Detectable Reporter Gene Expression following Transduction of Adenovirus and Adeno-Associated Virus Serotype 2 Vectors within Full-Thickness Osteoarthritic and Unaffected Canine Cartilage In Vitro and Unaffected Guinea Pig Cartilage In Vivo
Detectable Reporter Gene Expression following Transduction of Adenovirus and Adeno-Associated Virus Serotype 2 Vectors within Full-Thickness Osteoarthritic and Unaffected Canine Cartilage In Vitro and Unaffected Guinea Pig Cartilage In Vivo
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DOI：
10.1002/jor.20975
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发表时间：
2010-02-01
影响因子：
2.8
通讯作者：
Bertone, Alicia L.
中科院分区：
文献类型：
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作者：
Santangelo, Kelly S.;Baker, Sarah A.;Bertone, Alicia L.
本研究定量并比较了腺病毒（Ad）、Arg-Gly-Asp（RGD）修饰的Ad、腺相关病毒血清型2（AAV 2）和自身互补AAV 2（scAAV 2）载体在体外全层骨关节炎（OA）和未受影响的犬软骨外植体中的转导效率。进行Ad和scAAV 2载体的关节内施用以确定这些载体在体内活化未受影响的豚鼠软骨的能力。在外植体暴露于载体处理或对照后，每天用荧光显微镜监测报告基因表达的开始和表面分布。在处死时，将外植体分开：一半消化用于使用流式细胞术分析;剩余部分用于组织学和免疫组织化学（IHC）。收集完整的关节进行实时RT-PCR和IHC，以检测注射所选载体后报告基因的表达。Ad载体沿着外植体的周边沿着转导病灶区域;剩余的载体在100%的表面上转导软骨细胞。与Ad载体相比，发现两种AAV 2载体的平均转导效率更高（p
This study quantified and compared the transduction efficiencies of adenoviral (Ad), Arg-Gly-Asp (RGD)-modified Ad, adeno-associated viral serotype 2 (AAV2), and self-complementary AAV2 (scAAV2) vectors within full-thickness osteoarthritic (OA) and unaffected canine cartilage explants in vitro. Intraarticular administration of Ad and scAAV2 vectors was performed to determine the ability of these vectors to transduce unaffected guinea pig cartilage in vivo. Following explant exposure to vector treatment or control, the onset and surface distribution of reporter gene expression was monitored daily with fluorescent microscopy. At termination, explants were divided: one half was digested for analysis using flow cytometry; the remaining portion was used for histology and immunohistochemistry (IHC). Intact articular joints were collected for real-time RT-PCR and IHC to detect reporter gene expression following injection of selected vectors. Ad vector transduced focal areas along the perimeters of explants; the remaining vectors transduced chondrocytes across 100% of the surface. Greater mean transduction efficiencies were found with both AAV2 vectors as compared to the Ad vector (p