• Journal of Korean Neurosurgical Society
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• v.48 no.1
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• pp.1-7
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• 2010

Objective : Notochordal cells in the intervertebral disc interact with nucleus pulposus (NP) cells and support the maintenance of disc homeostasis by regulation of matrix production. However, the influence of notochordal cells has not been evaluated in the annulus fibrosus (AF), which is the primary pain generator in the disc. We hypothesized that the notochordal cell has the capacity to modulate inflammatory mediators secreted by AF cells secondary to stimulation. Methods : Notochordal and AF cells were isolated from adult New Zealand white rabbits. AF pellets were cultured with notochordal cell clusters or in notochordal cell-conditioned media (NCCM) for 24 or 48 hours with proinflammatory cytokines at varying concentrations. Gene expression in AF pellets were assayed for nitric oxide synthase (iNOS), cyclo-oxygenase (COX)-2, and interleukin (IL)-6 by real time reverse transcriptase polymerase chain reaction (RT-PCR). Results : AF pellet in NCCM significantly decreased the iNOS and COX-2 messenger ribonucleic acid (mRNA) levels compared to AF pellets alone and AF pellets with notochordal cells (p < 0.05). AF pellet resulted in dose-dependent iNOS and COX-2 expression in response to IL-$1{\beta}$, stimulation, demonstrating that 1 ng/ml for 24 hours yielded a maximal response. AF pellet in NCCM significantly decreased the expression of iNOS and COX-2 in response to 1ng/ml IL-$1{\beta}$, stimulation at 24 hours (p < 0.05). There was no difference in IL-6 expression compared to AF pellets alone or AF pellets with notochordal cell clusters. Conclusion : We conclude that soluble factors from notochordal cells mitigate the gene expression of inflammatory mediators in stimulated AF, as expected after annular injury, suggesting that notochordal cells could serve as a novel therapeutic approach in symptomatic disc development.

• Applied Microscopy
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• v.21 no.2
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• pp.39-56
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• 1991

The development of notochordal cells of nucleus pulposus was studied with electron microscope in human fetuses ranging from 30 mm to 260 mm crown-rump length. At 30 mm fetus, primitive notochordal cells were large with central nucleus, few organelles, and their cytoplasm usually contained dense glycogen and fine filaments. Notochordal cells at all ages contained bundles of fine filaments of indeterminate nature. One unusual feature of fetal notochordal cells was the consistent presense of rough endoplasmic reticulum surrounding poorly developed mitochondria. At 50 mm fetus, notochordal cells formed dense masses with interdigitating cell membranes connected by a variety of cell to cell junctions. With increasing age, the cell connections became slender threaded cytoplamic extending from cell and enclosed large extracellular space. Chondrocyte-like cells appeared to be separated by large volumes of extracellular matrix. Viable notochordal and condrocyte-like cells existed in specimen from all age. The extracellular spaces were filled with fibrillar and granular material by 90 mm fetus. Necrotic cells were distinguished by loss of their membrane integrity, vacuolization of their organelles, and the presence of dense osmiophilic masses. In adult tissue, notochordal cells became rounded or irregular in shape and developed a pericellular matrix consisting of collagen fibrile, and dense particle. The structure of notochordal cells and their persistance in the nucleus pulposus after fetal life suggested that they may have a significant role in the formation and maintenance of the nucleus pulposus. The presence of Golgi complex and well-developed endoplasmic reticulum in chondrocyte-like cells suggested that they are capable of producing and maintaining the extracellular matrix.

• Journal of Korean Neurosurgical Society
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• v.55 no.5
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• pp.237-243
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• 2014

Objective : Symptomatic disc degeneration develops from inflammatory reactions in the annulus fibrosus (AF). Although inflammatory mediators during annular inflammation have been studied, the roles of matrix metalloproteinases (MMPs) and their inhibitors have not been fully elucidated. In this study, we evaluated the production of MMPs and tissue inhibitors of metalloproteinase (TIMPs) during annular inflammation using an in vitro co-culture system. We also examined the effect of notochordal cells on annular inflammation. Methods : Human AF (hAF) pellet was co-cultured for 48 hours with phorbol myristate acetate-stimulated macrophage-like THP-1 cells. hAF pellet and conditioned media (CM) from co-cultured cells were assayed for MMPs, TIMPs, and insulin-like growth factor (IGF)-1 levels using real-time reverse-transcriptase polymerase chain reaction and enzyem-linked immunosorbent assay. To evaluate whether notochordal cells affected MMPs or TIMPs production on annular inflammation, hAF co-cultured with notochordal cells from adult New Zealand White rabbits, were assayed. Results : MMP-1, -3, -9; and TIMP-1 levels were significantly increased in CM of hAF co-cultured with macrophage-like cells compared with hAF alone, whereas TIMP-2 and IGF-1 levels were significantly decreased (p<0.05). After macrophage exposure, hAF produced significantly more MMP-1 and -3 and less TIMP-1 and -2. Interleukin-$1{\beta}$ stimulation enhanced MMP-1 and -3 levels, and significantly diminished TIMP-2 levels. Co-culturing with rabbit notochordal cells did not significantly influence MMPs and TIMPs production or COL1A2 gene expression. Conclusion : Our results indicate that macrophage-like cells evoke annular degeneration through the regulation of major degradative enzymes and their inhibitors, produced by hAF, suggesting that the selective regulation of these enzymes provides future targets for symptomatic disc degeneration therapy.

