The First Type III Repeat in Fibronectin Activates an Inflammatory Pathway in Dermal Fibroblasts

Remodeling of the fibronectin matrix occurs during a variety of pathological and regenerative processes. Cellular generated tensional forces can alter the secondary and tertiary structure of the fibronectin matrix and regulate the exposure of cryptic activities that directly impact cell behavior. In the present study, we evaluated the effect of the partially unfolded Type III fibronectin module, FnIII-1c, on gene expression in dermal fibroblasts. Microarray and PCR analysis indicated that the addition of FnIII-1c to human dermal fibroblasts induced the expression of several inflammatory genes including the cytokines, IL-8 and TNF-alpha. ELISA analysis indicated that the increased gene expression was accompanied by the secretion of IL-8 and TNF-alpha protein. FnIII-1c-induced gene expression was preceded by increased phosphorylation of I kappa B kinase (IKK) and I kappa B alpha as well as the nuclear translocation of NF kappa B. PCR and ELISA analysis showed that inhibition of the NF kappa B signaling pathway completely blocked the induction of IL-8 and TNF-alpha. Blocking antibodies to Toll-like receptor 4 inhibited both the activation of the NF kappa B signaling pathway as well as cytokine expression in response to FnIII-1c. These data suggest that fibronectin matrix remodeling can induce the expression of cytokines by stromal cells present in the tissue microenvironment.