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An In Vitro Model for Assessing Corneal Keratocyte Spreading and Migration on Aligned Fibrillar Collagen

Background: Corneal stromal cells (keratocytes) are responsible for developing and maintaining normal corneal structure and transparency, and for repairing the tissue after injury. Corneal keratocytes reside between highly aligned collagen lamellae in vivo. In addition to growth factors and other so...

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Autores principales: Kivanany, Pouriska B., Grose, Kyle C., Yonet-Tanyeri, Nihan, Manohar, Sujal, Sunkara, Yukta, Lam, Kevin H., Schmidtke, David W., Varner, Victor D., Petroll, W. Matthew
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6306816/
https://www.ncbi.nlm.nih.gov/pubmed/30248890
http://dx.doi.org/10.3390/jfb9040054
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author Kivanany, Pouriska B.
Grose, Kyle C.
Yonet-Tanyeri, Nihan
Manohar, Sujal
Sunkara, Yukta
Lam, Kevin H.
Schmidtke, David W.
Varner, Victor D.
Petroll, W. Matthew
author_facet Kivanany, Pouriska B.
Grose, Kyle C.
Yonet-Tanyeri, Nihan
Manohar, Sujal
Sunkara, Yukta
Lam, Kevin H.
Schmidtke, David W.
Varner, Victor D.
Petroll, W. Matthew
author_sort Kivanany, Pouriska B.
collection PubMed
description Background: Corneal stromal cells (keratocytes) are responsible for developing and maintaining normal corneal structure and transparency, and for repairing the tissue after injury. Corneal keratocytes reside between highly aligned collagen lamellae in vivo. In addition to growth factors and other soluble biochemical factors, feedback from the extracellular matrix (ECM) itself has been shown to modulate corneal keratocyte behavior. Methods: In this study, we fabricate aligned collagen substrates using a microfluidics approach and assess their impact on corneal keratocyte morphology, cytoskeletal organization, and patterning after stimulation with platelet derived growth factor (PDGF) or transforming growth factor beta 1 (TGFβ). We also use time-lapse imaging to visualize the dynamic interactions between cells and fibrillar collagen during wound repopulation following an in vitro freeze injury. Results: Significant co-alignment between keratocytes and aligned collagen fibrils was detected, and the degree of cell/ECM co-alignment further increased in the presence of PDGF or TGFβ. Freeze injury produced an area of cell death without disrupting the collagen. High magnification, time-lapse differential interference contrast (DIC) imaging allowed cell movement and subcellular interactions with the underlying collagen fibrils to be directly visualized. Conclusions: With continued development, this experimental model could be an important tool for accessing how the integration of multiple biophysical and biochemical signals regulate corneal keratocyte differentiation.
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spelling pubmed-63068162019-01-02 An In Vitro Model for Assessing Corneal Keratocyte Spreading and Migration on Aligned Fibrillar Collagen Kivanany, Pouriska B. Grose, Kyle C. Yonet-Tanyeri, Nihan Manohar, Sujal Sunkara, Yukta Lam, Kevin H. Schmidtke, David W. Varner, Victor D. Petroll, W. Matthew J Funct Biomater Article Background: Corneal stromal cells (keratocytes) are responsible for developing and maintaining normal corneal structure and transparency, and for repairing the tissue after injury. Corneal keratocytes reside between highly aligned collagen lamellae in vivo. In addition to growth factors and other soluble biochemical factors, feedback from the extracellular matrix (ECM) itself has been shown to modulate corneal keratocyte behavior. Methods: In this study, we fabricate aligned collagen substrates using a microfluidics approach and assess their impact on corneal keratocyte morphology, cytoskeletal organization, and patterning after stimulation with platelet derived growth factor (PDGF) or transforming growth factor beta 1 (TGFβ). We also use time-lapse imaging to visualize the dynamic interactions between cells and fibrillar collagen during wound repopulation following an in vitro freeze injury. Results: Significant co-alignment between keratocytes and aligned collagen fibrils was detected, and the degree of cell/ECM co-alignment further increased in the presence of PDGF or TGFβ. Freeze injury produced an area of cell death without disrupting the collagen. High magnification, time-lapse differential interference contrast (DIC) imaging allowed cell movement and subcellular interactions with the underlying collagen fibrils to be directly visualized. Conclusions: With continued development, this experimental model could be an important tool for accessing how the integration of multiple biophysical and biochemical signals regulate corneal keratocyte differentiation. MDPI 2018-09-21 /pmc/articles/PMC6306816/ /pubmed/30248890 http://dx.doi.org/10.3390/jfb9040054 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Kivanany, Pouriska B.
Grose, Kyle C.
Yonet-Tanyeri, Nihan
Manohar, Sujal
Sunkara, Yukta
Lam, Kevin H.
Schmidtke, David W.
Varner, Victor D.
Petroll, W. Matthew
An In Vitro Model for Assessing Corneal Keratocyte Spreading and Migration on Aligned Fibrillar Collagen
title An In Vitro Model for Assessing Corneal Keratocyte Spreading and Migration on Aligned Fibrillar Collagen
title_full An In Vitro Model for Assessing Corneal Keratocyte Spreading and Migration on Aligned Fibrillar Collagen
title_fullStr An In Vitro Model for Assessing Corneal Keratocyte Spreading and Migration on Aligned Fibrillar Collagen
title_full_unstemmed An In Vitro Model for Assessing Corneal Keratocyte Spreading and Migration on Aligned Fibrillar Collagen
title_short An In Vitro Model for Assessing Corneal Keratocyte Spreading and Migration on Aligned Fibrillar Collagen
title_sort in vitro model for assessing corneal keratocyte spreading and migration on aligned fibrillar collagen
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6306816/
https://www.ncbi.nlm.nih.gov/pubmed/30248890
http://dx.doi.org/10.3390/jfb9040054
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