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

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 E-ISSN 1555-8592

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Abstract

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.

Item Type:	Article (Journal)
Additional Information:	5982/67141
Uncontrolled Keywords:	3D model, collagen gel, Cornea, in vitro, keratocytes, retinoic acid, tissue engineering
Subjects:	R Medicine > RE Ophthalmology
Kulliyyahs/Centres/Divisions/Institutes (Can select more than one option. Press CONTROL button):	Kulliyyah of Pharmacy Kulliyyah of Pharmacy > Department of Basic Medical Sciences
Depositing User:	Dr Fadhilah Zainal Abidin
Date Deposited:	14 Dec 2018 11:02
Last Modified:	14 Dec 2018 11:14
URI:	http://irep.iium.edu.my/id/eprint/67141

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