Effect of trypan blue staining on the density and viability of lens epithelial cells in white cataract

PURPOSE: To assess the effect of anterior capsule staining with trypan blue 0.0125% on the density and viability of the lens epithelial cells (LECs). SETTING: Iladevi Cataract and IOL Research Center, Ahmedabad, India. METHODS: This randomized prospective study comprised 40 eyes having phacoemulsification for white mature cataract. Patients were randomized to have the anterior capsule stained with 0.1 mL trypan blue 0.0125% solution before the anterior capsulorhexis (Group 1, n = 20) or to have no anterior capsule staining before the anterior capsulorhexis (Group 2, n = 20). The density and viability of LECs were analyzed. The first 20 samples (10 from Group 1, 10 from Group 2) were stained with hematoxylin-eosin for cell density analysis using image-analysis software. The remaining 20 samples (10 from each group) were stained with a solution containing 0.5 microM calcein AM and 5 microM propidium iodide for 10 minutes at 37 degrees C for viability assay. RESULTS: The mean cell density in Group 1 (with staining) was 3533.15 cells/mm(2) +/- 664.01 (SD). This was significantly lower than the mean of 4235.59 +/- 414.93 cells/mm(2) in Group 2 (no staining) (P = .01). The mean capsule area covered with LECs (80.75% +/- 8.04% versus 94.63% +/- 3.78%) and the mean live LECs in the covered area (51.69% +/- 11.54% versus 68.67% +/- 9.15%) were also significantly lower in Group 1 (P = .002). CONCLUSION: Staining the anterior capsule with trypan blue affected the density and viability of LECs.