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Thromb Haemost 2004; 92(02): 262-274
DOI: 10.1160/TH03-11-0669
Theme Issue Article
Schattauer GmbH

Dissecting the roles of endothelin,TGF-β and GM-CSF on myofibroblast differentiation by keratinocytes

Pierre Shephard
1   Department of Dermatology, University of Cologne, Cologne, Germany
,
Boris Hinz
2   Laboratory of Cell Biophysics, Swiss Federal Institute of Technology (EPFL), Lausanne, Switzerland
,
Sigrun Smola-Hess
3   Department of Virology, University of Cologne, Cologne, Germany
,
Jean-Jacques Meister
2   Laboratory of Cell Biophysics, Swiss Federal Institute of Technology (EPFL), Lausanne, Switzerland
,
Thomas Krieg
1   Department of Dermatology, University of Cologne, Cologne, Germany
,
Hans Smola   §
1   Department of Dermatology, University of Cologne, Cologne, Germany
› Author AffiliationsFinancial support: This work was supported by grants from the Deutsche Forschungsgemeinschaft Kr558/13-1, SFB 589.S (DFG), Germany, the Center of Molecular Medicine Cologne (ZMMK), Germany and by the Swiss National Science Foundation, grant #3100A0-102150/1 to B.H.. S.S-H. received a Heisenberg Stipendium from the Deutsche Forschungsgemeinschaft (DFG), Germany.
Further Information

Publication History

Received 04 November 2003

Accepted after resubmission 01 June 2004

Publication Date:
04 December 2017 (online)

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Summary

Myofibroblasts are specialized fibroblasts that contribute to wound healing by producing extracellular matrix and by contracting the granulation tissue.They appear in a phase of wound healing when the dermis strongly interacts with activated epidermal keratinocytes. Direct co-culture with keratinocytes upregulates TGF-β???????????activity and also induces fibroblast to differentiate into α-smooth muscle actin (αSMA)-positive myofibroblasts. TGF-βactivity alone cannot completely account for αSMA induction in these co-cultures, and here we analyze mechanical force generation, another potent inducer of myofibroblast differentiation in this model. Using deformable silicone substrates, we show that contractile activity of fibroblasts is already induced after 1-2-days of co-culture, when fibroblasts are generally αSMA negative. Endothelin-1 (ET-1), the most potent inducer of smooth muscle cell contraction, was up-regulated in co-cultures, while blocking ET-1 with the ET receptor inhibitor PD156252 inhibited contraction in these early co-cultures. In 4-5 days of co-culture, however, fibroblast contractile activity correlated with an increased expression of αSMA expression. Stimulation of fibroblast mono-cultures with ET-1 in a low serum medium did not induce αSMA expression; however, ET-1 did synergize with TGF-β. Surprisingly, GM-CSF, another mediatorstimulating myofibroblast differentiation in granulation tissue, inhibited αSMA expression in fibroblasts, costimulated with TGF-β and ET-1. GM-CSF activated NFκB, thus interfering with TGF-β signaling. Blocking TGF-β and ET-1 largely impaired αSMA induction in co-cultures at day 7 and, in combination, almost completely prevented αSMA induction. Our results dissect the roles of TGF-β and ET-1 on mechanical force generation in keratinocyte-fibroblast co-cultures, and identify GM-CSF as an inducer of myofibroblasts acting indirectly.

Keywords

Wound healing - myofibroblasts - endothelin,TGF-β - GM-CSF

§ Current address: Nestlé Research Center, Lausanne, Switzerland


 

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