Search

Purpose: To evaluate the Descemet stripping automated endothelial keratoplasty (DSAEK) graft deturgescence in preservation medium after microkeratome cut using Fourier domain optical coherence tomography., Methods: The central and peripheral thickness of DSAEK grafts was measured by Fourier domain optical coherence tomography immediately after microkeratome cuts and 1, 2, 3, and 4 hours afterward. All measurements were taken when the grafts were stored in 4°C preservation medium. The hourly change in central graft thickness and graft shape (peripheral graft thicknes - central graft thickness) was calculated and tracked over time., Results: Five DSAEK grafts were measured. The average central graft thickness was 188.7 ± 44.4 μm (range, 146-255 µm) immediately after microkeratome cuts. The average central graft thickness was 147.5 ± 33.0 μm (range, 116-190 µm) after 4 hours in preservation medium (P < 0.001). The average hourly change in central graft thickness was -30.5 μm (P = 0.0051), -8.6 μm (P = 0.055), -2.0 μm (P = 0.42), and 0.0 μm (P = 0.93) at 1, 2, 3, and 4 hours, respectively, after microkeratome cuts. The average hourly change in graft shape was insignificant., Conclusions: DSAEK grafts become thinner after microkeratome cut and stabilize at approximately 2 hours. Therefore, DSAEK graft thickness should be measured at 1.5 to 3 hours after microkeratome cut.

Purpose: To evaluate the endothelial cell survival and stromal bed quality when creating deep stromal cuts with a low-pulse energy, high-frequency femtosecond laser to produce "ultrathin" tissue for Descemet stripping automated endothelial keratoplasty., Methods: Seventeen corneas were used for this study. Five corneas were cut with the laser at a depth of 420 to 500 μm to produce a tissue thickness of approximately ≤70 μm. Five corneas served as an uncut comparison group. Vital dye staining and computer digitized planimetry analysis were performed on these corneas. The 7 remaining corneas were cut for scanning electron microscopy evaluation., Results: The mean central posterior stromal thickness of cut corneas was 60.6 μm (range, 43-72 μm). Endothelial cell damage in cut and comparison corneas was 3.92% ± 2.22% (range, 1.71%-6.51%) and 4.15% ± 2.64% (range, 1.21%-7.01%), respectively (P = 0.887). Low-magnification (×12) scanning electron microscopy revealed a somewhat irregular-appearing surface with concentric rings peripherally. Qualitative grading of higher magnification (×50) central images resulted in an average score of 2.56 (between smooth and rough)., Conclusions: Ultrathin tissue for Descemet stripping automated endothelial keratoplasty can be safely prepared with minimal endothelial cell damage using a low-pulse energy, high-frequency femtosecond laser; however, the resulting stromal surface quality may not be optimal with this technique.

Purpose: To determine the endothelium damage resulting from the application of "dry" gentian violet (GV) stromal markings on corneas precut for Descemet stripping automated endothelial keratoplasty (DSAEK) using a Moria "S" Stamp., Methods: Five precut corneas had the stromal graft bed marked with GV using the Moria "S" stamp and 6 precut corneas were left unmarked as controls. After applying the ink to the stamp, care was taken to allow the alcohol carrier to dry for 10 seconds before applying the dry dye to the stromal surface. Tissue was then trephinated and stained with calcein AM to assess endothelial viability. Grafts were photographed and digital pixel planometry, using an established analysis technique, was used to compare the damage between the control and experimental groups., Results: The mean percent cell damage of corneas treated with GV "S" stamp (n = 5) was 8.6% (range 4.4-12.9), and it was 8.1% (range 3.9-15.1) in the DSAEK control set (n = 6). Median percent cell damage was 6.7% among GV-treated corneas and 7.4% among control corneas. The distributions were not significantly different between groups (Mann-Whitney U test = 15.0, two-tailed P = 1.0). Moreover, no "S" pattern of damage was seen in any study eye., Conclusions: There were no significant differences in endothelial damage between the 2 groups. GV stromal markings may be applied without undue damage to the endothelium using the dry-ink technique described.

Purpose: To use excimer laser smoothing passes to reshape Descemet-stripping automated endothelial keratoplasty (DSAEK) endothelial grafts and to evaluate the effect on the donor endothelium., Methods: The stromal surface of microkeratome-cut DSAEK grafts was smoothed using excimer laser smoothing passes with masking fluid. Excimer laser hyperopic ablation was used to improve the uniformity of graft thickness within the optical zone. Fourier-domain optical coherence tomography was used to measure endothelial graft pachymetry, plan ablations, and evaluate donor contour. Vital dye staining was performed to assess endothelial cell damage. Scanning electron microscopy images of stromal surfaces were graded on a 5-point scale by masked observers to evaluate surface roughness., Results: Four grafts underwent excimer laser smoothing. Vital dye staining showed no endothelial damage. Microkeratome-cut surfaces treated with laser smoothing (mean grade = 2.04) were smoother than nonsmoothed microkeratome-cut surfaces (mean grade = 4.07; P < 0.01), surfaces that underwent dry laser ablation (mean grade = 3.63; P < 0.01) and manually dissected interfaces (mean grade = 4.75; P < 0.0001). No difference was observed between stromal beds created by peeling Descemet membrane (mean grade = 1.64) compared with surfaces produced by microkeratome cutting followed by laser smoothing (mean grade = 2.04; P = 0.14). One graft underwent combined excimer smoothing and peripheral hyperopic ablation. The center-periphery thickness difference was 15 μm before ablation and 4 μm afterward., Conclusions: Laser smoothing passes can be used to improve the contour and smoothness of DSAEK grafts without damaging donor endothelial cells. Clinical trials are needed to determine whether reshaping donors using excimer laser can deliver improved visual outcomes after DSAEK.

