Development of a Surface-Modified Contact Lens for the Transfer of Cultured Limbal Epithelial Cells to the Cornea for Ocular Surface Diseases

Abstract

Our aim was to develop an improved cell transfer system for delivering laboratory-cultured human limbal epithelial cells to the cornea, which would be low risk for the patient and convenient to use for the surgeon. We took a standard contact lens and developed a plasma polymer layer for coating this for attachment of cells to the lens and subsequent transfer of cells to the cornea. A range of plasma polymer surfaces were examined for initial cell attachment using three different combinations of human and rabbit epithelial and stromal cells, initially expanding cells both with and without bovine serum. The most promising surfaces, based on acrylic acid, were then coated onto contact lenses. Cell transfer from the lenses to the denuded surface of a 3D rabbit organ culture model was then used to make a second selection of substrates, which permitted reliable cell transfer. Primary rabbit and human corneal cells attached and proliferated well on acrylic acid-coated surfaces. Reliable transfer of primary epithelial cells from the coated contact lenses to a rabbit cornea was achieved by coating lenses with acrylic acid at 5 W/10 cm(3)/min and using cell densities of 1 x 10(5)/lens and above.

Plasma Polymer-Coated Contact Lenses for the Culture and Transfer of Corneal Epithelial Cells in the Treatment of Limbal Stem Cell Deficiency

“…Deshpande et al have demonstrated that acid-functionalized surfaces deposited by plasma polymerization are able to support human and rabbit corneal epithelial cell proliferation in vitro. 30 The current study demonstrates the suitability of this technique for successful transfer of cultured corneal epithelial cells onto the wounded corneal surface and confirms that transferred labeled cells survive, proliferate, and undergo stratification in vivo.…”

“…30 However, Deshpande et al seeded suspended corneal epithelial cells that require 3T3 feeder cells, 30 which are less desirable for clinical use. Previous studies have demonstrated feeder-free explant culture of corneal epithelium on commercial siloxane-hydrogel lenses in vitro 25 and a pilot study on humans produced clinical improvement in three patients.…”

“…30 In addition, they share structural similarities with that of humans, including the presence of the Bowman's layer. Therefore, rabbits have been used extensively for studying corneal epithelial cells.…”

“…Therefore, rabbits have been used extensively for studying corneal epithelial cells. 20,30,35,36 In this study, we have generated rabbit LSCD models and used them to assess the transfer of autologous cultured cells from the lens surface to the corneal wound bed.…”

“…28,29 The surface concentration of the acid groups can be controlled by the addition of diluents to the plasma process, which, in turn, can affect how strongly cells adhere to these surfaces, allowing the cells to transfer from the surface onto a wound bed. 30 This study investigates the use of plasma polymer-coated contact lenses as substrates for in vitro culture and also as carriers for in vivo transfer of limbal epithelial cells in the treatment of LSCD. Unlike the established hAM-based techniques, where a contiguous sheet of epithelium supported by a membrane is sutured or glued onto the corneal surface using a fibrin glue, the contact lens-based approach transfers only the cells.…”