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Alessandro Ruzza, Mohit Nishitkumar Parekh, Hossein Elbadawy, Vito Romano, Stefano Ferrari, Gianni Salvalaio, Massimo Busin, Diego Ponzin; Pre-loaded tissue for DMEK with endothelium flapped-in . Invest. Ophthalmol. Vis. Sci. 2016;57(12):1215.
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© ARVO (1962-2015); The Authors (2016-present)
To investigate the feasibility of pre-loading the tissues for DMEK.
Tissues were collected from i) tissue culture medium (TCM) and ii) transport medium(TM) [TCM+6% Dextran], stripped and preserved with endothelium flap-in for 4 days at RT. The tissues were collected from TCM and preserved in (a) TCM (n=10) and (b) TM (n=10) respectively; and (c) from TM to TM (n=10) in a 2.2 IOL cartridge. Endothelial cell loss (ECL) was determined using Trypan blue (n=30); metabolism was studied using D-Glucose HK kit (n=30); PAS staining was used for histology (n=9); ZO-1 marker for tight junction (n=12) and TdT dNTP kit for cell apoptosis (n=9) post preservation.
76% cases showed successful stripping of DMEK from a and b, but 100% from c. ECL was 45.05% in combination a, 34.77% in b and 10.77% in c. Glucose uptake (mg/mL) was similar for all the groups. PAS staining showed presence of pure DM and endothelium only in c group. ZO-1 was expressed and mild apoptosis was observed in all the samples. Average stripping and loading time was 19 minutes and 5 minutes respectively.
DMEK tissues can be preserved and pre-loaded using combination c with endothelium flapped-in. It can be used directly in the surgery using a pull-through as well as injection technique. This will reduce surgical time, tissue wastage, costs and provide a validated tissue for transplantation.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.
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