMS/TES, respectively) contralateral to the recording site. Visual stimulation with TMS/TES was randomly delivered during weak tonic BB contractions. Single TMS/TES-induced motor-evoked potentials (MEPs) were markedly enhanced from 60-100 ms after visual stimulation compared with the control condition. The MEPs were significantly increased by combining the electrical stimulation of the ulnar nerve at the wrist [7.5-12 ms of nerve stimulation (NERVE)/TMS interval] with and without visual stimulation compared with the algebraic summation of responses obtained with either TMS or NERVE. Interestingly, the combined stimulation-induced MEP facilitation was significantly increased after visual stimulation compared with the control. Single motor unit (MU) recording also revealed the further enhancement of combined stimulation effects on the firing probabilities of MU during visual stimulation, which was observed in the peaks of the peristimulus time histogram, 1-2 ms later than the onset latency. The present findings suggest that visual stimulation facilitates the oligosynaptic CST excitation of arm motoneurons mediated by the cervical IN system.NEW & NOTEWORTHY To date, little is known about how visual information modulates the human cervical motor systems, including the presumed interneuron (IN) circuitry. This study demonstrates that photic visual stimulation influences presumed oligosynaptic corticospinal transmission to arm motoneurons, which are mediated by cervical INs. In animals, these systems are known to be crucial for visually guided switching movements, and similar visual input systems to INs may exist in humans.

Short-Term Environmental Enrichment is a Stronger Modulator of Brain Glial Cells and Cervical Lymph Node T Cell Subtypes than Exercise or Combined Exercise and Enrichment.

Physical exercise (PE) and environmental enrichment (EE) can modulate immunity. However, the differential effects of short-term PE, EE, and PE + EE on neuroimmune mechanisms during normal aging has not been elucidated. Hence, a cohort of 3-, 8-, and 13-month-old immunologically unchallenged C57BL/6 wild-type mice were randomly assigned to either Control, PE, EE, or PE + EE groups and provided with either no treatment, a running wheel, a variety of plastic and wooden objects alone or in combination with a running wheel for seven weeks, respectively. Immunohistochemistry and 8-color flow cytometry were used to determine the numbers of dentate gyrus glial cells, and the proportions of CD4+ and CD8+ T cell numbers and their subsets from cervical lymph nodes, respectively. An increase in the number of IBA1+ microglia in the dentate gyrus at 5 and 10 months was observed after EE, while PE and PE + EE increased it only at 10 months. No change in astroglia number in comparison to controls were observed in any of the treatment groups. Also, all treatments induced significant differences in the proportion of specific T cell subsets, i.e., CD4+ and CD8+ T naïve (TN), central memory (TCM), and effector memory (TEM) cells. Our results suggest that in the short-term, EE is a stronger modulator of microglial and peripheral T cell subset numbers than PE and PE + EE, and the combination of short-term PE and EE has no additive effects.

Autologous Bone Harvest in Anterior Cervical Spine Surgery: A Quantitative and Qualitative In Vitro Analysis of Cadaveric Tissue.

BACKGROUND: The cervical spine may be used as a harvesting site of local autograft material during anterior cervical discectomy and fusion procedures. We analyzed the quality and composition of bone grafts obtained from different parts of the cervical vertebrae in a cadaveric model. METHODS: Five fresh adult human cadavers with intact cervical spines were used. Using a Smith-Robinson anterior approach to expose C4-5 and C5-6 vertebrae, samples from 4 vertebral sites were harvested under a microscope. Anterior osteophytes were removed piecemeal by a Leksell rongeur (sample A). A high-speed burr was used to drill the endplates of C4-5 and C5-6 (sample C) and uncovertebral joints of C4-5 (sample B) and C5-6 (sample D). Then 20 slides (4 per cadaver) were prepared and analyzed. RESULTS: Tissue fragmentation was associated with use of the high-speed burr. Sample A had minimal tissue fragmentation. Samples B-D showed moderate to high fragmentation. Cartilage was found in all samples. Of the 20 slides, 6 contained soft tissues (sample A in 4, sample D in 2). Disc material was identified in 6 slides (sample A in 1, sample B in 4, sample D in 1). Sample A had the greatest number of intact osteocytes and chondrocytes, and sample B had the least. CONCLUSIONS: Anterior osteophytes provide the highest number of osteocytes, with the highest osteocyte/chondrocyte ratio. Osteocyte viability is a function of vertebral body site and collection technique, with fragmentation caused by use of a high-speed burr decreasing the number of viable osteocytes.

