The Use of Trypan Blue to Identify the Area of Antimetabolite Treatment During Trabeculectomy With Mitomycin-C or 5-Fluorouracil

Abstract

Abstract: : Purpose: Trypan blue is currently used to stain the anterior capsule during cataract surgery. We have recently been adding trypan blue to antimetabolites to delineate treatment area during trabeculectomy. The purpose of this study was to determine whether trypan blue alters the cytotoxicity of mitomycin-C (MMC) or 5-fluorouracil (5-FU) on cultured Tenon's capsule fibroblasts. Methods: MMC (0.4mg/ml) and 5FU (25mg/ml) were reconstituted in culture medium with, or without trypan blue at various dilutions. A lactate dehydrogenase release assay was used to measure drug-induced cell death and viable cell number 72 hours post treatment. Results: The addition of trypan blue to MMC or 5FU at concentrations up to 50% did not alter MMC induced cell death (p=0.11; Mann Whitney) or the number of viable fibroblast (5FU, p=0.34: MMC p=0.89; Mann Whitney) Conclusion: The addition of trypan blue to MMC and 5FU does not affect drug cytoxicity for the assays investigated. Trypan blue can be used to estimate treatment area during trabeculectomy. This will allow investigators to further standardise antimetabolite treatment and determine the relationship between treatment area and surgical outcome.

Keywords: wound healing • clinical research methodology • apoptosis/cell death