RECOVERY OF INTERVERTEBRAL DISC AFTER ENZYME TREATMENT

酶治疗后椎间盘的恢复

• 批准号: 2078813
• 负责人: Theodore Richard Oegema
• 金额: $ 20.63万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-04-01 至 1997-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2078813
• 关键词: aging; autoradiography; biomarker; collagen; disease /disorder model; dogs; enzyme therapy; extracellular matrix; gene expression; human tissue; immunofluorescence technique; in situ hybridization; intervertebral disk surgery; phenotype; proteoglycan; spinal disk injury; tissue /cell culture

DESCRIPTION: (Adapted From Investigator's Abstract) Disc degeneration which causes significant disability and loss of the quality of life, involves cellular, nutritional and mechanical components. The investigators hypothesize that cellular changes occur early and will test two hypotheses. The first is that there is a loss of normal matrix producing phenotype of cells in the nucleus pulposus and an increase of a fibrotic phenotype. This may be reversible as a result of changes in environment or irreversible. This phenotype may also be more sensitive to catabolic cytokines. The second hypothesis is that notochordal cells, which are critical to the development of nucleus pulposus, also play a role either directly by synthesis of matrix components or indirectly by influencing the other cells in the nucleus delaying early changes; and their loss accelerates degeneration.In order to test the first hypothesis, which considerable preliminary data supports, both matrix producing cells (MPC) and nonmatrix producing cells (NMPC) will be isolated from human disc and characterized from normal and degenerative discs. Sensitivity to IGF-1, TGF-B1, il-1B, and TNF-a will be examined.Co-culture experiments will be used to determine if interactions between the two cell populations occur. Because proteoglycans (PGs) represent a major extracellular matrix component, show changes that are highly correlated with aging and degeneration, and have turnover rapid enough to reflect short-term changes in the matrix, PGs and their associated link proteins will be used as indicator molecules. Presence of hyaluronic acid-binding regions, the cleavage of the protein core in chondroitin sulfate (CS) or keratan sulfate (KS) rich regions, and the presence and ratios of link proteins 1, 2, and 3 will be examined. Collagen types will also be determine. To test hypothesis two, notochordal cells will be isolated from nonchondrodystrophoid canine nucleus pulposus, a unique source of large amounts of these cells, and their characteristics determined with respect to matrix synthesis, turnover, and response to the growth factors and cytokines. Co-cultures with MPC and/or NMPC will determine if there are interactions between the different cell types. Key observations will be confirmed with human notochordal cells.

描述：（改编自研究者摘要）椎间盘退变 导致严重残疾和生活质量下降， 涉及细胞、营养和机械成分。这 研究人员假设细胞变化发生得很早，并将进行测试 两个假设。 首先是法向矩阵有损失 髓核细胞产生表型并增加 纤维化表型。 由于变化的结果，这可能是可逆的 环境或不可逆转。 这种表型也可能更敏感 分解代谢细胞因子。 第二个假设是脊索 对髓核发育至关重要的细胞 直接通过基质成分的合成发挥作用或 通过影响细胞核中的其他细胞间接延迟早期 变化；它们的损失会加速退化。为了测试 第一个假设，大量的初步数据支持，两者 基质产生细胞（MPC）和非基质产生细胞（NMPC）将 从人类椎间盘中分离出来，并从正常和 退变的椎间盘。 对 IGF-1、TGF-B1、il-1B 和 TNF-a 的敏感性将 进行检查。共培养实验将用于确定是否 两个细胞群之间发生相互作用。 因为蛋白多糖 (PG) 是主要的细胞外基质 成分，显示出与衰老高度相关的变化 退化，并且周转速度足够快以反映短期 基质、PG 及其相关连接蛋白的变化将 用作指示分子。 透明质酸结合的存在 区域，硫酸软骨素 (CS) 中蛋白质核心的裂解 或硫酸角质素（KS）丰富的区域，以及存在和比例 将检查连接蛋白 1、2 和 3。 胶原蛋白类型也会 确定。 为了检验假设二，脊索细胞将 从非软骨营养不良犬髓核中分离出来，一种独特的 大量这些细胞的来源及其特征 确定基质合成、周转和响应 生长因子和细胞因子。 与 MPC 和/或 NMPC 共培养将 确定不同细胞类型之间是否存在相互作用。 关键观察结果将通过人类脊索细胞得到证实。