Baltzer A W, Lattermann C, Whalen J D, Braunstein S, Robbins P D, Evans C H
Orthopädische Universitätsklinik, Heinrich-Heine-Universität, Düsseldorf, Germany.
Knee Surg Sports Traumatol Arthrosc. 1999;7(3):197-202. doi: 10.1007/s001670050147.
It has been demonstrated that BMPs, IGFs, and TGFbetas improve the process of bone healing in vivo. We have suggested the use of gene therapy as a possible way to deliver growth factors to fracture sites in order to improve repair. The aim of this study was to develop a minimally invasive gene therapy approach to treat bone injuries locally without damaging the local blood circulation. A segmental defect of 1.3 cm was created in the diaphysis of the femur in mature NZW rabbits. Internal fixation with 7-hole DCP plates and 2.7 mm screws was used to stabilize the bone. After building a chamber by tightly closing the muscles around the segmental defect, 0.5 ml of either saline solution or a collagen gel containing 1 x 10(10) particles of adenovirus carrying cDNA encoding either the bacterial beta-galactosidase gene (LacZ), or the firefly luciferase gene were injected into the gap. The control side received 0.5 ml of saline solution without virus particles. Bone marrow, cortical and trabecular bone and surrounding muscle were harvested from the injected femur and were analyzed for local gene expression through X-gal staining or measurement of local luciferase activity. To determine whether distant sites were transduced, tissue from the spleen, liver, and lung were harvested as well as bone, bone marrow and muscle from the contralateral diaphysis of the femur. The delivery of the adenoviral vector suspended in saline solution led to local transduction of the bone, bone marrow and the muscle surrounding the gap. No luciferase activity was found in the contralateral femur, lung, or spleen, and only transient luciferase activity was seen in the liver. While marker gene expression persisted within the surrounding soft tissues for at least 2 weeks, the expression in bone lasted up to 6 weeks. This study has shown that it is possible to use adenoviral vectors to transfer and express genes locally within a segmental defect. Gene expression persisted for several weeks, which may be already sufficient to accelerate repair.
已有研究表明，骨形态发生蛋白（BMPs）、胰岛素样生长因子（IGFs）和转化生长因子β（TGFbetas）可改善体内骨愈合过程。我们曾提出，基因治疗可能是一种将生长因子输送至骨折部位以促进修复的方法。本研究的目的是开发一种微创基因治疗方法，在不损害局部血液循环的情况下局部治疗骨损伤。在成年新西兰白兔的股骨干中制造一个1.3厘米的节段性骨缺损。使用7孔动力加压接骨板（DCP）和2.7毫米螺钉进行内固定以稳定骨骼。通过紧密闭合节段性缺损周围的肌肉构建一个腔室后，将0.5毫升生理盐水或含有1×10（10）个携带编码细菌β-半乳糖苷酶基因（LacZ）或萤火虫荧光素酶基因的腺病毒颗粒的胶原凝胶注入间隙中。对照侧注射0.5毫升不含病毒颗粒的生理盐水。从注射的股骨中采集骨髓、皮质骨、小梁骨和周围肌肉，并通过X-gal染色或测量局部荧光素酶活性分析局部基因表达。为了确定远处部位是否被转导，还采集了脾脏、肝脏和肺的组织以及对侧股骨干的骨、骨髓和肌肉。悬浮于生理盐水溶液中的腺病毒载体的递送导致间隙周围的骨、骨髓和肌肉发生局部转导。在对侧股骨、肺或脾脏中未发现荧光素酶活性，仅在肝脏中观察到短暂的荧光素酶活性。虽然标记基因在周围软组织中的表达持续至少2周，但在骨中的表达持续长达6周。本研究表明，使用腺病毒载体在节段性缺损内局部转移和表达基因是可行的。基因表达持续数周，这可能已足以加速修复。
