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Jingbo Liu, Brian D. Lawrence, Aihong Liu, Mark I. Rosenblatt; Silk Fibroin As A Biomaterial Substrate For Corneal Epithelial Cell Sheet Generation. Invest. Ophthalmol. Vis. Sci. 2012;53(14):267.
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© ARVO (1962-2015); The Authors (2016-present)
To evaluate a silk fibroin (SF) biomaterial as a substrate for corneal epithelial cell proliferation, differentiation and stratification in vitro compared with denuded amniotic membrane (AM).
Primary human and rabbit corneal epithelial cells and immortalized human corneal limbal epithelial cells were cultured on the SF and denuded AM, respectively. The biological cell behavior including the morphology, proliferation, differentiation and stratification on the two substrates were compared and analyzed.
Corneal epithelial cells can adhere and proliferate on the SF and denuded AM with a cobblestone appearance, abundant microvilli on the surface and wide connection with the adjacent cells. MTT assay showed that cell proliferation on denuded AM was statistically higher than that on SF at 24 and 72 hours post plating (p = 0.001 and 0.0005, respectively). Expression of ΔNp63a and keratin 3/12 were detected in primary cells cultures on the two substrates with no statistical difference. When cultured at the air-liquid interface for 10 days, cells on SF could form a comparable stratified graft with a 2 to 3 cell layering which compared similarly to AM cultures.
SF, a novel biomaterial, could support corneal epithelial cells to proliferate, differentiate and stratify retaining the normal characteristic epithelium phenotype. Compared with AM, its unique features including the transparency, ease of handling and transfer, and inherent freedom from disease transmission make it a promising substrate for corneal wound repair and tissue engineering purposes.
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