Yudoh Kazuo, Shishido Kiyoshi, Murayama Hideki, Yano Mitsunobu, Matsubayashi Kenji, Takada Hiroya, Nakamura Hiroshi, Masuko Kayo, Kato Tomohiro, Nishioka Kusuki
St. Marianna University School of Medicine, Kawasaki, Japan.
Arthritis Rheum. 2007 Oct;56(10):3307-18. doi: 10.1002/art.22917.
Studies have shown the roles of oxidative stress in the pathogenesis of osteoarthritis (OA) and induction of chondrocyte senescence during OA progression. The aim of this study was to examine the potential of a strong free-radical scavenger, water-soluble fullerene (C60), as a protective agent against catabolic stress-induced degeneration of articular cartilage in OA, both in vitro and in vivo.
In the presence or absence of C60 (100 microM), human chondrocytes were incubated with interleukin-1beta (10 ng/ml) or H2O2 (100 microM), and chondrocyte activity was analyzed. An animal model of OA was produced in rabbits by resection of the medial meniscus and medial collateral ligament. Rabbits were divided into 5 subgroups: sham operation or treatment with C60 at 0.1 microM, 1 microM, 10 microM, or 40 microM. The left knee joint was injected intraarticularly with water-soluble C60 (2 ml), while, as a control, the right knee joint received 50% polyethylene glycol (2 ml), once weekly for 4 weeks or 8 weeks. Knee bone and cartilage tissue were prepared for histologic analysis. In addition, in the OA rabbit model, the effect of C60 (10 microM) on degeneration of articular cartilage was compared with that of sodium hyaluronate (HA) (5 mg/ml).
C60 (100 microM) inhibited the catabolic stress-induced production of matrix-degrading enzymes (matrix metalloproteinases 1, 3, and 13), down-regulation of matrix production, and apoptosis and premature senescence in human chondrocytes in vitro. In rabbits with OA, treatment with water-soluble C60 significantly reduced articular cartilage degeneration, whereas control knee joints showed progression of cartilage degeneration with time. This inhibitory effect was dose dependent, and was superior to that of HA. Combined treatment with C60 and HA yielded a significant reduction in cartilage degeneration compared with either treatment alone.
The results indicate that C60 fullerene is a potential therapeutic agent for the protection of articular cartilage against progression of OA.
研究表明氧化应激在骨关节炎（OA）发病机制中发挥作用，并在OA进展过程中诱导软骨细胞衰老。本研究旨在探讨一种强效自由基清除剂——水溶性富勒烯（C60），在体外和体内作为一种保护剂，抵抗分解代谢应激诱导的OA关节软骨退变的潜力。
在有或没有C60（100微摩尔）存在的情况下，将人软骨细胞与白细胞介素-1β（10纳克/毫升）或过氧化氢（100微摩尔）一起孵育，并分析软骨细胞活性。通过切除内侧半月板和内侧副韧带在兔中建立OA动物模型。将兔分为5个亚组：假手术组或用0.1微摩尔、1微摩尔、10微摩尔或40微摩尔的C60治疗组。左膝关节关节腔内注射水溶性C60（2毫升），而作为对照，右膝关节接受50%聚乙二醇（2毫升），每周一次，共4周或8周。制备膝关节骨和软骨组织用于组织学分析。此外，在OA兔模型中，将C60（10微摩尔）对关节软骨退变的影响与透明质酸钠（HA）（5毫克/毫升）的影响进行比较。
C60（100微摩尔）在体外抑制了分解代谢应激诱导的基质降解酶（基质金属蛋白酶1、3和13）的产生、基质产生的下调以及人软骨细胞的凋亡和早衰。在患有OA的兔中，用水溶性C60治疗显著减少了关节软骨退变，而对照膝关节显示软骨退变随时间进展。这种抑制作用是剂量依赖性的，并且优于HA。与单独任何一种治疗相比，C60和HA联合治疗使软骨退变显著减少。
结果表明C60富勒烯是一种潜在的治疗剂，可保护关节软骨免受OA进展的影响。
