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Chen, Bo; Jones, Roanne R.; Mi, Shengli; Foster, James; Alcock, Simon; Hamley, Ian; Connon, Che (2012)
Publisher: Royal Society of Chemistry
Languages: English
Types: Article
The human amniotic membrane (AM) is a tissue of fetal origin and has proven to be clinically useful as\ud a biomaterial in the management of various ocular surface disorders including corneal stem cell\ud transplantation. However, its success rate displays a degree of clinical unpredictability. We suggest that\ud the measured variability inAMstiffness offers an explanation for the poor clinical reproducibility when\ud it is used as a substrate for stem cell expansion and transplantation. Corneal epithelial stem cells were\ud expanded upon AM samples possessing different mechanical stiffness. To investigate further the\ud importance of biological substrate stiffness on cell phenotype we replaced AM with type I collagen gels\ud of known stiffness. Substrate stiffness was measured using shear rheometry and surface topography\ud was characterized using scanning electron microscopy and atomic force microscopy. The\ud differentiation status of epithelial cells was examined using RT-PCR, immunohistochemistry and\ud Western blotting. The level of corneal stem cell differentiation was increased in cells expanded upon\ud AM with a high dynamic elastic shear modulus and cell expansion on type I collagen gels confirmed\ud that the level of corneal epithelial stem cell differentiation was related to the substrate’s mechanical\ud properties. In this paper we provide evidence to show that the preparatory method of AM for clinical\ud use can affect its mechanical properties and that these measured differences can influence the level of\ud differentiation within expanded corneal epithelial stem cells.
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