Purpose: The purpose of this study was to evaluate outcomes of Descemet's stripping automated endothelial keratoplasty (DSAEK) using anterior stromal flawed (ASF) donor corneas that were unsuitable for use in full-thickness penetrating keratoplasty as a result of stromal scars, pterygia, or previous corneal refractive surgery and to compare results with DSAEK using standard tissue., Methods: We conducted a review of our initial 42 (19 with 6-month follow up) consecutive DSAEK surgeries using ASF tissue compared with 357 (199 with 6-month follow up) time-matched controls using standard tissue. Intraoperative and perioperative complications, including dislocations and primary graft failures, were compared. Six-month best spectacle-corrected vision, incidence of rejection episodes, postoperative refractive astigmatism, keratometric values, pre- and postoperative topography-derived surface asymmetry index, and surface regularity index were compared., Results: One surgeon-cut ASF tissue was perforated before surgery and was discarded. No surgeon-cut standard tissue was perforated. No intraoperative complications and no episodes of primary graft failure or pupillary block glaucoma occurred in either group. One (2.4%) postoperative graft dislocation and one (5.2%) graft rejection episode occurred in the study group. There were 10 (2.8%) dislocations and 8 (2.2%) graft rejection in the controls. A statistically similar significant improvement in best spectacle-corrected vision occurred in both groups. Corneal topography, pachymetry, and manifest astigmatism were not significantly different between groups., Conclusion: Postoperative results of DSAEK using donor tissue excluded from use in penetrating keratoplasty as a result of stromal flaws are equivalent to results using standard donor tissue. Central corneal thickness measurements should be performed before cutting to avoid tissue perforation. The use of ASF tissue for DSAEK will expand the cornea donor pool.

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.

Purpose: We developed a simple, practical, and inexpensive technique to analyze areas of endothelial cell loss and/or damage over the entire corneal area after vital dye staining by using a readily available, off-the-shelf, consumer software program, Adobe Photoshop. The purpose of this article is to convey a method of quantifying areas of cell loss and/or damage., Methods: Descemet-stripping automated endothelial keratoplasty corneal transplant surgery was performed by using 5 precut corneas on a human cadaver eye. Corneas were removed and stained with trypan blue and alizarin red S and subsequently photographed. Quantitative assessment of endothelial damage was performed by using Adobe Photoshop 7.0 software., Results: The average difference for cell area damage for analyses performed by 1 observer twice was 1.41%. For analyses performed by 2 observers, the average difference was 1.71%. Three masked observers were 100% successful in matching the randomized stained corneas to their randomized processed Adobe images., Conclusions: Vital dye staining of corneal endothelial cells can be combined with Adobe Photoshop software to yield a quantitative assessment of areas of acute endothelial cell loss and/or damage. This described technique holds promise for a more consistent and accurate method to evaluate the surgical trauma to the endothelial cell layer in laboratory models. This method of quantitative analysis can probably be generalized to any area of research that involves areas that are differentiated by color or contrast.

Purpose: To evaluate the use of corneal donor tissue deemed unsuitable for full-thickness penetrating keratoplasty (PK) for use in deep lamellar endothelial keratoplasty (DLEK) and to compare postoperative results to those of DLEK surgery using donor tissue that is suitable for PK., Methods: Small-incision DLEK surgery was performed using 39 donor corneas unsuitable for PK. Thirty-five donors had anterior scars or opacities, 3 donors had pterygia within the 8-mm zone, and 1 had prior LASIK. All donor preparation was completed by manual stromal dissection. The DLEK surgical and postoperative courses were reviewed. Preoperative and 6-month postoperative results of this study group were compared with a control group consisting of the first 55 consecutive small-incision DLEK patients receiving donor corneas that had no criteria excluding them from use in PK. Four eyes in the study group and 1 eye in the control group had the confounding variables of the presence of an anterior-chamber lens or surgical vitrectomy with macular disease in the recipient eye., Results: There was no significant difference in preoperative measurements of best spectacle-corrected visual acuity (BSCVA; P = 0.372), donor endothelial cell density (ECD; P = 0.749), or corneal topography [surface regularity index (SRI), P = 0.485; or surface asymmetry index (SAI), P = 0.154] between the 2 groups. For the patients receiving corneas deemed unacceptable for PK, at 6 months after surgery, the vision (P = 0.002) and corneal topography measurements improved significantly from before surgery (SRI, P < 0.001; SAI, P < 0.001), and there was no significant change in refractive astigmatism (P = 0.240). There was a significant difference in the vision at 6 months postoperatively between the overall study group and the control group, with the mean vision of the study group at 20/56 and the control group at 20/43 (P = 0.015). If eyes with known cystoid macular edema (CME) and vitrectomy are removed from each group, there is no significant difference in vision at 6 months between the study group and the control group (P = 0.110), with the average BSCVA of those receiving donor corneas unsuitable for PK equal to 20/48 (range, 20/25-20/200) and the average vision for those receiving PK-acceptable donor tissue equal to 20/43 (range, 20/20-20/80). The 6-month average refractive astigmatism of the study group was 1.12 +/- 0.99 D (range, 0.00-4.00 D), and the average endothelial cell count was 2064 +/- 396 cells/mm(2) (range, 1208-2957 cells/mm(2)). There was no significant difference in 6-month postoperative endothelial cell count (P = 0.443), refractive astigmatism (P = 0.567), or corneal topography (SRI, P = 0.332; SAI, P = 0.110) in study patients who received corneas unsuitable for PK compared with control patients who received corneas suitable for PK., Conclusions: Endothelial keratoplasty such as DLEK surgery with manual donor preparation broadens the donor pool by enabling corneas that cannot be used for PK to be used for selective endothelial transplantation without deleterious postoperative results.