In vivo transfer of interleukin-1 receptor antagonist gene in osteoarthritic rabbit knee joints -: Prevention of osteoarthritis progression
In vivo transfer of interleukin-1 receptor antagonist gene in osteoarthritic rabbit knee joints -: Prevention of osteoarthritis progression
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DOI：
10.1016/s0002-9440(10)65368-0
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发表时间：
1999-04-01
影响因子：
6
通讯作者：
Pelletier, JP
中科院分区：
文献类型：
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作者：
Fernandes, J;Tardif, G;Pelletier, JP
本研究的目的是通过关节内质粒注射确定局部IL-1 Ra基因治疗对骨关节炎模型中骨关节切除术兔的结构变化的功效。通过内侧髌旁切口对家兔进行右膝关节部分切除术。然后将兔子分为四个实验组。第1组未接受治疗。第2组以24小时间隔连续三次接受含有脂质γ AP-DL-RIE/DOPE和DNA质粒VR 1012的0.9%盐水的关节内注射。第3组连续三次注射含有1000 μ g犬IL-1 Ra质粒和脂质的盐水。手术后4周开始注射。第1组于术后4周处死，其余各组于术后8周处死，分别对内侧髁、股骨髁、胫骨平台和滑膜的软骨组织进行大体和显微镜下观察。还使用抗犬IL-1 Ra的兔多克隆抗体对标本进行免疫组织化学染色。采用酶联免疫吸附法检测犬关节液中IL-1 Ra水平。通过聚合酶链反应检测滑膜中DNA质粒的存在。观察到骨赘宽度和肉眼可见病变大小的显著减小（P < 0.04），并且依赖于注射的IL-IRa质粒的量。在接受最高剂量（1000 Erg）IL-1 Ra质粒的组中，组织学软骨损伤的严重程度也显著降低（CP < 0.01）。通过酶联免疫吸附测定和免疫组织化学染色在接受含有IL-1 Ra质粒的注射的兔的滑膜和软骨中检测到滑液中的IL-1 Ra。滑膜DNA的聚合酶链反应分析揭示了克隆的cDNA犬IL-1 Ra的存在下，第一次关节内注射后4周。这项研究表明，使用质粒载体和脂质的关节内注射将IL-1 Ra基因直接体内转移到骨关节炎膝关节细胞中可以显著降低实验性骨关节炎的进展。因此，这种途径可能代表了骨关节炎有前途的未来治疗方法。
The goal of this study was to determine the efficacy of local IL-1Ra gene therapy by intra-articular plasmid injections on structural changes in the meniscectomy rabbit model of osteoarthritis. A partial meniscectomy of the right knee was performed on the rabbits through a medial parapatellar incision. The rabbits were then divided into four experimental groups. Group 1 received no treatment. Group 2 received three consecutive intra-articular injections at 24-hour intervals of 0.9% saline containing a lipid, gamma AP-DL-RIE/DOPE, and a DNA plasmid, VR1012. Group 3 received three consecutive injections of saline containing 1000 mu g of canine IL-1Ra plasmid and lipid. The injections were given starting 4 weeks post-surgery. Rabbits from Group 1 were killed 4 weeks post-surgery, and all other rabbits 8 weeks post-surgery, The severity of macroscopic and microscopic changes on cartilage on the medial and femoral condyles and tibial plateaus and synovium were graded separately. Specimens were also processed for immunohistochemical staining using a rabbit polyclonal antibody against canine IL-1Ra. The level of canine IL-1Ra in synovial fluid was determined using enzyme-linked immunosorbent assay. The presence of the DNA plasmid in the synovium was tested by polymerase chain reaction. A significant reduction in the width of osteophytes and size of macroscopic lesions (P < 0.04) was observed, and was dependent on the amount of IL-IRa plasmid injected. A significant reduction was also noted in the severity of histologic cartilage lesions CP < 0.01) in the group that received the highest dosage (1000 Erg) of IL-1Ra plasmid, IL-1Ra was detected in synovial fluid by enzyme-linked immunosorbent assay and by immunohistochemical staining in the synovium and cartilage of rabbits that received injections containing the IL-IRa plasmid. Polymerase chain reaction analysis of synovial DNA revealed the presence of the cloned cDNA dog IL-1Ra up to 4 weeks after the first intra-articular injection. This study demonstrates that direct in vivo transfer of the IL-1Ra gene into osteoarthritis knee cells using intra-articular injections of a plasmid vector and lipids can significantly reduce the progression of experimental osteoarthritis. This avenue may therefore represent a promising future treatment for osteoarthritis.