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Microarray Analysis of Gene Expression Reveals that Cyclo-oxygenase-2 Gene Therapy Up-regulates Hematopoiesis and Down-regulates Inflammation During Endochondral Bone Fracture Healing

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Background: Cyclo-oxygenase-2 (Cox-2) is an inflammatory mediator that is necessaryfor the tissue repair, including bone fracture healing. Although the application of Cox-2gene therapy to a murine closed femoral fracture has accelerated bony union, but thebeneficial effect was not observed until the endochondral stage of bone repair that iswell after the inflammatory stage normally subsides. Methods: To identify the molecularpathways through which Cox-2 regulates fracture healing, we examined gene expres-sion profile in fracture tissues in response to Cox-2 gene therapy during the endochon-dral bone repair phase. Cox-2 gene therapy was applied to the closed murine femurfracture model. Microarray analysis was performed at 10 days post-fracture to examineglobal gene expression profile in the fracture tissues during the endochondral bone re-pair phase. The entire repertoire of significantly expressed genes was examined by geneset enrichment analysis, and the most up-regulated individual genes were evaluatedfurther. Results: The genes that normally promote inflammation were under-represent-ed in the microarray analysis, and the expression of several inflammatory chemokineswas significantly down-regulated. There was an up-regulation of two key transcriptionfactor genes that regulate hematopoiesis and erythropoiesis. More surprisingly, therewas no significant up-regulation in the genes that are normally involved in angiogenesisor bone formation. However, the expression of two tissue remodeling genes was up-regulated. Conclusions: The down-regulation of the inflammatory genes in response toCox-2 gene therapy was unexpected, given the pro-inflammatory role of prostaglandins. Cox-2 gene therapy could promote bony union through hematopoietic precursor prolif-eration during endochondral bone repair and thereby enhances subsequently fracturecallus remo deling that leads to bony union of the fracture gap.

Abstract

Background: Cyclo-oxygenase-2 (Cox-2) is an inflammatory mediator that is necessaryfor the tissue repair, including bone fracture healing. Although the application of Cox-2gene therapy to a murine closed femoral fracture has accelerated bony union, but thebeneficial effect was not observed until the endochondral stage of bone repair that iswell after the inflammatory stage normally subsides. Methods: To identify the molecularpathways through which Cox-2 regulates fracture healing, we examined gene expres-sion profile in fracture tissues in response to Cox-2 gene therapy during the endochon-dral bone repair phase. Cox-2 gene therapy was applied to the closed murine femurfracture model. Microarray analysis was performed at 10 days post-fracture to examineglobal gene expression profile in the fracture tissues during the endochondral bone re-pair phase. The entire repertoire of significantly expressed genes was examined by geneset enrichment analysis, and the most up-regulated individual genes were evaluatedfurther. Results: The genes that normally promote inflammation were under-represent-ed in the microarray analysis, and the expression of several inflammatory chemokineswas significantly down-regulated. There was an up-regulation of two key transcriptionfactor genes that regulate hematopoiesis and erythropoiesis. More surprisingly, therewas no significant up-regulation in the genes that are normally involved in angiogenesisor bone formation. However, the expression of two tissue remodeling genes was up-regulated. Conclusions: The down-regulation of the inflammatory genes in response toCox-2 gene therapy was unexpected, given the pro-inflammatory role of prostaglandins. Cox-2 gene therapy could promote bony union through hematopoietic precursor prolif-eration during endochondral bone repair and thereby enhances subsequently fracturecallus remo deling that leads to bony union of the fracture gap.

참고문헌 (45)

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