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Effect of transfection strategy on growth factor overexpression by articular chondrocytes

Articular cartilage damage remains an unsolved problem in orthopaedics. Insulin‐like growth factor I (IGF‐I) and fibroblast growth factor‐2 (FGF‐2) are anabolic and mitogenic for articular chondrocytes, and are candidates for the application of gene therapy to articular cartilage repair. We tested the hypothesis that the production of IGF‐I and FGF‐2 can be augmented by modulating vector designs and delivery methods used for gene transfer to articular chondrocytes. We developed a novel adeno‐associated virus (AAV)‐based plasmid (pAAV) to overexpress IGF‐I and FGF‐2 cDNAs in adult bovine articular chondrocytes. We found that the pAAV‐based vectors generated significantly more growth factor than pcDNA vectors carrying the same cDNAs. Chondrocytes cotransfected with both IGF‐I and FGF‐2 cDNAs in two separate pAAV plasmids produced significantly more IGF‐I and FGF‐2 than cells transfected by a single pAAV plasmid carrying both cDNAs in a dicistronic cassette. These data indicate that pAAV vectors are more effective than pcDNA vectors for transfer of IGF‐I and FGF‐2 genes to articular chondrocytes. They further suggest that cotransfection may be an effective strategy for multiple gene transfer to these cells. These findings may be important in applying growth factor gene transfer to cell‐based articular cartilage gene therapy. © 2009 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 28:103–109, 2010