Background: Gene therapy has the potential to provide long-term production of therapeutic proteins in the joints of\nosteoarthritis (OA) patients. The objective of this study was to analyse the therapeutic potential of disease-inducible\nexpression of anti-inflammatory interleukin-10 (IL-10) in the three-dimensional micromass model of the human\nsynovial membrane.\nMethods: Synovial tissue samples from OA patients were digested and the cells were mixed with Matrigel to obtain\n3D micromasses. The CXCL10 promoter combined with the firefly luciferase reporter in a lentiviral vector was used to\ndetermine the response of the CXCL10 promoter to tumour necrosis factor alpha (TNF-�±), interleukin-1�² (IL-1�²) and\nlipopolysaccharide (LPS). The effects of recombinant IL-10 on gene expression were determined by quantitative PCR.\nThe production of IL-10 from the CXCL10p-IL10 vector and the effects on pro-inflammatory cytokine production were\nassessed by multiplex ELISA.\nResults: Micromasses made from whole synovial membrane cell suspensions form a distinct surface composition\ncontaining macrophage and fibroblast-like synoviocytes thus mimicking the synovial lining. This lining can be\ntransduced by lentiviruses and allow CXCL-10 promoter-regulated transgene expression. Adequate amounts of IL-10\ntransgene were produced after stimulation with pro-inflammatory factors able to reduce the production of synovial\nIL-1�² and IL-6.\nConclusions: Synovial micromasses are a suitable model to test disease-regulated gene therapy approaches and\nthe CXCL10p-IL10 vector might be a good candidate to decrease the inflammatory response implicated in the\npathogenesis of OA.
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