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The most recent version of this article was published on 1 April 2007

Br J Ophthalmol. Published Online First: 20 September 2006. doi:10.1136/bjo.2006.103895
Copyright © 2006 by the BMJ Publishing Group Ltd.
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Nick Di Girolamo
Jeanie Chui
Denis Wakefield
Minas T. Coroneo
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Laboratory science - Extended reports

Cultured human ocular surface epithelium on therapeutic contact lenses

Nick Di Girolamo 1*, Jeanie Chui 1, Denis Wakefield 1 and Minas T. Coroneo 2

1 University of New South Wales, Australia
2 UNSW Prince of Wales Hospital, Australia

* To whom correspondence should be addressed. E-mail: n.digirolamo{at}unsw.edu.au.

Accepted 1 September 2006


*  Abstract

Aims: This study was initiated after observing some intriguing epithelial growth properties of contact lens (CLs) used as a bandage for patients post-pterygium surgery. The aim of this investigation was to determine the efficacy of culturing human ocular surface epithelial cells on therapeutic CLs in autologous serum with a view of utilizing this system to transfer epithelial cells to patients with persistent corneal/limbal defects.

Methods: Excess graft tissue resected from patients undergoing pterygium surgery (n=3) consisting of limbal epithelium was placed on siloxane-hydrogel CLs (lotrafilcon A and balafilcon A). Limbal explants were cultured in media with 10% autologous serum. Morphology, proliferative capacity, and a cytokeratin profile were determined by phase contrast, light and electron microscopy (EM) and immunohistochemical analysis.

Results: Lotrafilcon A CLs sustained proliferation and migration from limbal tissue. Cells became confluent after 10-14 days and consisted of 2-3 layers with a corneal phenotype (CK3+/CK12+/CK19-) and a propensity to proliferate (p63+). EM revealed microvilli on the apical surface with adhesive projections, indicating that these cells were stable and likely to survive long-term. Growth was not observed from limbal explants cultured on balafilcon A CLs.

Conclusion: We have developed a method for culturing human ocular surface epithelium on CLs that may facilitate expansion and transfer of autologous limbal epithelial cells whilst avoiding the risks associated with transplanting allogeneic tissue. This technique may be potentially useful for the treatment of patients with limbal stem cell deficiency.

Keywords: conjunctiva, limbus, ocular surface, pterygium




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