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K.A. Diehl, J.D. Foley, G. Zhang, P. Podsiadlo, P.F. Nealey, C.J. Murphy; Nanoscale Topography Modulates Corneal Epithelial Cell Migration . Invest. Ophthalmol. Vis. Sci. 2003;44(13):3276.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: To engineer surfaces with nanoscale topographic features that mimic the corneal epithelial basement membrane and to study the effect of these features on corneal epithelial cell migration. Methods: Surface fabrication: Using electron-beam lithography and reaction ion etching, silicon wafers were patterned and etched with grooves and ridges of nano- and microscale dimensions (pitch range from 400 to 4000nm) and coated with silicon oxide to achieve a chemically uniform surface. Additionally, polyurethane patterned surfaces were created through replication molding techniques. Migration of individual cells: SV-40 transformed human corneal epithelial cells in SHEM containing 10% FBS were sparsely seeded onto patterned and smooth control surfaces and allowed to adhere for 12-20 hours. Sequential photographs of cells on surfaces with features of differing dimensions, as well as smooth control surfaces, were obtained every 30 minutes over 24 hours to record migration of individual cells. Cell Dispersion Assay: SV-40 cells in SHEM containing 10% FBS and pretreated with Mitomycin-C to prevent proliferation, were seeded onto patterned and smooth control surfaces in constrained, 500µm diameter, circular areas, using a custom-fabricated cell manifold. Once cells were adherent, the manifold was removed and outward migration of cells monitored, recording the cell colony shape and dimensions daily for 7 days. Results: Individual cells migrated preferentially parallel to grooves and ridges of nano- and microscale dimensions of all pitches assayed. Direction of migration of individual cells on smooth surfaces was random. Groups of cells migrated out from the initial circular seeded zone predominantly along grooves and ridges, elongating to elliptical colonies paralleling surface grooves and ridges of differing dimensions. On smooth surfaces, groups of cells migrated radially, equally in all directions, maintaining a circular colony shape. Conclusions: Biologic length scale substratum features resembling the basement membrane modulate corneal epithelial cell migration. These findings have relevance to the maintenance of corneal homeostasis and wound healing, and the conduction of in vitro studies of cell migration, as well as to tissue engineering and the development of corneal prostheses.
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