JNK phosphorylation promotes natural degeneration of cervical endplate chondrocytes by down-regulating expression of ANK.

BACKGROUND: Endplate degeneration leads to accelerated degeneration of the intervertebral disc. The importance of endplate chondrocytes in this process is unclear. Many cellular processes in chondrocytes are controlled by activated c-Jun N-terminal kinases (JNK) and protein kinase B (AKT). However, the involvement of their pathways in the degeneration process needs to be elucidated. AIM: To study activation of JNK and AKT signaling pathways and their significance for degeneration of endplate chondrocytes, as well as involvement of progressive ankylosis protein (ANK) in this process. MATERIALS AND METHODS: Rat primary chondrocytes were grown to confluence and subcultured until passage 4. Morphological appearances (microscope, hematoxylin & eosin staining, toluidine blue staining) and proliferation rates of cells (MTT test) were observed. Further, levels of type II collagen, aggrecan, phosphorylated JNK and AKT, total JNK, AKT and ANK were evaluated by qPCR, flow cytometry and Western blot assays. Furthermore, inhibition experiments with SP600125, the JNK inhibitor, were carried out in the passage 4 cells to assess the effects of the JNK pathway on natural degeneration of endplate chondrocytes. RESULTS: The proliferative speed of endplate chondrocytes progressively decreased during passaging. Expressions of type II collagen and aggrecan were significantly decreased with cells at higher passages. Furthermore, phosphorylation of JNK, but not AKT, was significantly up-regulated and accompanied by reduced ANK expression. Inhibition of the JNK pathway increased expression of type II collagen, aggrecan and ANK and facilitated proliferation rates. CONCLUSIONS: Phosphorylation of JNK promotes natural degeneration of cervical endplate chondrocytes, likely by down-regulating ANK expression.

[Significance and expression of Sirt1 gene in human cervical endplate chondrocyte in vitro degeneration].

OBJECTIVE: To observe the expression changes of Sirt1 gene and examine the role and significance of degenerative process in human cervical endplate chondrocytes through a degeneration model of human cervical vertebral endplate chondrocyte. METHODS: Cartilage endplates of 30 patients were divided into control group (n = 16) with cervical vertebral fracture or dislocation and cervical spondylosis group (n = 14) with cervical spondylotic myelopathy. Endplate chondrocytes were isolated by enzyme digestion and cultured in vitro for 10 days. The differences of endplate chondrocytes from normal and degenerative cartilage endplates were observed by inverted phase-contrast microscope, hematoxylin and eosin staining and toluidine blue staining. Real-time reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were used to detect the mRNA expressions of Sirt1, collagen II and aggrecan. RESULTS: Compared with the normal group, the cellular morphology of degenerative group showed spindle-shaped changes. The mRNA expression of Sirt1 (P = 0.034) significantly decreased. Aggrecan (P = 0.0063) and collagen II (P = 0.0072) decreased also markedly. CONCLUSION: Sirt1 gene expression is significantly down-regulated in degenerative human cervical endplate chondrocytes. Regulating the expression of Sirt1 gene may block or delay the occurrence of human cervical endplate cartilage degeneration.

Increased number of neurons in the cervical spinal cord of aged female rats.

