In vivo molecular imaging of adenoviral versus lentiviral gene therapy in two bone formation models
In vivo molecular imaging of adenoviral versus lentiviral gene therapy in two bone formation models
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DOI：
10.1002/jor.20229
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发表时间：
2006-08-01
影响因子：
2.8
通讯作者：
Lieberman, Jay R.
中科院分区：
文献类型：
--
作者：
Feeley, Brian T.;Conduah, Augustine H.;Lieberman, Jay R.
区域性基因治疗技术是一种很有前途的方法，可以促进同种异体骨或自体骨难以治疗的大面积骨缺损的骨形成。在这项研究中，我们比较了腺病毒和慢病毒介导的基因治疗在两种骨形成模型中的体内时间表达模式。用含有荧光素酶（Luc）或BMP-2 cDNA的慢病毒或腺病毒载体转导原代大鼠骨髓细胞（RBMC），或与含有Luc和骨形态发生蛋白2（BMP-2）的载体共转导。用荧光素酶测定或ELISA（用于BMP-2产生）每周测定体外蛋白质产生，持续12周。在SCID小鼠中使用了两种骨形成模型-后肢肌袋或桡骨缺损。使用冷却的电荷耦合器件（CCD）照相机每周对体内荧光素酶表达成像，持续12周。在体外，BMP-2和荧光素酶的腺病毒表达分别通过ELISA或荧光素酶测定法检测，持续4周。BMP-2和荧光素酶的慢病毒表达在培养物中持续3个月。使用CCD摄像机，我们发现腺病毒载体表达荧光素酶表达长达21天，但慢病毒载体表达靶基因表达3个月在两种骨形成模型体内。在腺病毒组和慢病毒组之间，骨形成量没有可检测到的差异。BMP-2的慢病毒介导的递送可以诱导长期的体外和体内基因表达，这在开发组织工程策略以愈合大的骨缺损或具有受损生物环境的缺损时可能是有益的。(c)2006骨科研究学会。出版社：Wiley Periodicals，Inc.
Regional gene therapy techniques are promising methods to enhance bone formation in large bone defects that would be difficult to treat with allograft or autograft bone stock. In this study, we compared in vivo temporal expression patterns of adenoviral- and lentiviral-mediated gene therapy in two bone formation models. Primary rat bone marrow cells (RBMC) were transduced with lentiviral or adenoviral vectors containing luciferase (Luc) or BMP-2 cDNA, or cotransduced with vectors containing Luc and bone morphogenetic protein 2 (BMP-2). In vitro protein production was determined with luciferase assay or ELISA (for BMP-2 production) weekly for 12 weeks. Two bone formation models were used-a hind limb muscle pouch or radial defect-in SCID mice. A cooled charged-coupled device (CCD) camera was used to image in vivo luciferase expression weekly for 12 weeks. In vitro, adenoviral expression of BMP-2 and luciferase was detected by ELISA or luciferase assay, respectively, for 4 weeks. Lentiviral expression of BMP-2 and luciferase was sustained in culture for 3 months. Using the CCD camera, we found that adenoviral vectors expressed luciferase expression for up to 21 days, but lentiviral vectors expressed target gene expression for 3 months in vivo in both bone formation models. There was no detectable difference in the amount of bone formed between the adenoviral and lentiviral groups. Lentiviral-mediated delivery of BMP-2 can induce long term in vitro and in vivo gene expression, which may be beneficial when developing tissue engineering strategies to heal large bone defects or defects with a compromised biologic environment. (c) 2006 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.