Effect of Decorin and Aligned Collagen Fibril Topography on TGF-β1 Activation of Corneal Keratocytes

During corneal wound healing, transforming growth factor-beta 1 (TGF-β1) causes differentiation of quiescent keratocytes into myofibroblasts. Decorin has been investigated as a promising anti-fibrotic therapeutic for corneal healing due to its interaction with TGF-β1, collagen, and cell surface rece...

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Main Authors: Nathaniel S. Tjahjono, Divya Subramanian, Tarik Z. Shihabeddin, Hudson D. Hicks, Victor D. Varner, W. Matthew Petroll, David W. Schmidtke
Format: Article
Language:English
Published: MDPI AG 2025-03-01
Series:Bioengineering
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Online Access:https://www.mdpi.com/2306-5354/12/3/259
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author Nathaniel S. Tjahjono
Divya Subramanian
Tarik Z. Shihabeddin
Hudson D. Hicks
Victor D. Varner
W. Matthew Petroll
David W. Schmidtke
author_facet Nathaniel S. Tjahjono
Divya Subramanian
Tarik Z. Shihabeddin
Hudson D. Hicks
Victor D. Varner
W. Matthew Petroll
David W. Schmidtke
author_sort Nathaniel S. Tjahjono
collection DOAJ
description During corneal wound healing, transforming growth factor-beta 1 (TGF-β1) causes differentiation of quiescent keratocytes into myofibroblasts. Decorin has been investigated as a promising anti-fibrotic therapeutic for corneal healing due to its interaction with TGF-β1, collagen, and cell surface receptors. In this study, a novel microfluidic method for coating aligned collagen fibrils with decorin was developed to mimic the presence of decorin within the corneal stroma. Decorin was found to bind selectively to collagen and remained bound for at least five days. To investigate the effects of decorin coatings on keratocyte activation, primary rabbit keratocytes were cultured in the presence of TGF-β1 for 5 days on substrates with or without decorin and stained for α-smooth muscle actin (α-SMA). The expression of α-SMA was reduced by similar amounts on monomeric collagen (40%), random collagen fibrils (32%), and aligned collagen fibrils (32%) coated with decorin as controls. However, α-SMA expression was differentially expressed between the collagen substrates not coated with decorin, with significantly lower expression on uncoated aligned collagen fibrils compared to uncoated collagen monomers. Addition of decorin directly to culture media, had a limited effect on reducing myofibroblast differentiation. Taken together, these results demonstrate the importance of topography and ECM composition on keratocyte activation.
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spelling doaj-art-be25c62f710641acacfa14589bcb2b8d2025-08-20T02:11:00ZengMDPI AGBioengineering2306-53542025-03-0112325910.3390/bioengineering12030259Effect of Decorin and Aligned Collagen Fibril Topography on TGF-β1 Activation of Corneal KeratocytesNathaniel S. Tjahjono0Divya Subramanian1Tarik Z. Shihabeddin2Hudson D. Hicks3Victor D. Varner4W. Matthew Petroll5David W. Schmidtke6Department of Bioengineering, University of Texas at Dallas, Richardson, TX 75080, USADepartment of Bioengineering, University of Texas at Dallas, Richardson, TX 75080, USADepartment of Bioengineering, University of Texas at Dallas, Richardson, TX 75080, USADepartment of Bioengineering, University of Texas at Dallas, Richardson, TX 75080, USADepartment of Bioengineering, University of Texas at Dallas, Richardson, TX 75080, USADepartment of Biomedical Engineering, University of Texas Southwestern Medical Center, Dallas, TX 75390, USADepartment of Bioengineering, University of Texas at Dallas, Richardson, TX 75080, USADuring corneal wound healing, transforming growth factor-beta 1 (TGF-β1) causes differentiation of quiescent keratocytes into myofibroblasts. Decorin has been investigated as a promising anti-fibrotic therapeutic for corneal healing due to its interaction with TGF-β1, collagen, and cell surface receptors. In this study, a novel microfluidic method for coating aligned collagen fibrils with decorin was developed to mimic the presence of decorin within the corneal stroma. Decorin was found to bind selectively to collagen and remained bound for at least five days. To investigate the effects of decorin coatings on keratocyte activation, primary rabbit keratocytes were cultured in the presence of TGF-β1 for 5 days on substrates with or without decorin and stained for α-smooth muscle actin (α-SMA). The expression of α-SMA was reduced by similar amounts on monomeric collagen (40%), random collagen fibrils (32%), and aligned collagen fibrils (32%) coated with decorin as controls. However, α-SMA expression was differentially expressed between the collagen substrates not coated with decorin, with significantly lower expression on uncoated aligned collagen fibrils compared to uncoated collagen monomers. Addition of decorin directly to culture media, had a limited effect on reducing myofibroblast differentiation. Taken together, these results demonstrate the importance of topography and ECM composition on keratocyte activation.https://www.mdpi.com/2306-5354/12/3/259decorinaligned collagen fibrilscorneal keratocytestransforming growth factor-betaalpha-smooth muscle actin
spellingShingle Nathaniel S. Tjahjono
Divya Subramanian
Tarik Z. Shihabeddin
Hudson D. Hicks
Victor D. Varner
W. Matthew Petroll
David W. Schmidtke
Effect of Decorin and Aligned Collagen Fibril Topography on TGF-β1 Activation of Corneal Keratocytes
Bioengineering
decorin
aligned collagen fibrils
corneal keratocytes
transforming growth factor-beta
alpha-smooth muscle actin
title Effect of Decorin and Aligned Collagen Fibril Topography on TGF-β1 Activation of Corneal Keratocytes
title_full Effect of Decorin and Aligned Collagen Fibril Topography on TGF-β1 Activation of Corneal Keratocytes
title_fullStr Effect of Decorin and Aligned Collagen Fibril Topography on TGF-β1 Activation of Corneal Keratocytes
title_full_unstemmed Effect of Decorin and Aligned Collagen Fibril Topography on TGF-β1 Activation of Corneal Keratocytes
title_short Effect of Decorin and Aligned Collagen Fibril Topography on TGF-β1 Activation of Corneal Keratocytes
title_sort effect of decorin and aligned collagen fibril topography on tgf β1 activation of corneal keratocytes
topic decorin
aligned collagen fibrils
corneal keratocytes
transforming growth factor-beta
alpha-smooth muscle actin
url https://www.mdpi.com/2306-5354/12/3/259
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