In the brain, specific signaling pathways localized in highly organized regions called niches allow the persistence of a pool of stem and progenitor cells that generate new neurons in adulthood. Much less is known about the spinal cord where a sustained adult neurogenesis is not observed. Moreover, there is scarce information concerning cell proliferation in the adult mammalian spinal cord and virtually none in aging animals or humans. We performed a comparative morphometric and immunofluorescence study of the entire cervical region (C1-C8) in young (5 mo.) and aged (30 mo.) female rats. Serum prolactin (PRL), a neurogenic hormone, was also measured. Gross anatomy showed a significant age-related increase in size of all of the cervical segments. Morphometric analysis of cresyl violet stained segments also showed a significant increase in the area occupied by the gray matter of some cervical segments of aged rats. The most interesting finding was that both the total area occupied by neurons and the number of neurons increased significantly with age, the latter increase ranging from 16% (C6) to 34% (C2). Taking the total number of cervical neurons the age-related increase ranged from 19% (C6) to 51% (C3), C3 being the segment that grew most in length in the aged animals. Some bromodeoxyuridine positive-neuron specific enolase negative (BrdU(+)-NSE(-)) cells were observed and, occasionally, double positive (BrdU(+)-NSE(+)) cells were detected in some cervical segments of both young and aged rats groups. As expected, serum PRL increased markedly with age. We propose that in the cervical spinal cord of female rats, both maturation of pre-existing neuroblasts and/or possible neurogenesis occur during the entire life span, in a process in which PRL may play a role.

[Morphologic and apoptotic changes of cervical posterior longitudinal ligament fibroblasts in response to in vitro mechanical stretch].

OBJECTIVE: To investigate the effect of mechanical stretch force on the morphologic and apoptotic changes of fibroblasts derived from the OPLL (ossification of posterior longitudinal ligaments) patients. METHODS: The third passage cells were collected and subjected to 10% elongations cyclic mechanical stretching for 6 h and 24 h with Flexercell 4000(TM) strain unit. Their morphologic changes were observed and the post-stretching apoptotic rates quantified by flow cytometer. The gene expressions of alkaline phosphatase (ALP), collagen types I (COL I) and osteocalcin (OC) were examined. RESULTS: The treated cells were arranged along the vertical direction of force. Stretch force led to a slight increase of apoptosis rate at 6 h and a significant increase of apoptosis rate at 24 h. No significant difference in cellular senescence was observed between control group and treated group. The mRNA expressions of ALP, COL I and OC were positively up-regulated by cyclic stretch at 24 h. CONCLUSION: Stretching force can affect the cellular morphology, promote the osteogenic differentiation and enhance the cellular apoptosis.

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.

A comparison of mesenchymal precursor cells and amnion epithelial cells for enhancing cervical interbody fusion in an ovine model.

BACKGROUND: Rapid, reliable fusion is the goal in anterior cervical diskectomy and fusion. Iliac crest autograft has a high rate of donor-site morbidity. Alternatives such as bone graft substitutes lack osteoinductivity, and recombinant bone morphogenetic proteins risk life-threatening complications. Both allogeneic mesenchymal precursor cells (MPCs) and amnion derived epithelial cells (AECs) have osteogenic potential. OBJECTIVE: To compare for the first time the capacity of MPCs and AECs to promote osteogenesis in an ovine model. METHODS: Five groups of 2-year-old ewes were subjected to C3-4 anterior cervical diskectomy and fusion with a Fidji interbody cage packed with iliac crest autograft alone (group A; n = 6), hydroxyapatite-tricalcium phosphate Mastergraft granules (HA/TCP) alone (group B; n = 6), HA/TCP containing 5 million MPCs (group C; n = 6), or HA/TCP containing 5 million AECs (group D; n = 5); group E was made up of age-matched nonoperative controls (n = 6). At 3 months, animals were euthanized and quantitative multislice computed tomography, functional radiography, biomechanics, histology, and histomorphometry were performed. RESULTS: No procedure- or cell-related adverse events were observed. There was significantly more fusion in the MPC group (C) than in group A, B, or D. Computed tomography scan at 3 months revealed that 5 of 6 MPC-treated animals (83%) had continuous bony bridging compared with 0 of 5 AEC-treated and only 1 of 6 autograft- and 2 of 6 HA/TCP-treated animals (P = .01). CONCLUSION: Implantation of allogeneic MPCs in combination with HA/TCP within an interbody spacer facilitates interbody fusion after diskectomy. The earlier, more robust fusion observed with MPCs relative to autograft and HA/TCP bone substitute indicates that this approach may offer a therapeutic benefit.

Immunohistochemical and histological study of human uncovertebral joints: a preliminary investigation.

