Extracellular matrix in bone and cartilage destruction.

骨和软骨中的细胞外基质被破坏。

• 批准号: 08407048
• 负责人: IWATA Hisashi
• 金额: $ 19.9万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08407048/
• 关键词: matrix metalloproteinases; tissue inhibitors of metalloproteinase; chemokines; bone cement interface tissue; type I procollagen; 1,25-dihydroxyvitamin D; posttranslational control; chemonucleolysis; 骨形成

Matrix metalloproteinases (MMPS) and tissue inhibitors of metalloproteinase (TIMPs) play an important role in tissue destruction and remodeling. We focused the synvial fluid and interface tissue of loosening THA.We revealed. these points.Extremely high concentrations of MMP-3 in synovial fluid (SF) of the patients with RA may contribute to its elevation in serum. It would seem that regulation of TIMP production depends upon the dis-ease state and not the concentration of MMP.Discrepancies between concentrations of TIMP and MMP in SF may be responsible for cartilage destruction in RA.The samples of cement inter-face tissues from patients who had failed cemented total hip arthroplasty (THA) were obtained for revision of THA and analyzed on mRNA expression of MMPs and TIMPs. We used the revers transcrip-tional polymerase chain reaction (RT-PCR). mRNA of MMP--1, -2, -3, -9, and TIMP-1 and -2 was detected in the interface tissue. MMP-10 mRNA was not detected, yet MMP-1 and MMP-3 mRNA were c … More ommonly observed.TIMT-2 mRNA was also strongly expressed compared to TIMP-1. It was thus demonstrated that MMPs and TIMPs were produced locally in the cement bone interface tissue of THA loosening.Also we revealed that polyethylene debris sur-rounded by macrophages and phagocytosis of debris by macrophages was frequently observed in the interface tissue. Macrophage activation and the production of inflammatory cytokines such as IL-1 and TNFa might induce the development of interface tissue. Expression of chemokine mRNAs was also commonly seen, suggesting that this led to recruitment of macrophages into the bone cement interface tissue. Debris released from implants appears to cause activation of macrophages and the production of inflammatory cytokines and chemokines that induce cellular recruitment into interface tissue. We suggested the mechanism might form a vicious cycle that aggravates THA loosening.We tryed to develop new treatment of disc herniation using chondroitinase ABC.Experimental chemonucleolysis with chondroitinase ABC as compared with chymopapain, was investigated in monkeys. The effects of these two enzyms were analyzed morphologically and biochemically. The results confirm that selective degradation is achived with chondroitinase ABC in monkeys and that chondroitinase ABC is less toxic to discs than chymopapain is.Also, we investigated the posttranscriptional regulation of tissue matrix. So, the kinetics of type I procollagen synthesis in a human osteosarcoma cell line, MG 63, were investigated after treatment with 1,25-dihydroxyvitamin D.The results therefore suggest an increase in both the synthesis and secretion of type I collagen. Moreover, gelatinase B-resistant procollagen molecules, indicative of intracellular procollagen molecules in the stable triple helical form, were detected only in the 1,25-(OH)2 D,-treated cells. The present evidence points to posttranslational control of procollagen synthesis. Less

