SUPEROXIDE AND BONE RESORPTION

这项建议的广泛的长期目标是了解 超氧化物在破骨细胞介导的骨吸收中的作用。 我们的关系 在皱褶边缘间隙内骨细胞超氧化物生成之间 和骨吸收; 2.以确定超氧化物的影响 产生于骨细胞内的超氧化物 细胞膜或外部或两者; 3.为了确定细胞因子， 干扰素γ，已知也刺激白细胞中的超氧化物生成 从慢性肉芽肿病患者，增强破骨细胞- 介导的骨吸收和骨硬化小鼠突变体，如果是这样， 定位作用部位。 在这些实验中， 将在分离的破骨细胞培养物中监测超氧化物清除剂 和头盖骨 比较从细胞中排除的清道夫 将膜按尺寸转化为低分子量清除剂， 膜，超氧化物对骨吸收的影响将是 寻求。 通过使用不同年龄的新生动物，我们将评估 破骨细胞形成速率的重要性 氧自由基影响骨吸收。 NBT染色将用于 测量和定位个体中氧自由基产生的变化 破骨细胞 干扰素γ将用于刺激超氧化物 骨硬化混合症中破骨细胞的生成。 如果干扰素γ 刺激破骨细胞介导的骨吸收， 骨硬化动物，超氧化物清除剂将被添加到相同的 培养，以确定超氧化物生成的贡献 增加骨吸收能力。 这些实验已经 为干扰素γ治疗的I/II期试验提供了基础 骨硬化患者。

The broad long-tern objective of this proposal is to understand the role of superoxide in osteoclast-mediated bone resorption. Our relationship between osteoclastic superoxide generation within the ruffled border space and bone resorption; 2. to determine if the effect(s) of superoxide produced in bone is due to superoxide generated within the osteoclastic cellular membrane or outside or both; 3. to determine if a cytokine, Interferon Gamma, known too stimulate superoxide generation in leukocytes from patients with chromic granulomatous disease, enhances osteoclast- mediated bone resorption in and osteopetrotic mouse mutant and if so, to localize the site of action. In these experiments, the effects of superoxide scavengers will be monitored in cultures of isolated osteoclasts and calvaria. Comparing a scavenger which is excluded from cellular membranes by size to a low molecular weight scavenger which permeates membranes, the site of superoxide's effects upon bone resorption will be sought. By using neonatal animals of different ages, we shall assess the importance of the rate of osteoclasts formation upon the mechanism by which oxygen radicals affect bone resorption. NBT staining will be used to measure and localize changes in oxygen radical production in individual osteoclasts. Interferon Gamma will be used to stimulate superoxide generation in osteoclasts from osteopetrotic misc. If Interferon Gamma stimulates osteoclast-mediated bone resorption in tissues cultured from osteopetrotic animals, superoxide scavengers will be added to identical cultures to determine the contribution of superoxide generation to the increased bone resorptive capacity. These experiments have already provides the basis for a phase I/II trial of Interferon Gamma to treat osteopetrotic patients.