STUDY DESIGN: A descriptive cadaveric study. OBJECTIVE: To investigate the anatomy and innervation of the uncovertebral joint to determine if it is synovial in nature and capable of generating pain. SUMMARY OF BACKGROUND DATA: There is controversy with regard to the anatomic and histological makeup of the uncovertebral interface with some authors considering it a joint and others disc tissue. No research has investigated the presence of pain generating neurotransmitters within the uncovertebral cartilaginous and capsular tissue. METHODS: Tissue from uncovertebral capsule and cartilage was harvested for each uncovertebral surface starting at the C2-C3 to the C6-C7 cervical segment. The tissue was placed in 4% paraformaldehyde fixative, then dehydrated and embedded in paraffin. Ten micron sections were cut through the tissue blocks and mounted on slides. The tissue was rehydrated and either stained with hematoxylin and eosin (H and E) or immunostained with antisera against protein gene product 9.5 (PGP 9.5), substance P (SP), neuropeptide Y (NPY), and calcitonin gene-related peptide (CGRP). RESULTS: The sample consisted of 2 unembalmed fresh male human cadavers of a mean age of 83 years. Chondrocytes and synoviocytes were identified at the capsular tissue of each uncovertebral interface from C2-C3-C6-C7. Immunoreactivity for PGP 9.5, SP, CGRP, and NPY was observed at all uncovertebral interface levels in capsular tissue. CONCLUSION: The presence of both synoviocytes and chondrocytes has been recorded in the present study, suggesting that the uncovertebral interface is synovial in nature. Immunoreactivity to PGP 9.5, SP, CGRP, and NPY indicates the presence of nerve fibers from both the somatic and autonomic nervous systems. These findings suggest that the uncovertebral joints are potential pain generators in the cervical spine.

A novel method to evaluate vertebral remodeling by radiography following anterior cervical decompression and interbody fixation with cylindrical cages: a contrast-comparing method using "Scion image".

In an attempt to study bone remodeling by noninvasive methods, spinal bone radiodensity was assessed in five patients treated with anterior cervical decompression and fusion (ACDF) using cylindrical titanium cages. Plain radiographs were used to study specific areas of vertebral bone interposed in two-level cages with the two cephalad vertebrae for controls. Measurements were made immediately after surgery and 1, 3, 6, 12 and 18 months postoperatively. The data were analyzed quantitatively with a contrast-comparing method (CCM) using "Scion image". There were two cyclical changes in vertebral remodeling. First, in all patients there were gradual increases in bone density at the ventral part compared to the dorsal part of the vertebral body for up to 12 months; then the density decreased at 18 months. Second, a linear gradient in radiodensity from the ventral part to the dorsal part of the vertebral body observed immediately following spinal fusion gradually disappeared by 12 months; nonhomogeneous distributions of trabecular bone were appeared. Then, the linear gradient in density appeared again at 18 months. This investigation helps elucidate the radiographic evidence for the remodeling of vertebral bone in patients treated with ACDF.

Morphological, stereological and radiological changes in pinealectomized chicken cervical vertebrae.

Considerable evidence supports the hypothesis that melatonin plays an important role in osteogenesis. We carried out an experiment to investigate histological and radiological changes in the cervical vertebrae of pinealectomized chickens. Thirty new-hatched chicks were divided into two groups: pinealectomized group (n=15) and nonpinealectomized control (n=15). Surgical pinealectomies were performed in Hybro broiler chickens at the age of 3 days. At 8 wk, one animal from each group was examined using computed tomography scanner to obtain density histograms of four consecutive vertebrae between the 4th and 7th cervical vertebral segments of chickens. At the end of the study, all animals were killed for subsequent measurement of bone mineral density (BMD) and macroscopic, radiographic and histopathological evaluation of specimens. We found that the profile of the density histogram in nonpinealectomized control group showed a spike profile for the vertebral body, indicating an increase in the amount of higher density tissues in this region, while a plateau-shaped profile was obtained for the vertebral body in pinealectomy group, indicating the presence of heterogenous bone tissue. Accordingly, the mean value of BMD in pinealectomy group was significantly lower at the vertebral body in chickens compared with control subjects (P<0.001). At the end of the experiment, almost all of the chickens in pinealectomy group developed a scoliotic curvature and the mean weight and length of the cervical vertebral bodies of the pinealectomized chickens were significantly lower than those of control group (P<0.001). Although the numerical density of osteocytes and osteoblasts in pinealectomy group was significantly higher than that from the control group, total number of osteocytes but not osteoblasts in cervical vetrebrae from pinealectomized animals was significantly lower than that from nonpinealectomized control animals (P<0.05). In conclusion, the results of the current study demonstrated for the first time pinealectomy-induced histomorphometrical changes in chicken vertebral column using stereological methods, suggesting that pineal gland/melatonin may have an osteoinductive effect on bone formation, but further studies are needed to elucidate the relationship of our findings with some disorders such as postmenopausal and/or senile osteoporosis.

