Nucleus pulposus notochord cells secrete connective tissue growth factor and up-regulate proteoglycan expression by intervertebral disc chondrocytes
Nucleus pulposus notochord cells secrete connective tissue growth factor and up-regulate proteoglycan expression by intervertebral disc chondrocytes
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DOI：
10.1002/art.22258
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发表时间：
2006-12-01
影响因子：
--
通讯作者：
Inman, Robert D.
中科院分区：
文献类型：
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作者：
Erwin, W. Mark;Ashman, Keith;Inman, Robert D.
Objective.鉴定从椎间盘髓核衍生的犬脊索细胞获得的条件培养基的组分，并评估这些因素影响椎间盘衍生的软骨细胞聚集蛋白聚糖、多功能蛋白聚糖和透明质酸合成酶2（HAS-2）基因表达的能力，作为培养条件的函数。方法.在血清缺乏条件下（Dul-becco改良Eagle培养基[DMEM]），在藻酸盐珠中培养从非软骨营养不良犬获得的犬脊索细胞，以制备脊索细胞条件培养基（NCCM）。采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和液相色谱-串联质谱法对NCCM进行评价。用无血清培养基（DMEM）和NCCM培养牛椎间盘来源的软骨细胞，并测定组织培养条件对聚集蛋白聚糖、多功能蛋白聚糖和HAS-2基因表达的影响。接下来，使用含有重组结缔组织生长因子（rCTGF）的DMEM评估软骨细胞的聚集蛋白聚糖基因表达，并将结果与使用NCCM和DMEM获得的结果进行比较。结果NCCM含有聚集蛋白聚糖、Cu/Zn超氧化物歧化酶、纤连蛋白和CTGF前体。NCCM培养引起聚集蛋白聚糖、多功能蛋白聚糖和HAS-2基因表达的上调。NCCM诱导软骨细胞中聚集蛋白聚糖基因表达的水平与100-200 ng/ml rCTGF诱导的水平相似。非软骨营养不良和软骨营养不良犬脊索细胞表现出相似的CTGF基因表达水平。结论髓核衍生的脊索细胞分泌CTGF（CCN 2），其是最近发现的多功能生长因子。CTGF基因在非软骨营养不良和软骨营养不良犬脊索细胞中的表达没有差异，这表明CTGF作为椎间盘核内的合成代谢因子的可能作用，至少在一定程度上依赖于椎间盘核内的脊索细胞群。
Objective. To identify the components of conditioned medium obtained from intervertebral disc nucleus pulposus-derived canine notochord cells, and to evaluate the capacity of such factors to affect disc-derived chondrocyte gene expression of aggrecan, versican, and hyaluronic acid synthase 2 (HAS-2) as a function of culture conditions. Methods. Canine notochord cells obtained from nonchondrodystrophic dogs were cultured within alginate beads under conditions of serum deficiency (Dul-becco's modified Eagle's medium [DMEM]) to produce notochord cell-conditioned medium (NCCM). NCCM was evaluated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and liquid chromatography-tandem mass spectroscopy. Bovine disc-derived chondrocytes were cultured with serumdeficient medium (DMEM) and NCCM and assayed for the effect of tissue culture conditions on aggrecan, versican, and HAS-2 gene expression. Next, chondrocyte gene expression for aggrecan was evaluated using DMEM containing recombinant connective tissue growth factor (rCTGF), and the results compared with those obtained using NCCM and DMEM. Results. NCCM contained aggrecan, Cu/Zn superoxide dismutase, fibronectin, and CTGF precursor. Culture with NCCM caused an up-regulation of aggrecan, versican, and HAS-2 gene expression. NCCM induced aggrecan gene expression in chondrocytes at a level similar to that induced by 100-200 ng/ml rCTGF. Nonchondrodystrophic and chondrodystrophic canine notochord cells exhibited similar levels of CTGF gene expression. Conclusion. Nucleus pulposus-derived notochord cells secrete CTGF (CCN2), a recently discovered multifunctional growth factor. There is no difference between CTGF gene expression in nonchondrodystrophic and chondrodystrophic canine notochord cells, suggesting a possible role of CTGF as an anabolic factor within the disc nucleus that is, to at least some degree, dependent on the population of notochord cells within the disc nucleus.