Effect of trypan blue staining on the density and viability of lens epithelial cells in white cataract.
OBJECTIVE
To assess the effect of anterior capsule staining with trypan blue 0.0125% on the density and viability of the lens epithelial cells (LECs).


METHODS
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).


CONCLUSIONS
Staining the anterior capsule with trypan blue affected the density and viability of LECs.
Nanavaty
Mayank A
MA
Iladevi Cataract & IOL Research Centre, Raghudeep Eye Clinic, Ahmedabad, India.
Johar
Kaid
K
Sivasankaran
Meena A
MA
Vasavada
Abhay R
AR
Praveen
Mamidipudi R
MR
Zetterström
Charlotta
C
D019923:Capsulorhexis; D002386:Cataract / Q000175:diagnosis*; D002452:Cell Count; D002470:Cell Survival / Q000187:drug effects; D004396:Coloring Agents / Q000494:pharmacology*; D004847:Epithelial Cells / Q000187:drug effects* / Q000473:pathology; D005260:Female; D006801:Humans; D007903:Lens Capsule, Crystalline / Q000187:drug effects / Q000473:pathology; D007908:Lens, Crystalline / Q000187:drug effects* / Q000473:pathology; D008297:Male; D008875:Middle Aged; D018918:Phacoemulsification; D011446:Prospective Studies; D013194:Staining and Labeling / Q000379:methods; D014343:Trypan Blue / Q000494:pharmacology*
D004396:Coloring Agents; D014343:Trypan Blue