Morphometry of cervical segments grey matter in the male rat spinal cord.

The cervical portion of the spinal cord is an area frequently affected by alterations of medical and veterinary importance. Since there is scarce quantitative anatomical data on this region, we undertook a morphometric study of the grey matter of all segments of the rat cervical spinal cord of male rats in order to generate reference patterns to be used in future experimental studies. Using image analysis software, the total spinal cord length and grey and white matter area of each segment was recorded. The morphometric characteristics of the neurones populating the laminae of the grey matter of the cervical segments was also recorded. Neurones were classified into small, medium and large sizes for each lamina and statistically compared. The present data fill an anatomical information gap by providing quantitative data about the normal anatomical features of the rat cervical cord. The anatomical data found could be used to better understand the physiological relevance of that region in the rat.

Lateral cervical nucleus projections to periaqueductal gray matter in cat.

The midbrain periaqueductal gray matter (PAG) integrates the basic responses necessary for survival of individuals and species. Examples are defense behaviors such as fight, flight, and freezing, but also sexual behavior, vocalization, and micturition. To control these behaviors the PAG depends on strong input from more rostrally located limbic structures, as well as from afferent input from the lower brainstem and spinal cord. Mouton and Holstege (2000, J Comp Neurol 428:389-410) showed that there exist at least five different groups of spino-PAG neurons, each of which is thought to subserve a specific function. The lateral cervical nucleus (LCN) in the upper cervical cord is not among these five groups. The LCN relays information from hair receptors and noxious information and projects strongly to the contralateral ventroposterior and posterior regions of thalamus and to intermediate and deep tectal layers. The question is whether the LCN also projects to the PAG. The present study in cat, using retrograde and anterograde tracing techniques, showed that neurons located in the lateral two-thirds of the LCN send fibers to the lateral part of the PAG, predominantly at rostrocaudal levels A0.6-P0.2. This part of the PAG is known to be involved in flight behavior. A concept is put forward according to which the LCN-PAG pathway alerts the animal about the presence of cutaneous stimuli that might represent danger, necessitating flight. J. Comp. Neurol. 471:434-445, 2004.

Morphology and differentiation of radial glia in the developing rat spinal cord.

Important events underlying the proper functioning of the central nervous system (CNS) include the production, assembly, and differentiation of appropriate types and numbers of cells during development. The mechanisms that control these events are difficult to unravel because of displacement of cells from their sites of origin to their permanent locations and because of the diverse cellular composition of the CNS. As in other regions of the mammalian CNS, the two major classes of neuroglial cells in the rat spinal cord are oligodendrocytes and astrocytes. In the developing spinal cord, radial glia are prominent. In this study, radial glia in the cervical region of the spinal cord were analysed. 1,1'Dioctadecyl-3,3,3'-tetramethylindocarbocyanine perchlorate (DiI) was used to determine the morphology and distribution of radial glia during spinal cord development. The DiI labelling technique enabled locating glial precursor cells during spinal cord development. Radial fibres that extended from the central canal to the pial surface were present at embryonic days 14, 16, and 18 in the developing spinal cord. Their distribution was restricted with increasing development, and by embryonic day 20 the only remaining evidence of radial glia were short radial processes in the white matter.

Anterior cervical interbody fusion with rhBMP-2 and tantalum in a goat model.

