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Application of retina acid improves form and function of tissue engineered corneal construct

Retinoic acid has recently been shown to control the phenotype and extracellular matrix composition of corneal stromal cells cultured in vitro as monolayers. This study set out to investigate the effects of retinoic acid on human corneal keratocytes within a 3D environment. Human corneal keratocytes...

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Main Authors: Zainal Abidin, Fadhilah, Gouveia, Ricardo M, Connon, Che J
Format: Article
Language:English
English
English
Published: Taylor and Francis 2015
Subjects:
RE Ophthalmology
Online Access:http://irep.iium.edu.my/67141/1/Application%20of%20retinoic%20acid%20improves%20form%20and%20function%20of%20tissue%20engineered%20corneal%20construct.pdf
http://irep.iium.edu.my/67141/7/Scopus%20-%20Application%20of%20retinoic%20acid.pdf
http://irep.iium.edu.my/67141/8/WOS%20-%20Application%20of%20retinoic%20acid.pdf
http://irep.iium.edu.my/67141/
https://www.tandfonline.com/doi/full/10.1080/15476278.2015.1093267
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spelling my.iium.irep.671412018-12-14T03:14:32Z http://irep.iium.edu.my/67141/ Application of retina acid improves form and function of tissue engineered corneal construct Zainal Abidin, Fadhilah Gouveia, Ricardo M Connon, Che J RE Ophthalmology Retinoic acid has recently been shown to control the phenotype and extracellular matrix composition of corneal stromal cells cultured in vitro as monolayers. This study set out to investigate the effects of retinoic acid on human corneal keratocytes within a 3D environment. Human corneal keratocytes were encapsulated in collagen gels, which were subsequently compressed under load, and cultured in serum-free media supplemented with 10 mM retinoic acid or DMSO vehicle for 30 days. Cell proliferation was quantified on selected days, while the expression of several important keratocytes markers was evaluated at day 30 using RT-PCR and immunoblotting. The weight and size of the collagen constructs were measured before and after hydration and contraction analyses. Retinoic acid enhanced keratocyte proliferation until day 30, whereas cells in control culture conditions showed reduced numbers after day 21. Both gene and protein expressions of keratocyte-characteristic proteoglycans (keratocan, lumican and decorin), corneal crystallins and collagen type I and V were significantly increased following retinoic acid supplementation. Retinoic acid also significantly reduced the expression of matrix metalloproteases 1, 3 and 9 while not increasing a-smooth muscle actin and fibronectin expression. Furthermore, these effects were also correlated with the ability of retinoic acid to significantly inhibit the contractility of keratocytes while allowing the build-up of corneal stromal extracellular matrix within the 3D constructs. Thus, retinoic acid supplementation represents a promising strategy to improve the phenotype of 3D-cultured keratocytes, and their usefulness as a model of corneal stroma for corneal biology and regenerative medicine applications. Taylor and Francis 2015-10-26 Article PeerReviewed application/pdf en http://irep.iium.edu.my/67141/1/Application%20of%20retinoic%20acid%20improves%20form%20and%20function%20of%20tissue%20engineered%20corneal%20construct.pdf application/pdf en http://irep.iium.edu.my/67141/7/Scopus%20-%20Application%20of%20retinoic%20acid.pdf application/pdf en http://irep.iium.edu.my/67141/8/WOS%20-%20Application%20of%20retinoic%20acid.pdf Zainal Abidin, Fadhilah and Gouveia, Ricardo M and Connon, Che J (2015) Application of retina acid improves form and function of tissue engineered corneal construct. Organogenesis, 11 (3). pp. 122-136. ISSN 1547-6278 https://www.tandfonline.com/doi/full/10.1080/15476278.2015.1093267 10.1080/15476278.2015.1093267
institution Universiti Islam Antarabangsa Malaysia
building IIUM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider International Islamic University Malaysia
content_source IIUM Repository (IREP)
url_provider http://irep.iium.edu.my/
language English
English
English
topic RE Ophthalmology
spellingShingle RE Ophthalmology
Zainal Abidin, Fadhilah
Gouveia, Ricardo M
Connon, Che J
Application of retina acid improves form and function of tissue engineered corneal construct
description Retinoic acid has recently been shown to control the phenotype and extracellular matrix composition of corneal stromal cells cultured in vitro as monolayers. This study set out to investigate the effects of retinoic acid on human corneal keratocytes within a 3D environment. Human corneal keratocytes were encapsulated in collagen gels, which were subsequently compressed under load, and cultured in serum-free media supplemented with 10 mM retinoic acid or DMSO vehicle for 30 days. Cell proliferation was quantified on selected days, while the expression of several important keratocytes markers was evaluated at day 30 using RT-PCR and immunoblotting. The weight and size of the collagen constructs were measured before and after hydration and contraction analyses. Retinoic acid enhanced keratocyte proliferation until day 30, whereas cells in control culture conditions showed reduced numbers after day 21. Both gene and protein expressions of keratocyte-characteristic proteoglycans (keratocan, lumican and decorin), corneal crystallins and collagen type I and V were significantly increased following retinoic acid supplementation. Retinoic acid also significantly reduced the expression of matrix metalloproteases 1, 3 and 9 while not increasing a-smooth muscle actin and fibronectin expression. Furthermore, these effects were also correlated with the ability of retinoic acid to significantly inhibit the contractility of keratocytes while allowing the build-up of corneal stromal extracellular matrix within the 3D constructs. Thus, retinoic acid supplementation represents a promising strategy to improve the phenotype of 3D-cultured keratocytes, and their usefulness as a model of corneal stroma for corneal biology and regenerative medicine applications.
format Article
author Zainal Abidin, Fadhilah
Gouveia, Ricardo M
Connon, Che J
author_facet Zainal Abidin, Fadhilah
Gouveia, Ricardo M
Connon, Che J
author_sort Zainal Abidin, Fadhilah
title Application of retina acid improves form and function of tissue engineered corneal construct
title_short Application of retina acid improves form and function of tissue engineered corneal construct
title_full Application of retina acid improves form and function of tissue engineered corneal construct
title_fullStr Application of retina acid improves form and function of tissue engineered corneal construct
title_full_unstemmed Application of retina acid improves form and function of tissue engineered corneal construct
title_sort application of retina acid improves form and function of tissue engineered corneal construct
publisher Taylor and Francis
publishDate 2015
url http://irep.iium.edu.my/67141/1/Application%20of%20retinoic%20acid%20improves%20form%20and%20function%20of%20tissue%20engineered%20corneal%20construct.pdf
http://irep.iium.edu.my/67141/7/Scopus%20-%20Application%20of%20retinoic%20acid.pdf
http://irep.iium.edu.my/67141/8/WOS%20-%20Application%20of%20retinoic%20acid.pdf
http://irep.iium.edu.my/67141/
https://www.tandfonline.com/doi/full/10.1080/15476278.2015.1093267
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score 13.211869

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