A Surgical Technique to Harvest Descemet Membrane with Viable Endothelial Cells for Selective Transplantation

Abstract

Abstract: : Purpose: To describe a surgical technique using an artificial anterior chamber to harvest the Descemet membrane (DM) along with its healthy endothelium for selective DM + endothelial cell transplantation. Methods: Corneoscleral buttons of seven human cadaver eyes were mounted endothelial side up on an artificial anterior chamber (ALTK System, Moria, Antony, France). A superficial trephination was made in the posterior stroma with a 9–0 trephine, inside of the Schwalbe’s line. The artificial anterior chamber was then pressurized by means of saline solution infusion, causing the endothelial side of the donor cornea to bulge and take a convex configuration. The DM then is separated on the trephination side, where a cyclodialysis spatula was used to severe the DM along with its endothelium from the stroma. On six DM, the viability of endothelial cells using this harvesting technique was assessed with vital stains using 0.3% solution of trypan blue for one minute followed by 0.2% alizarin red S for 1½ minutes. The six stained DM were then examined under a light microscope and photographed in order to calculate the endothelial cell damage. The seventh DM was examined with a scanning electron microscope (SEM). Results: The DM carrying donor endothelium was completely stripped from the posterior stroma in all seven eyes. The detached membrane is very fragile and prone to tear, especially after being severed from posterior stroma. A very delicate manipulation is necessary to avoid damage to endothelial cells. Light microscopy showed minimal endothelial cell damage after cell staining with this harvesting method. SEM showed the preservation of endothelial cell morphology after the procedure. Conclusions: The described technique appears to be a safe and straightforward method of harvesting the DM for its selective transplantation. Further studies are underway to determine different approaches for the insertion of the obtained healthy DM through small corneal incisions for corneal endothelial diseases.

Keywords: cornea: endothelium • cornea: clinical science