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Please use this identifier to cite or link to this item: https://dspace.lboro.ac.uk/2134/26833

Title: Chemical and topographical effects on cell differentiation and matrix elasticity in a corneal stromal layer model
Authors: Wilson, Samantha L.
Wimpenny, Ian
Ahearne, Mark
Rauz, Saaeha
El Haj, Alicia J.
Yang, Ying
Issue Date: 2012
Publisher: © Wiley-VCH
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.
Abstract: 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.
Sponsor: 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.
Version: Published
DOI: 10.1002/adfm.201200655
URI: https://dspace.lboro.ac.uk/2134/26833
Publisher Link: https://doi.org/10.1002/adfm.201200655
ISSN: 1616-301X
Appears in Collections:Closed Access (Mechanical, Electrical and Manufacturing Engineering)

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