Influence of tissue maturation and antioxidants on the apoptotic response of articular cartilage after injurious compression
Influence of tissue maturation and antioxidants on the apoptotic response of articular cartilage after injurious compression
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DOI：
10.1002/art.11438
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发表时间：
2004-01-01
影响因子：
--
通讯作者：
Schünke, M
中科院分区：
文献类型：
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作者：
Kurz, B;Lemke, A;Schünke, M
目标。目的：研究组织成熟度和抗氧化剂对损伤压迫诱导的牛关节软骨细胞凋亡的影响。取0.5~23月龄牛关节软骨盘，取材于股骨距骨沟内培养。将软骨圆盘与模拟细胞通透性的超氧化物歧化酶(SOD)模拟锰(0~12.5微米)或α-生育酚(0~50微米)预孵育过夜，然后以1 mm/s的速度对软骨盘进行一次无约束压缩，最终应变为50%。附加孵育4天后，固定并包埋磁盘进行光镜和电子显微镜检查。通过光镜下核泡的出现对凋亡细胞进行形态定量。通过掺入放射性标记的脯氨酸来证明其生物合成活性。通过与四氮基蓝孵育后的软骨组织学检查，证实了超氧化物歧化酶模拟物的抗氧化作用。损伤压迫诱导新生小牛软骨盘的细胞凋亡率(22%)显著高于成熟奶牛软骨盘的凋亡率(2-6%)。在22月龄动物的软骨中，超氧化物歧化酶模拟物以剂量依赖的方式降低损伤诱导的细胞凋亡率(2.5微米完全抑制)，而α-生育酚无作用。抗氧化剂不会改变蛋白质的生物合成或细胞超微结构。我们的数据表明，关节软骨对机械损伤的凋亡反应受成熟度的影响，部分是由活性氧介导的。组织的抗氧化状态可能是防止关节软骨机械性细胞死亡的重要因素。
Objective. To study the influence of tissue maturation and antioxidants on apoptosis in bovine articular cartilage induced by injurious compression.Methods. Bovine articular cartilage disks were obtained from the femoropatellar groove of animals ages 0.5-23 months and placed in culture. Cartilage disks were preincubated overnight with the cell-permeable superoxide dismutase (SOD) mimetic Mn(III) porphyrin (0-12.5 muM) or alpha-tocopherol (0-50 muM) and then injured by a single unconfined compression to a final strain of 50% at a velocity of 1 mm/second. After 4 days of additional incubation, the disks were fixed and embedded for light and electron microscopy. Apoptotic cells were quantified morphologically by the appearance of nuclear blebbing on light microscopy. Biosynthetic activity was demonstrated by incorporation of radiolabeled proline. The antioxidative action of the SOD mimetic was confirmed by histologic examination of cartilage after incubation with nitroblue tetrazolium.Results. Injurious compression induced significantly more apoptosis in cartilage disks from newborn calves (22% of cells) than in cartilage from more mature cows (2-6%). In cartilage from 22-month-old animals, the SOD mimetic reduced the percentage of apoptotic cells induced by injury in a dose-dependent manner (complete inhibition with 2.5 muM), while alpha-tocopherol had no effect. Neither antioxidant altered protein biosynthesis or cellular ultrastructure.Conclusion. Our data suggest that the apoptotic response of articular cartilage to mechanical injury is affected by maturation and is mediated in part by reactive oxygen species. The antioxidative status of the tissue might be important for the prevention of mechanically induced cell death in articular cartilage.