Purpose: To determine the effect of riboflavin–UV-A treatment on the corneal limbal epithelial cells during a corneal collagen cross-linking (CXL) procedure.
Methods: Thirty freshly enucleated human cadaveric eyeballs were subjected to a CXL procedure, mimicking the clinical protocol. During the UV-A exposure, one half of the limbus (sector A) was left unprotected, whereas the other half (sector B) was covered by a metal shield. Limbal biopsies from both sectors before and after the procedure were analyzed. Each strip of tissue was divided into 3 segments, for cell count of viable cells, for cultivation on human amniotic membrane (HAM), and for stem cell and differentiated corneal epithelial cell marker studies using reverse transcriptase–polymerase chain reaction.
Results: Compared with the cell count before CXL, there was a statistically significant drop in the mean number of viable cells after CXL in sector A but not in sector B. Biopsies from both sectors before CXL and from sector B after CXL showed good growth on HAM. Biopsies from sector A after CXL showed no growth on HAM. The putative stem cell marker ABCG2 was absent in all samples and p63 was absent in 3 of 10 samples taken from sector A after CXL. All markers were present in all samples from sector B after CXL.
Conclusions: Riboflavin–UV-A treatment can result in damage to limbal epithelial cells, particularly the stem cells. Covering the limbal region with a metal shield effectively prevents this damage.
*L and T Microbiology Research Centre, Vision Research Foundation
†Department of Cornea and Refractive Surgery, Medical Research Foundation, Sankara Nethralaya, Chennai, India.
Reprints: Prema Padmanabhan, Department of Cornea and Refractive Surgery, Medical Research Foundation, Sankara Nethralaya, 41 College Rd, Chennai 600 006, India (e-mail:firstname.lastname@example.org).
Supported by the Department of Science and Technology (DST; 105-2008-P).
The authors state that they have no proprietary interest in the products named in this article.
Received October 18, 2011
Accepted January 3, 2012