• The Korean Journal of Cytopathology
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• v.6 no.2
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• pp.199-203
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• 1995

Chordoma is relatively uncommon tumor comprising $1\sim4%$ of primary malignant bone tumors, and believed to arise from the remnants of notochordal tissue. Because of its rare occurrence in the thoracic spine, we report a case of chordoma involving the thoracic spine. A 45-year-old male was sufferred from chest pain radiating to the back. Chest CT showed a well marginated, round huge mass with multiseptated enhancement at the thoracic spine from T5 to T8 level. After percutaneous needle aspiration, piecemeal resection of the tumor was done. On cytologic smears, two types of neoplastic cells were arranged in sheets and cords in mucinous background. One type of cells consisted of medium sized cells with pink cytoplasm and round nuclei. The other type had voluminous bubbly or clear cytoplasm divided by intracytoplasmic septae imparting a feathery or basket-like appearance. Histologically, the tumor showed lobulated feature divided by fibrous septae and the tumor cells were pink eosinophilic or physaliphorous in morphology. Immunohistochemically, the tumor cells revealed strong positivity for low(AE1) and high (AE3) molecular weight cytokeratins.

• The Korean Journal of Cytopathology
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• v.8 no.2
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• pp.199-203
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• 1997

Chordoma is an uncommon neoplasm that accounts for approximately 1% to 4% of all primary bone neoplasms and thought to originate from remnants of the fetal notochordal elements. It usually occurs in adults and has a predilection for the sacrococcygeal and spheno-occipital areas. Chondroid chordoma, first described by Heffelfinger et al, is a rare variant of chordoma; it contains both chordomatous and chondromatous features, and has a considerably better prognosis than either chordoma or chondrosarcoma. The cytologic findings of fine needle aspiration of sacrococcygeal chondroid chordoma in a 57-year-old man are presented. Aspiration cytology showed many sheets and cords of neoplastic cells in a thick amorphous blue-purple mucinous background. The cells had small too medium sized round nuclei with coarse granular chromatin and abundant eosinophilic or bubbly cytoplasm. Some cells had pleomorphic and hyperchomatic nuclei with prominent nucleoli. Cytologic findings were compared to histologic findings. Histologically, areas of chondroid differentiation were noted which were absent in the cytologic smear. Immunohistochemically, both the chondroid and chordoid areas had an epithelial phenotype and stained for cytokeratin, epithelial membrane antigen and S-100 protein. This is the first case of cytologic findings of chondroid chordoma to our knowledge in literature.

• Korean Journal of Veterinary Research
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• v.37 no.1
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• pp.59-69
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• 1997

The histological and histochemical study of the body of axis and 1st intervertebral disc in the Korean native cattle fetus was attempted to early developmental process. The experimental animals used in this study were collected from fetus of the Korean native cattle ranging from 50mm to 180mm in Crown-Rump length(CR-length). The results were summerized as follows: 1. The ossification centers appeared centrum 1 and centrum 2 of the axis in 110mm CR-length fetus of the Korean native cattle. The centrum 2 was formed little earlier than the centrum 1. 2. The histochemical reactions for collagenous fibers in the axis revealed negative in 50mm CR-length, mild positive in 90mm CR-length, and strong positive in 110mm CR-length, respectively. 3. Dense collagenous fibers were observed in the notochord through the centrum 2, and intervertebral disc developed into cuneiform dorsoventrally, It's contour looks like an annual ring. These fiber bundle arranged lamellar formation. 4. The intervertabral disc of 50mm CR-length fetus was composed mainly mesenchymal cells, and these cell showed aggregation in the central portion. The intervertebral disc of 110mm CR-length fetus was consisted of pricipally fibroblast, and notochordal sheath formed with two layers in the center.