Genetic modification of chondrocytes with insulin-like growth factor-1 enhances cartilage healing in an equine model
Genetic modification of chondrocytes with insulin-like growth factor-1 enhances cartilage healing in an equine model
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DOI：
10.1302/0301-620x.89b5.18343
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发表时间：
2007-05-01
影响因子：
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通讯作者：
Nixon, A. J.
中科院分区：
文献类型：
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作者：
Goodrich, L. R.;Hidaka, C.;Nixon, A. J.
用胰岛素样生长因子-1 （IGF-1）进行基因治疗可以增加基质的产生，增强软骨细胞的增殖和体外存活。本研究的目的是确定经编码马IGF-1 （AdIGF-1）的腺病毒载体基因修饰的关节镜移植软骨细胞是否对马股髌骨关节模型的软骨愈合有有益的影响。共有16匹马接受了每个股髌骨关节单个15毫米软骨缺损的关节镜修复。一侧关节接受2 × 107个AdIGF-1修饰的软骨细胞，对侧关节接受2 × 107个未修饰的软骨细胞。术后4周、9周和8个月分别对修复情况进行分析。形态学和组织学外观、IGF-1和II型胶原基因表达（聚合酶链反应、原位杂交和免疫组织化学）、II型胶原含量（溴化氰和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳）、蛋白聚糖含量（二甲基亚甲基蓝测定）、I型胶原、基质金属蛋白酶(MMP)-1、MMP-3、MMP-13、聚合酶-1、检测基质金属蛋白酶-1 （TIMP-1）和TIMP-3的组织抑制因子。在移植后长达9周的时间里，软骨细胞的遗传修饰显著增加了修复组织中IGF-1 mRNA和配体的产生。IGF-1修饰的修复组织的大体和组织学外观比对照缺陷改善。缺损的肉眼充盈在四周时得到显著改善，在8个月时病变被更透明样的组织覆盖。移植后4周和9周的组织学结果显示，转基因软骨细胞移植后的缺损组织填充更大，而对照缺损的修复组织更薄、不规则且纤维性更强。在IGF-1基因转导的缺陷中，II型胶原蛋白的表达在4周时增加了100倍，并且与11型胶原蛋白免疫反应的增加相关，直至8个月。在移植前用AdIGF-1对软骨细胞进行遗传修饰，可在早期（4至9周）改善马模型的软骨愈合，并在较小程度上改善长期软骨愈合。马软骨愈合模型与人类临床软骨修复非常相似。本研究结果表明，在移植前对软骨细胞进行基因修饰可以促进软骨愈合。
Gene therapy with insulin-like growth factor-1 (IGF-1) increases matrix production and enhances chondrocyte proliferation and survival in vitro. The purpose of this study was to determine whether arthroscopically-grafted chondrocytes genetically modified by an adenovirus vector encoding equine IGF-1 (AdIGF-1) would have a beneficial effect on cartilage healing in an equine femoropatellar joint model.A total of 16 horses underwent arthroscopic repair of a single 15 mm cartilage defect in each femoropatellar joint. One joint received 2 x 107 AdIGF-1 modified chondrocytes and the contralateral joint received 2 x 107 naive (unmodified) chondrocytes. Repairs were analysed at four weeks, nine weeks and eight months after surgery. Morphological and histological appearance, IGF-1 and collagen type II gene expression (polymerase chain reaction, in situ hybridisation and immunohistochemistry), collagen type II content (cyanogen bromide and sodium dodecyl sulphate-polyacrylamide gel electrophoresis), proteoglycan content (dimethylmethylene blue assay), and gene expression for collagen type I, matrix metalloproteinase (MMP)-1, MMP-3, MMP-13, aggrecanase-1, tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) and TIMP-3 were evaluated.Genetic modification of chondrocytes significantly increased IGF-1 mRNA and ligand production in repair tissue for up to nine weeks following transplantation. The gross and histological appearance of IGF-1 modified repair tissue was improved over control defects. Gross filling of defects was significantly improved at four weeks, and a more hyaline-like tissue covered the lesions at eight months. Histological outcome at four and nine weeks post-transplantation revealed greater tissue filling of defects transplanted with genetically modified chondrocytes, whereas repair tissue in control defects was thin and irregular and more fibrous. Collagen type II expression in IGF-1 gene-transduced defects was increased 100-fold at four weeks and correlated with increased collagen type 11 immunoreaction up to eight months.Genetic modification of chondrocytes with AdIGF-1 prior to transplantation improved early (four to nine weeks), and to a lesser degree long-term, cartilage healing in the equine model.The equine model of cartilage healing closely resembles human clinical cartilage repair. The results of this study suggest that cartilage healing can be enhanced through genetic modification of chondrocytes prior to transplantation.