BACKGROUND CONTEXT: Tricortical autogenous iliac crest has long served as the gold standard for arthrodesis after cervical discectomy. The added morbidity resulting from bone graft harvest may be eliminated by the use of a biocompatible synthetic bone graft substitute with osteoconductive abilities, and when used with an osteoinductive agent, such as recombinant bone morphogenic protein (rhBMP)-2, it may facilitate arthrodesis similar to autograft. PURPOSE: To determine by radiographic and histologic analysis whether tantalum with and without rhBMP-2 can facilitate bony ingrowth and arthrodesis in an animal model. STUDY DESIGN/SETTING: Single-level anterior cervical discectomy and fusion was performed using a tantalum bone graft substitute with and without rhBMP-2 in a previously established goat model for anterior cervical fusion. METHODS: Eight goats underwent single-level anterior cervical discectomy and stabilization with a porous tantalum implant. There were four goats in each experimental group. Group A underwent anterior cervical stabilization with tantalum alone, whereas in Group B rhBMP-2 was added to the tantalum implant. The goats were sacrificed at 12 weeks, and their cervical spines were removed for histologic and radiological analysis. RESULTS: Only one of four goats in Group A had any bony ingrowth into the tantalum. Three of four goats in Group B demonstrated bony ingrowth. The average extent of bony ingrowth at the perimeter of the tantalum in Group A was 2.5% compared with 12.5% in Group B. Similarly, the volume of bony ingrowth within the tantalum was 2.5% in Group A and 10% in Group B. The difference was not statistically significant. CONCLUSIONS: The data in this pilot study suggest that tantalum may function as a synthetic osteoconductive bone graft substitute. The addition of rhBMP-2 may facilitate osteoinduction within a synthetic osteoconductive implant. The sample size in this study was too small for statistical significance. The present animal model as used in this study was inadequate for cervical arthrodesis where rigid implant fixation is desired.

Histomorphometric evaluation of the effects of ovariectomy on bone turnover in rat caudal vertebrae.

The purpose of this study was to learn whether caudal vertebrae can be used to evaluate the effects of ovariectomy (OVX) in rats. Seven-month-old female Wistar rats were divided into two groups: the OVX group and the untreated control group. All rats were killed at 8 weeks and their 4th lumbar (L4), 1st caudal (C1), 3rd caudal (C3), and 5th caudal (C5) vertebrae were processed undecalcified and sectioned with Villanueva bone stain for quantitative bone histomorphometry. Both length of vertebral bodies and the cancellous tissue area in C1 were similar in size to L4 but significantly bigger than C3 and C5. Within the groups, cancellous bone volume (BV/TV) and trabecular thickness in both groups gradually increased in caudal vertebrae in relation to the distal direction. Between the groups, OVX rats exhibited a significantly lower BV/TV relative to control rats at L4 and C1, however, no significant difference were seen at C3 and C5. Bone formation-related parameters such as osteoid and mineralizing surface, and eroded surface were higher in the OVX group than in the control group in caudal as well as in lumbar vertebrae. By quantitative analysis of bone marrow composition, yellow marrow volume in C3 and C5 was significantly higher than that in L4 and C1, in both groups. Our results suggest that C1 is similar to L4 in size, bone turnover, and bone marrow composition. However, further experiments are needed to evaluate the possibility that C1 vertebra could be used as an alternative site for histomorphometric evaluation of bone changes in OVX rats.

[Retinoic acid-induced hyperostosis in the murine spine, particularly in the cervical spine].

In order to investigate the role of retinoic acid in inducing spinal hyperostosis, the murine spine was radiologically and histologically examined after a 24-week peroral administration of synthetic retinoic acid (etretinate). In the cervical spine, osteophytes on the ventral rims of the vertebral bodies were more developed after etretinate administration than in controls. Histologically, chondrocytes after etretinate administration proliferated in the anterior longitudinal ligaments at the junction of the ventro-caudal rim and the epiphyseal region of the C2 and C3 vertebral bodies. Endochondral ossification was a characteristic feature and immature chondrocytes and mesenchymal cells were found in the region. After etretinate administration, the incidence of proliferative changes was 67.6% at the C2 ventro-caudal rim, and 79.4% and 70.6%, respectively in the C2 and C3 ventral epiphyseal regions. These percentages were highly significant compared with controls, but they were not dosage-dependent. These findings suggested that retinoic acid was involved in the differentiation and proliferation of chondrocytes in ligaments.