Effect of intraoperative trypan blue on lens epithelial cells – Histomorphological analysis

Abstract

IntroductionTrypan Blue is an acid azo dye commonly used as a stain to distinguish viable from non-viable cells. It is a vital stain used intra operatively during cataract surgery to stain the external surface of the anterior lens capsule for better visualization. AimTo analyze the histomorphological effects of trypan blue on Lens Epithelial cells and the Basement Membrane on direct exposure by staining the internal surface of the anterior lens capsule during Small Incision Cataract Surgery. MethodsAnalytical cross sectional case control study. Anterior capsule specimens of 14 Patients undergoing small incision cataract surgery at Department of Ophthalmology, Govt Medical College Hospital, Thrissur were studied. Two specimens of anterior capsule taken from the same eye form the case and control. Control specimen (sample A) was removed first, after the routine external staining with trypan blue 0.06% (w/v) for 10 seconds. The stain was washed off by balanced salt solution in every case. Then trypan blue was injected under the remaining anterior capsule and case (Test) specimen (sample B) was obtained after direct contact of trypan blue to the internal surface (lens epithelial cells) for 1 minute. Histomorphological (qualitative and quantitative) examination of both specimens done. ResultsQualitative data analysis was done by EPI INFO software.v.7. Intactness of LECs throughout the length was statistically significant in Sample A (p = 0.000027). Partial and complete detachment of Lens Epithelial Cells, degeneration, and nuclear smudging were significantly higher in Sample B. Qualitative analysis of the basement membrane showed significant edema of the basement membrane in sample B. Basement membrane splitting observed in sample B was not statistically significant. Quantitative data analyzed using independent t test. There was a statistically significant decrease in cell density in sample B with p value less than 0.05. DiscussionOur study demonstrated that direct staining of the internal surface of anterior capsule with trypan blue affected LECs and the basement membrane. There were reduction in cell density, irreversible degeneration of Lens Epithelial Cells and basement membrane edema. Hence treating the internal surface of capsular bag with trypan blue may reduce incidence of Posterior capsular opacification.