Prevention of joint contracture following joint immobilization using gene therapy's method

利用基因疗法预防关节固定后的关节挛缩

基本信息

批准号：
13671507
负责人：
HASHIMOTO Jun
金额：
$ 2.3万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2002
项目状态：
已结题

项目摘要

Joint contracture followed by prolonged joint immobilization is a serious problem in orthopaedic treatment. The cause of that was reported as that proliferation of fibrous synovial tissue in the joint cavity was characteristic. To prevent the contracture followed by immobilization, we planned to suppress the proliferation of synoviocytes by gene transfer of transcription factor E2F decoy into synoviocytes. As the animal model of joint immobilization, we used Wilson's rat model (Wilson et al Clin Orthop 1988). At 1, 2, 3 and 4 weeks after joint immobilization, we examined the Hematoxylin and eosin staining of the sagittal section of knee joint and confirmed the proliferation of synovial tissue and adhesion of extracellular matrix in the joint reproductively. In vitro study, we examined the semi-quantitative transfection efficiency of gene transfer into fibroblast-like cell line (3Y1-B and NRK) and primary generated rat synoviocyte using FITC-labeled oligonucleotide (FITC ODN) included in HVJ-envelope vector, and determined the optimal condition of gene transfer. In vivo, we observed the high efficient gene transfer into synovial tissue by intraarficular injection of FITC ODM included in HVJ envelope vector, and almost no transfer into other tissue such as articular cartilage, meniscus and ligament. Now we examine in vivo gene transfer using E2F decoy included in HVJ envelope and analyze the proliferation of synovial tissue and production of extracellular matrix by molecular biologically and histologically.

关节缔合，然后延长关节固定是骨科治疗中的一个严重问题。据报道，关节腔中纤维滑膜组织的增殖是特征的。为了防止缔合之后固定，我们计划通过转录因子E2F诱饵转移到滑膜细胞中来抑制滑膜细胞的增殖。作为关节固定的动物模型，我们使用了威尔逊的大鼠模型（Wilson等Clin Orthop 1988）。关节固定后的1、2、3和4周，我们检查了膝关节矢状片的苏木精和曙红染色，并证实了滑膜组织的增殖以及关节中细胞外基质的粘附。在体外研究中，我们使用FITC标记的寡核苷酸（FITC ODN）检查了基因转染到成纤维细胞样细胞系（3Y1-B和NRK）中的半定量转染效率，并确定了基因转移的最佳条件。在体内，我们通过缺乏HVJ包膜载体中的FITC ODM的缺乏症状注射来观察到高效的基因转移到滑膜组织中，几乎没有转移到其他组织中，例如关节软骨，半月板和韧带。现在，我们使用HVJ包膜中的E2F诱饵检查体内基因转移，并通过分子生物学和组织学上分析滑膜组织的增殖和细胞外基质的产生。