基质金属蛋白酶(MMPs)及其组织抑制因子(TIMPs)在组织破坏和重塑中发挥着重要作用。我们将松动THA的突触液和界面组织集中在一起。这些观点：RA患者滑液中极高浓度的基质金属蛋白酶-3可能与其在血清中的升高有关。TIMP产生的调节似乎依赖于疾病状态，而不是MMPs的浓度。TIMP和MMPs在SF中的浓度差异可能是RA软骨破坏的原因。采用逆转录-聚合酶链式反应(RT-PCR)。在界面组织中检测到基质金属蛋白酶-1、-2、-3、-9、TIMP-1和-2mRNA的表达。未检测到MMP10mRNA，而MMP1和MMP3mRNA均为c…与TIMP-1相比，TIMT-2mRNA的表达也更强。结果表明，松动时骨水泥界面组织局部产生MMPs和TIMP，界面组织中可见巨噬细胞包裹的聚乙烯碎屑和巨噬细胞吞噬碎屑的现象。巨噬细胞的激活和IL-1、TNFa等炎性细胞因子的产生可能诱导界面组织的发育。趋化因子mRNAs的表达也很常见，这表明这导致巨噬细胞重新聚集到骨水泥界面组织中。植入物释放的碎片似乎会激活巨噬细胞，产生炎性细胞因子和趋化因子，导致细胞重新聚集到界面组织中。我们认为这一机制可能形成了一个恶性循环，加剧了松动。我们试图开发使用软骨素酶ABC治疗腰椎间盘突出症的新方法。对这两种酶的作用进行了形态和生化分析。结果证实了软骨素酶ABC在猴子体内实现了选择性降解，并且软骨素酶ABC对椎间盘的毒性比木瓜凝乳酶要小。此外，我们还研究了组织基质的转录后调节。因此，我们研究了人骨肉瘤细胞系MG 63经1，25-二羟基维生素D处理后I型前胶原合成的动力学，结果提示I型胶原的合成和分泌均增加。此外，明胶酶B抗性的前胶原分子，表明细胞内的前胶原分子以稳定的三螺旋形式存在，仅在1，25-(OH)2D处理的细胞中检测到。目前的证据表明，翻译后控制了前胶原的合成。较少

项目成果

Toshiki Iwase, Y.Hasegawa, T.Ito, N.Makihara, H.Takahashi, Hisashi Iwata: "Bone composition and metabolism after hyperbaric oxygenation in rats with 1-hydroxyethylidene-1,1-bisphosphonate-induced rickets" Undersea Hyperb Med. 23. 5-9 (1996)

Toshiki Iwase、Y.Hasekawa、T.Ito、N.Makihara、H.Takahashi、Hisashi Iwata：“1-羟基亚乙基-1,1-二磷酸盐诱发佝偻病大鼠高压氧合后的骨成分和代谢”Undersea Hyperb Med。

T.Kajima, K.Kozaki, S.Saga, Y.Hashizume, Naoki Ishiguro, Hisashi Iwata, O.Miyaishi: "Alteration of the kinetics of Type I procollagen synthesis in human osteosarcoma cells by 1,25-Dihydroxyvitamin D3" J Cell Biochem. 65. 542-549 (1997)

T.Kajima、K.Kozaki、S.Saga、Y.Hashizume、Naoki Ishiguro、Hisashi Iwata、O.Miyaishi：“1,25-二羟基维生素 D3 改变人骨肉瘤细胞中 I 型前胶原合成的动力学” J Cell

K.Kurita,T.Shinomura,M.Ujita,M.Zako,D.Kida,H.Iwata,K.Kimata: "Occurrence of PG-Lb,a leucine-rich small chondroitin/dermatan sulphate,proteoglycan in mammaliam epiphyseal cartilage : molecular cloning and sequence analysis of the mouse cDNA" Biochem.J.318.

K.Kurita、T.Shinomura、M.Ujita、M.Zako、D.Kida、H.Iwata、K.Kimata：“哺乳动物骨骺软骨中富含亮氨酸的小软骨素/硫酸皮肤素、蛋白多糖 PG-Lb 的出现

Yasushi Miura, K.Mimatsu, Hisashi Iwata: "Massive tongue swelling as a complication after spinal surgery" J Spinal Disord. 9. 339-341 (1996)

Yasushi Miura、K.Mimatsu、Hisashi Iwata：“脊柱手术后的并发症是舌头大面积肿胀”J Spinal Disord。

Toshiki Iwase, Y.Hasegawa, T.Ito, N.Makihava, H.Takahashi, Hisashi Iwata: "Bone composition and metabolism after hyperbaric oxygenation in rats with 1-hydroxyethylidene-1,1-bisphosphonate-induced rickets" Undersea Hyperb Med. 23. 5-9 (1996)

Toshiki Iwase、Y.Hasekawa、T.Ito、N.Makihava、H.Takahashi、Hisashi Iwata：“1-羟基亚乙基-1,1-二磷酸盐诱发佝偻病大鼠高压氧合后的骨成分和代谢”Undersea Hyperb Med。