Abstract
Many cell types are known to modulate their behaviour in response to changes in material stiffness; however, little is known about how stiffness affects corneal epithelial cells. This study aims to investigate the response of a corneal epithelial cell line to polydimethylsiloxane (PDMS) substrates with a range of Young's moduli from 10 to 1500 kPa. Cellular morphology, proliferation, differentiation and mechanobiology were examined. Cells grown on PDMS adopted the typical cobblestone morphology exhibited by the corneal epithelium. Proliferative markers pERK and Ki67 were higher in cells cultured on stiffer substrates compared with those on softer substrates. Material stiffness was also found to influence the cell phenotype with cells on stiffer substrates having higher cytokeratin 3 gene expression, a mature epithelial marker, while cells on softer substrates expressed more cytokeratin 14, a basal epithelial marker. Cells grown on softer substrates also displayed higher levels of focal adhesions and intermediate filaments compared with cells on stiff substrates. This research will aid in designing novel biomaterials for the culture and transplantation of corneal epithelial cells.
Highlights
Damage to the corneal epithelium can occur due to a variety of conditions including limbal stem cell deficiencies or physical abrasion
PDMS was used to evaluate the effect of substrate stiffness on the behaviour of corneal epithelial cells
To achieve a wider range of stiffness values for this study, two PDMS solutions (Sylgard 184 and 527) were mixed at different ratios. This technique was originally developed by Palchesko et al and has previously been used to examine the effect of substrate stiffness several different cell types [24,25] but not corneal epithelial cells
Summary
Damage to the corneal epithelium can occur due to a variety of conditions including limbal stem cell deficiencies or physical abrasion. Limbal-derived stem cells allow the epithelium to undergo renewal following injury by migrating into the damaged region and differentiating into epithelial cells. Applied directly to a patient to provide a new stem cell source [2]; a lack of suitable donor tissue can 2 limit this option. The alternative is to isolate and culture limbal epithelial stem cells in vitro and to transplant these cells on a biomaterial carrier. This approach has the advantages of allowing a higher number of cells to be transplanted and allowing autologous cells from a patient biopsy to be used. Optimization of the culture environment, including the physical substrate onto which the cells are adhered, is required to control the cell phenotype
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