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Adhesion, phenotypic expression, and biosynthetic capacity of corneal keratocytes on surfaces coated with hyaluronic acid of different molecular weights

Paper ID Volume ID Publish Year Pages File Format Full-Text
776 63 2012 12 PDF Available
Title
Adhesion, phenotypic expression, and biosynthetic capacity of corneal keratocytes on surfaces coated with hyaluronic acid of different molecular weights
Abstract

In ophthalmology, hyaluronic acid (HA) is an important extracellular matrix (ECM) component and is appropriate for use in generating a microenvironment for cell cultivation. The aim of this work was to evaluate the rabbit corneal keratocyte (RCK) growth in response to HA coatings under serum-free conditions. After modification with HA of varying molecular weights (MWs: 35–1500 kDa), the surfaces were characterized by atomic force microscopy and contact angle measurements, and were used for cell culture studies. Our data indicated that the substrates coated with higher negatively charged HA become rougher and are more hydrophilic, resulting in the decrease of cell adhesion and cell–matrix interaction. This early cellular event was likely responsible for the determination of keratocyte configuration. Additionally, for the growth of RCKs on dry HA coatings with surface roughness of 1.1–1.7 nm, a strong cell–cell interaction was observed, which may facilitate the formation of multicellular spheroid aggregates and maintenance of mitotically quiescent state. At each culture time point from 1 to 5 days, a better biosynthetic capacity associated with a higher prevalence of elevated ECM production was found for the cells in a spherical configuration. Irrespective of polysaccharide MW of surface coatings, the RCKs presented good viability without hypoxia-induced death. As compared with a monolayer of adherent keratocytes on tissue culture polystyrene plates and low MW HA-modified samples, the cell spheroids (76–110 μm in diameter) showed significantly higher expressions of keratocan and lumican and lower expressions of biglycan, similar to those of keratocytes in vivo. Moreover, the expression levels of corneal crystallin aldehyde dehydrogenase (7–9-fold increase) and nestin (10–16-fold increase) were greater in larger-sized spheroids, indicating higher ability to maintain cellular transparency and self-renewal potential. It is concluded that the cultured RCKs on surfaces coated with HA of different MWs can sense ECM cues, and the multicellular spheroids may potentially be used for corneal stromal tissue engineering applications.

Keywords
Hyaluronic acid; Molecular weight; Extracellular matrix coating; Keratocyte; Corneal stromal tissue engineering
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Adhesion, phenotypic expression, and biosynthetic capacity of corneal keratocytes on surfaces coated with hyaluronic acid of different molecular weights
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Publisher
Database: Elsevier - ScienceDirect
Journal: Acta Biomaterialia - Volume 8, Issue 3, March 2012, Pages 1068–1079
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us