April 1998
Volume 39, Issue 5
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Articles  |   April 1998
Transforming growth factor-beta 1 promotes contraction of collagen gel by bovine corneal fibroblasts through differentiation of myofibroblasts.
Author Affiliations
  • H Kurosaka
    Department of Ophthalmology, Keio University, School of Medicine, Tokyo, Japan.
  • D Kurosaka
    Department of Ophthalmology, Keio University, School of Medicine, Tokyo, Japan.
  • K Kato
    Department of Ophthalmology, Keio University, School of Medicine, Tokyo, Japan.
  • Y Mashima
    Department of Ophthalmology, Keio University, School of Medicine, Tokyo, Japan.
  • Y Tanaka
    Department of Ophthalmology, Keio University, School of Medicine, Tokyo, Japan.
Investigative Ophthalmology & Visual Science April 1998, Vol.39, 699-704. doi:https://doi.org/
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      H Kurosaka, D Kurosaka, K Kato, Y Mashima, Y Tanaka; Transforming growth factor-beta 1 promotes contraction of collagen gel by bovine corneal fibroblasts through differentiation of myofibroblasts.. Invest. Ophthalmol. Vis. Sci. 1998;39(5):699-704. doi: https://doi.org/.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

PURPOSE: To determine whether the ability of transforming growth factor-beta (TGF-beta) to influence the contractile activity of corneal fibroblasts depends on their differentiation into myofibroblasts. METHODS: Bovine corneal fibroblasts were cultured on collagen gel in MED 5 medium (F-12 nutrient mixture supplemented with 5% fetal bovine serum) with or without TGF-beta 1 (0.01-10 ng/ml). To evaluate the corneal fibroblast-derived contraction of collagen gel, the thickness of the gel was measured daily for 6 days. The total number of cells on the gel was counted with a Coulter counter. The detection of alpha-smooth muscle actin (alpha-SMA); a marker for myofibroblasts, on these cells was performed immunocytochemically by using a mouse monoclonal antibody against alpha-SMA. The number of myofibroblasts (alpha-SMA-positive cells) was determined. RESULTS: The control gels containing bovine corneal fibroblasts that were cultured with the MED 5 medium alone significantly contracted to 72.3 +/- 1.2% of their original thickness after 6 days. TGF-beta 1 increased the contraction of collagen gel mediated by bovine corneal fibroblasts in a dose-dependent manner. Approximately 0.2% of the cells on the control gels cultured with MED 5 medium alone were alpha-SMA positive. TGF-beta 1 significantly increased the expression of alpha-SMA in a dose-dependent manner. There was no significant correlation between the thickness of the collagen gel and the total number of cells. However, there was a significant negative correlation between the thickness of collagen gel and the number of myofibroblasts. CONCLUSIONS: TGF-beta 1 increased the contractile activity of bovine corneal fibroblasts and their ability to differentiate into myofibroblasts. Because contractile activity was correlated with differentiation, the influence of TGF-beta 1 on corneal fibroblast-induced collagen gel contraction may depend on the promotion of myofibroblast differentiation.

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