van Griensven M, Lobenhoffer P, Barke A, Tschernig T, Lindenmaier W, Krettek C, Gerich T G
Department of Trauma Surgery, Hannover Medical School, Carl-Neuberg-Strasse 1, D-30625, Hannover, Germany.
Lab Anim. 2002 Oct;36(4):455-61. doi: 10.1258/002367702320389134.
For the enhancement of fracture healing, either purified proteins or vectors for expression of growth factors in situ may be used. Adenoviral vectors directly convert cells to express a transgene. However, the cell types which are preferentially infected and the time of expression during fracture healing are currently not known. The adenoviral type 5 vectors used in this study are replication incompetent viruses, one encoding beta-galactosidase (beta-GAL) and one green fluorescent protein. Femora of 35 Sprague-Dawley rats were fractured. Three days after stabilization with Kirschner wire, 10(12) pfu viral suspension were injected into the fracture zone. As a control, five animals received injections of adenovirus type 2. Animals were sacrificed after 3 days, 1, 2 and 4 weeks. Fractures healed radiographically within 2-3 weeks. All specimens were examined for beta-GAL and green fluorescent protein (GFP) expression. Fibroblast and osteoblasts within callus tissue displayed a high transgene expression (week 1). A decrease of expression was observed during the observation period. In this experimental study, we have demonstrated that all cells of the primary callus can be transfected using adenoviral vectors, which provide a tool to further investigate adenoviral transfer of growth factors such as bone morphogenetic protein-2 (BMP-2).
为促进骨折愈合，可使用纯化蛋白或原位表达生长因子的载体。腺病毒载体可直接促使细胞表达转基因。然而，目前尚不清楚骨折愈合过程中哪些细胞类型会被优先感染以及表达时间。本研究中使用的5型腺病毒载体是无复制能力的病毒，一种编码β-半乳糖苷酶（β-GAL），另一种编码绿色荧光蛋白。对35只Sprague-Dawley大鼠的股骨进行骨折。用克氏针固定3天后，将10(12) pfu病毒悬液注入骨折区域。作为对照，5只动物注射2型腺病毒。在3天、1周、2周和4周后处死动物。骨折在2至3周内通过X线检查愈合。对所有标本进行β-GAL和绿色荧光蛋白（GFP）表达检测。骨痂组织中的成纤维细胞和成骨细胞显示出高转基因表达（第1周）。在观察期内观察到表达下降。在本实验研究中，我们证明了使用腺病毒载体可以转染原始骨痂的所有细胞，这为进一步研究骨形态发生蛋白-2（BMP-2）等生长因子的腺病毒转移提供了一种工具。
