Endothelial Keratoplasty: The Influence of Insertion Techniques and Incision Size on Donor Endothelial Survival

Purpose: To determine the acute endothelial cell damage from trephination and tissue insertion in endothelial keratoplasty (EK) surgery. The influence of insertion technique (forceps insertion vs “pull-through” insertion) of donor tissue and incision size (3 vs 5 mm length) was assessed. Methods: Forty precut 8.-mm-diameter donor posterior buttons were used in this study. Thirty-five buttons were inserted through a limbal incision of either 3 or 5 mm length into the anterior chamber of cadaver eyes and then removed through an open sky technique without further trauma. Five buttons that were trephined but not inserted served as a control group. Vital dye staining and computer digitized planimetry were used to analyze the tissue and quantify the total damaged area over the entire endothelial surface. Five buttons for each of 7 insertion techniques were used. The 8 tissue groups evaluated were as follows: group 1: control group of trephination only, with no insertion; group 2: forceps with folded tissue through 5-mm incision; group 3: suture pull through of nonfolded tissue through a 5-mm incision; group 4: forceps pull through of Busin glide folded tissue through a 5-mm incision; group 5: forceps with folded tissue through a 3-mm incision; group 6: suture pull through with folded tissue through a 3-mm incision; group 7: suture pull through with nonfolded tissue through a 3-mm incision; and group 8: forceps pull through of Busin glide folded tissue through a 3-mm incision. Results: The control group demonstrated 9% ± 2% peripheral cell damage from simple trephination of the tissue but without insertion. In the 5-mm incision surgeries, forceps insertion (group 2) caused 18% ± 3% loss, suture pull-through insertion (group 3) caused 18% ± 2% loss, and Busin glide pull through (group 4) caused 20% ± 5% loss. There were no significant differences in damage between any of the 5-mm incision group techniques (P > 0.99). In the 3-mm incision surgeries, forceps insertion (group 5) caused a 30% ± 3% loss, pull through with folded tissue (group 6) caused 30% ± 5% loss, pull through with nonfolded tissue (group 7) caused 56% ± 4% loss, and Busin glide pull through (group 8) caused a 28%± 5% loss. There was no difference in damage among the 3-mm groups (P > 0.96), with the exception of group 7 where pulling the unfolded tissue through a 3-mm incision was significantly worse than all other techniques (P < 0.001). There was significantly greater cell area damage in the 3-mm groups (36%) than in the 5-mm groups (19%) (P <0.001). Large patterns of striae with cell loss were seen in the 3-mm groups emanating from the peripheral traction site, regardless of whether the traction to pull the tissue through the incision and into the chamber was generated by a suture or cross-chamber forceps. Direct forceps insertion caused circular patterns of injury at the tip compression site regardless of incision size, but this damage was multiplied and exacerbated by insertion through a smaller incision. Conclusions: Smaller size (3 mm) incisions for EK surgery result in greater acute endothelial area damage than larger size (5 mm) incisions. Pull-through insertion techniques through a 5-mm incision seem equivalent in the amount of induced area damage to that of forceps insertion. Compressive injury from the incision appeared less when the tissue was folded than when not folded. Insertion with any technique through a 3-mm incision resulted in larger areas of endothelial damage. All these iatrogenic death zones outside the central endothelial area would be missed clinically by standard early specular microscopy after EK surgery.