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Updated: Oct 1, 2026
Corneal Donor Tissue Preparation for Descemet's Membrane Endothelial Keratoplasty
Published on: September 17, 2014
Effect of Proteolytic Enzymes on Descemet Membrane Endothelial Keratoplasty (DMEK) Graft Elasticity
Gil Ben-David1,2,3, Assaf Ben-Arzi1,2,3, Irit Bahar1,2,3
1Department of Ophthalmology, Rabin Medical Center - Beilinson Hospital, Petach Tikva, Israel.
Purpose:
To investigate the proteolytic effect on Descemet membrane endothelial keratoplasty (DMEK) graft elasticity.
Methods:
The ex-vivo experimental study included 19 DMEK-grafts treated with elastase, hyaluronidase, collagenase-IV, or collagenase-VIII at varying concentrations and temperatures. The maximal-scroll-width (MSW) served as a quantitative marker for scroll-tightness. Following enzymatic treatment, endothelial toxicity was evaluated by vital dye staining and endothelial cell shedding into the immersion buffer solution was quantified using a hemocytometer.
Results:
Collagenase-IV (N = 6) at 1-2.5 mg/mL, 37°C for 30-45 min increased MSW by a mean of 22.1% (p < .05), whereas higher concentrations at room-temperature (RT) decreased MSW by 9.7% (p > .05). Collagenase-VIII (N = 2) showed no significant effects. Elastase (N = 7) at 1-2.5 mg/mL, RT for 90-120 min increased MSW by 6.7% (p < .05), while exposure at 37°C reduced MSW by 10.5% (p < .05). Hyaluronidase (N = 4) at 1-2.5 mg/mL, 37°C for 120 min decreased MSW by 19.2% (p < .01), whereas at RT, 1 mg/mL for 60 min increased MSW by 23% (p < .01). 29% of injured endothelial cells were observed across all the groups. Endothelial cell shedding was not significantly different between the two groups (p > .05).
Conclusion:
Collagenase IV showed promising preliminary effects under selected experimental conditions; however, given the small subgroup sizes and variable findings, further controlled studies are required.
