Adeno-associated virus-mediated osteoprotegerin gene transfer protects against particulate polyethylene-induced osteolysis in a murine model
Adeno-associated virus-mediated osteoprotegerin gene transfer protects against particulate polyethylene-induced osteolysis in a murine model
复制标题
DOI：
10.1002/art.10527
复制
发表时间：
2002-09-01
影响因子：
--
通讯作者：
Wooley, PH
中科院分区：
文献类型：
--
作者：
Yang, SY;Mayton, L;Wooley, PH
目标。护骨素(OPG)是破骨细胞生成和骨吸收的天然负性调节因子，可能成为治疗溶骨性假体关节松动的潜在药物。采用在体腺相关病毒(AAV)介导的基因转移技术，在小鼠骨溶解模型中评价转基因OPG对骨科磨屑所致骨丢失的保护作用。将骨组织植入BALB/c小鼠已建立的骨囊中，然后引入超高相对分子质量聚乙烯(UHMWPE)颗粒，以刺激炎症和骨溶解。将编码人OPG基因的病毒(rAAV-hOPG)或β-半乳糖苷酶标志基因(rAAV-lacZ)注入气囊，感染后7d取材进行组织学和分子生物学分析。通过酶联免疫吸附试验检测OPG和眼袋组织X-Gal染色(LacZ)，证实转基因成功表达。实时定量聚合酶链式反应显示，与rAAV-LacZ基因转导的骨囊相比，OPG转导的破骨细胞标志物的信使RNA表达显著减少。OPG的转导和表达也显著降低了核因子kappaB生物受体激活物的基因拷贝。与其他两组相比，植入骨囊中OPG的表达平均减少了39%的骨钙释放。计算机图像分析显示，OPG的表达对骨胶原丢失有明显的保护作用。在本实验模型中，rAAV介导的OPG基因转移有效地保护了颗粒聚乙烯诱导的骨吸收。数据表明，使用rAAV-OPG的基因转移可能是治疗或防止磨屑相关的骨溶解和无菌性松动的一种可行和有效的治疗方案。
Objective. Osteoprotegerin (OPG), a natural negative regulator of osteoclastogenesis and bone resorption, may be a potential therapeutic agent for treatment of osteolysis-associated prosthetic joint loosening. Using an in vivo adeno-associated virus (AAV)-mediated gene transfer technique, this study was designed to evaluate the protective effects of OPG transgene against orthopedic wear debris-induced bone loss in a murine model of osteolysis.Methods. Bone tissue was implanted into established pouches on BALB/c mice, followed by the introduction of ultra-high-molecular-weight polyethylene (UHMWPE) particles to provoke inflammation and osteolysis. The viruses encoding human OPG gene (rAAV-hOPG) or beta-galactosidase marker gene (rAAV-LacZ) were injected into the air pouches, and the tissue was harvested 7 days after viral infection for histologic and molecular analyses.Results. Successful transgene expression was confirmed by the detection of OPG by enzyme-linked immunosorbent assay and positive X-Gal staining of pouch tissue (LacZ). Real-time polymerase chain reaction indicated significant diminishment of messenger RNA expression of osteoclast markers in OPG-transduced pouches compared with rAAV-LacZ-transduced pouches. The transduction and expression of OPG also markedly decreased the gene copies of the biologic receptor activator of nuclear factor kappaB. The expression of OPG in the bone-implanted pouch reduced bone calcium release by a mean of 39% compared with the calcium release in the other 2 groups. Computerized image analysis revealed that expression of OPG significantly protected against bone collagen loss.Conclusion. OPG gene transfer mediated by rAAV effectively protects against particulate polyethylene-induced bone resorption in this experimental model. Data suggest that gene transfer using rAAV-OPG may be a feasible and effective therapeutic candidate to treat or prevent wear debris-associated osteolysis and aseptic loosening.