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Interaction of keratinocytes and fibroblasts modulates the expression of matrix metalloproteinases-2 and -9 and their inhibitors

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Abstract

Disruption of epidermal-mesenchymal communication due to a delay in epithelialization, increases the frequency of developing fibrotic conditions in skin. As matrix metalloproteinases-2 (MMP-2) and -9 (MMP-9) are two key enzymes involved in wound healing and tissue remodeling, here we examined the efficacy of keratinocyte-fibroblast interaction on modulation of these enzymes and their inhibitors. The conditioned media derived from keratinocytes and fibroblasts grown in upper and lower chambers of a co-culture system, respectively, were analyzed for MMP-2 and -9. Keratinocyte or fibroblast conditioned medium (FCM) was used as a control. Gelatinolytic activity analyzed by zymography showed that keratinocytes mainly express MMP-9 and to a lesser extent MMP-2; while fibroblasts express only MMP-2. In a co-culture system, the activities of both MMP-2 and MMP-9 markedly increased in conditioned media collected from bottom chambers. These findings were consistent with the level of MMP-2 and MMP-9 measured by Western blot. Using the same experimental setting, the levels of tissue inhibitors of MMPs (TIMPs) secreted by keratinocytes and fibroblasts grown in the same co-culture system were also evaluated. Western blot showed that fibroblasts secrete only TIMP-1 and TIMP-2 whose levels were increased by co-culturing fibroblasts with keratinocytes. In contrary the level of TIMP-3, which was mainly expressed by keratinocytes, increased by co-culturing these cells with fibroblasts. In conclusion, interaction of fibroblast-keratinocyte modulates the levels of MMP-2 and -9 and their inhibitors produced by these cells and this interaction may be critical for a better healing quality at a late stage of the wound healing process. (Mol Cell Biochem 269: 209–216, 2005)

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Authors and Affiliations

  1. Department of Pharmacology, Cardiovascular Research Group, University of Alberta, Edmonton, Alberta, Canada

    Grzegorz Sawicki

  2. Department of Surgery, Wound Healing Research Group, University of Alberta, 161 HMRC, Edmonton, Canada

    Yvonne Marcoux, Kourosh Sarkhosh, Edward E. Tredget & Aziz Ghahary

Authors
  1. Grzegorz Sawicki
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  2. Yvonne Marcoux
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  3. Kourosh Sarkhosh
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  4. Edward E. Tredget
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  5. Aziz Ghahary
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Correspondence to Aziz Ghahary.

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Sawicki, G., Marcoux, Y., Sarkhosh, K. et al. Interaction of keratinocytes and fibroblasts modulates the expression of matrix metalloproteinases-2 and -9 and their inhibitors. Mol Cell Biochem 269, 209–216 (2005). https://doi.org/10.1007/s11010-005-3178-x

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  • DOI: https://doi.org/10.1007/s11010-005-3178-x

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