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Chemical and topographical effects on cell differentiation and matrix elasticity in a corneal stromal layer model

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posted on 05.10.2017 by Sammy Wilson, Ian Wimpenny, Mark Ahearne, Saaeha Rauz, Alicia J. El Haj, Ying Yang
Control and maintenance of the keratocyte phenotype is vital to developing in vitro tissue engineered strategies for corneal repair. In this study the influence of topographical and chemical cues on the mechanical, phenotypical and genotypical behaviour of adult human derived corneal stromal (AHDCS) cells in three dimensional (3D) multi-layered organised constructs is examined. Topographical cues are provided via multiple aligned electrospun nanofiber meshes, which are arranged orthogonally throughout the constructs and are capable of aligning individual cells and permitting cell migration between the layers. The influence of chemical cues is examined using different supplements in culture media. A non-destructive indentation technique and optical coherence tomography are used to determine the matrix elasiticity (elastic modulus) and dimensional changes, respectively. These measurements were indicative of changes in cell phenotype from contractile fibroblasts to quiescent keratocytes over the duration of the experiment and corroborated by qPCR. Constructs containing nanofibers have a higher initial modulus, reduced contraction and organised cell orientation compared to those without nanofibers. Cell-seeded constructs cultured in serum-containing media increased in modulus throughout the culture period and underwent significantly more contraction than constructs cultured in serum-free and insulin-containing media. This implies that the growth factors present in serum promote a fibroblast-like phenotype; qPCR data further validates these observations. These results indicate that the synergistic effect of nanofibers and serum-free media plus insulin supplementation provide the most suitable topographical and chemical environment for reverting corneal fibroblasts to a keratocyte phenotype in a 3D construct.

Funding

Funding from the Doctoral Training Centre of Regenerative Medicine, (EPSRC, UK), BBSRC, UK (BB/F002866/1) and the Birmingham Eye Foundation (UK) (Charity 257549) are gratefully acknowledged.

History

School

  • Mechanical, Electrical and Manufacturing Engineering

Published in

Advanced Functional Materials

Volume

22

Issue

17

Pages

3641 - 3649

Citation

WILSON, S. ... et al., 2012. Chemical and topographical effects on cell differentiation and matrix elasticity in a corneal stromal layer model. Advanced Functional Materials, 22 (17), pp.3641-3649.

Publisher

© Wiley-VCH

Version

AM (Accepted Manuscript)

Publisher statement

This work is made available according to the conditions of the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0) licence. Full details of this licence are available at: https://creativecommons.org/licenses/by-nc-nd/4.0/

Publication date

2012

Notes

This is the peer reviewed version of the following article: WILSON, S. ... et al., 2012. Chemical and topographical effects on cell differentiation and matrix elasticity in a corneal stromal layer model. Advanced Functional Materials, 22 (17), pp.3641-3649, which has been published in final form at: https://doi.org/10.1002/adfm.201200655. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.

ISSN

1616-301X

eISSN

1616-3028

Language

en

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