BIOCHEMISTRY OF INTERVERTEBRAL DISC

椎间盘的生物化学

• 批准号: 3157729
• 负责人: David R Eyre
• 金额: $ 10.12万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-01-01 至 1988-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3157729
• 关键词: aging; biochemistry; crosslink; high performance liquid chromatography; histopathology; human tissue; intervertebral disk; molecular pathology; pain; spinal cord disorders

Most experts are convinced that degenerating intervertebral discs are at the root of many cases of low-back pain. The intervertebral disc has only just begun to be studied in detail at the molecular level and is still a poorly characterized connective tissue. We propose to characterize in more detail the collagen of human and animal intervertebral discs using the latest biochemical techniques. The emphasis will be an attempt to relate structure to function. An important aim is to understand the characteristic changes in human disc properties with age. The work on human discs will focus initially on autopsy specimens of lumbar discs carefully selected on the basis of histopathological examination to be typically normal for their age, with no gross lesions. Rabbit and cow lumbar discs will also be studied. Molecular species of collagen will be mapped quantitatively within the annulus fibrosus and nucleus pulposus, including the bulk types I and II collagens that are known to be present and the AlphaA, AlphaB, 1Alpha, 2Alpha, and 3Alpha species which may be present. Site-related changes in the distribution of these collagen species will be correlated with variations in the distribution and extractability of proteoglycans and non-collagenous proteins, and with swelling properties of the tissue. Crosslinking residues of collagen will be isolated and quantified, including hydroxypyridinium residues, the mature forms. Pepsin-solubilized collagen will be fractionated into individual species, type I, type II, etc., and the level and type of crosslink in each genetic species and other chemical properties will be measured. Using high performance liquid chromatography (HPLC) to fractionate collagen peptides, any alterations in the distribution of hydroxypyridinium crosslinks with age and location in the disc will be assessed. The molecular packing of annulus and nucleus collagen will be examined by low angle X-ray diffraction and electron microscopy seeking, for example, age-related changes in hydration. Discs removed at surgery to correct scoliosis and other conditions, including spondylolisthesis are on hand and will continue to be available and will also be studied. A collaboration will be initiated with the biomechanics laboratory at Beth Israel Hospital to relate the visco-elastic shear modulus of disc specimens with their composition. In summary, the objective is a better understanding of the relationship between molecular structure, age-related degeneration and mechanical failure.

大多数专家相信，退变的椎间盘是在 许多腰痛的根源。 椎间盘只有 刚刚开始在分子水平上进行详细研究，并且仍然是一个 结缔组织特征较差。 我们建议更多地描述 使用以下方法详细介绍人类和动物椎间盘的胶原蛋白 最新的生化技术。 重点是尝试联系 结构到功能。 一个重要的目标是了解 人类椎间盘特性随年龄的变化。 工作内容 人类椎间盘将首先关注腰椎间盘的尸检标本 在组织病理学检查的基础上精心挑选 对于他们的年龄来说通常是正常的，没有明显的病变。 兔子和牛 腰椎间盘也将被研究。 胶原蛋白的分子种类将是 定量绘制纤维环和髓核内的图， 包括已知存在的大量 I 型和 II 型胶原蛋白 以及 AlphaA、AlphaB、1Alpha、2Alpha 和 3Alpha 物种，它们可能是 展示。 这些胶原蛋白分布的位点相关变化 物种将与分布的变化相关 蛋白聚糖和非胶原蛋白的可提取性，以及 组织的膨胀特性。 胶原蛋白的交联残留物将 被分离和定量，包括羟基吡啶鎓残留物， 成熟的形式。 胃蛋白酶溶解的胶原蛋白将被分离成 个别物种、I 型、II 型等，以及水平和类型 每个遗传物种中的交联和其他化学特性将 测量。 使用高效液相色谱 (HPLC) 分级胶原蛋白肽，分布的任何改变 羟基吡啶鎓会随着年龄和椎间盘位置而交联 评估。 纤维环和纤维核胶原蛋白的分子堆积将是 通过低角度 X 射线衍射和电子显微镜寻找进行检查， 例如，与年龄相关的水合作用变化。 手术中取出椎间盘 矫正脊柱侧弯和其他病症，包括脊椎滑脱 手并将继续可用并且也将被研究。 一个 将与贝斯生物力学实验室启动合作 以色列医院将椎间盘标本的粘弹性剪切模量联系起来 与他们的组成。 总而言之，目标是更好 了解分子结构之间的关系，与年龄相关 退化和机械故障。