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TGF-β1 regulates TGF-β1 and FGF-2 mRNA expression during fibroblast wound healing
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  1. Q H Song3,
  2. V E Klepeis2,
  3. M A Nugent1,
  4. V Trinkaus-Randall1
  1. 1Department of Ophthalmology, Boston University School of Medicine, Boston, MA 02118, USA
  2. 2Department of Pathology, Boston University School of Medicine
  3. 3Department of Biochemistry, Boston University School of Medicine
  1. Correspondence to:
 Dr V Trinkaus-Randall, Boston University School of Medicine, L903, 80 East Concord Street, Boston, MA 02118, USA;
 vickery{at}biochem.bumc.bu.edu

Abstract

Aims: To evaluate the expression of transforming growth factor β1 (TGF-β1) and fibroblast growth factor 2 (FGF-2) mRNA in stromal cells in response to injury in the presence of either TGF-β1 or FGF-2. It has been shown previously that heparan sulfate proteoglycans and FGF-2 are present transiently during wound repair in vivo and that an increase in TGF-β1 mRNA is detected rapidly after injury.

Methods: Primary corneal fibroblasts were cultured to confluency, serum starved, and linear wound(s) were made in medium containing TGF-β1 or FGF-2. TGF-β1 and FGF-2 mRNA expression were evaluated using both northern blot analysis and in situ hybridisation. Both dose dependent and time course experiments were performed. Whole eye organ culture experiments were also carried out and growth factor expression was assessed.

Results: Injury and exogenous TGF-β1 increased TGF-β1 mRNA values. The increase in expression of FGF-2 mRNA was not detected until wound closure. In contrast, FGF-2 inhibited the expression of TGF-β1. TGF-β1 increased TGF-β1 mRNA stability but did not alter that of FGF-2. Migration assay data demonstrated that unstimulated stromal cells could be activated to migrate to specific growth factors.

Conclusions: TGF-β1 specifically enhances cellular responsiveness, as shown by increased stability after injury and the acquisition of a migratory phenotype. These data suggest that there is an integral relation during wound repair between TGF-β1 and FGF-2.

  • wound repair
  • migration
  • transforming growth factor β1
  • fibroblast growth factor 2
  • BSA, bovine serum albumin
  • DRB, 5,6-dichlorobenzmidazole riboside
  • FBS, fetal bovine serum
  • FGF-2, fibroblast growth factor 2
  • FISH, fluorescent in situ hybridisation
  • FITC, fluorescein isothiocyanate
  • PBS, phosphate buffered saline
  • RT-PCR, reverse transcriptase polymerase chain reaction
  • SDS, sodium dodecyl sulfate
  • SSC, saline sodium citrate
  • TGF-β1, transforming growth factor β1
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