Intraoperative use of spectral-domain optical coherence tomography during Descemet's stripping automated endothelial keratoplasty.

PURPOSE: To evaluate the intraoperative changes in the donor lenticule, recipient cornea, and the reduction of interface fluid thickness during Descemet's stripping and automated endothelial keratoplasty with EndoGlide™ (Angiotech Pharmaceuticals Inc, Vancouver, Canada) donor insertion, using intraoperative spectral-domain optical coherence tomography. METHODS: Prospective observational case series of patients underwent Descemet's stripping and automated endothelial keratoplasty using the EndoGlide inserter. Spectral-domain optical coherence tomography (iVue; Optovue Inc, Fremont, CA) with a handheld probe was used to image the cornea and anterior chamber. Standardized software was used to measure interface fluid gap, host cornea, and donor lenticule thicknesses during the following surgical stages of Descemet's stripping and automated endothelial keratoplasty: (1) after donor insertion and immediately before full air tamponade; (2) after air tamponade and expression of fluid from venting incisions; (3) at 6 minutes of air tamponade; and (4) at 10 minutes of air tamponade. RESULTS: Ten patients with a mean age of 74.9 ± 11.8 years were recruited. Spectral-domain optical coherence tomography measurements of the interface fluid gap after fluid was expressed through the venting incisions (P < 0.001), at 6 minutes of air tamponade (P < 0.001) and at 10 minutes of air tamponade (P < 0.001 and P = 0.001, respectively), were significantly decreased compared to the measurements immediately before air tamponade. Donor thickness increased significantly at 6 minutes of air tamponade (P = 0.004) but reduced by 10 minutes compared to immediately before air tamponade. CONCLUSION: Significant intraoperative changes in the donor, recipient cornea, and interface fluid thickness occurred following endothelial keratoplasty donor insertion.

Evolution of deep anterior lamellar keratoplasty (DALK).

The concept of selective tissue transplantation was theorized over two centuries ago. However, deep anterior lamellar keratoplasty (DALK) was difficult to perform with the instruments available at that time, and visual acuity results were poor compared to those achieved by full-thickness transplantation. Recently, many technical improvements in surgical instrumentation and advances in eye bank technology have led to the re-emergence of DALK. Now, patients with stromal pathologies not affecting the corneal endothelium can benefit from this surgery, which allows total stromal replacement of the recipient's cornea, leaving an intact recipient Descemet membrane and endothelium. Along with the advances in surgical instrumentation, improvements in surgical techniques and imaging technology have contributed to improved visual outcomes obtained with DALK. DALK is now seen as a viable alternative to penetrating keratoplasty with equivalent visual results and better long-term graft survival.

Comparison of eye-tracking success in laser in situ keratomileusis after flap creation with 2 femtosecond laser models.

PURPOSE: To determine the efficiency of an eye tracker after laser in situ keratomileusis (LASIK) flap creation with 1 of 2 femtosecond laser models. SETTING: Tertiary referral center, Singapore National Eye Center, Singapore. DESIGN: Randomized clinical trial. METHODS: The LASIK flap was created with an IntraLase (Group A) or a VisuMax (Group B) femtosecond laser. An Advanced Control Eye Tracker was initiated 3 times to obtain iris recognition. Eye tracking was considered successful if the eye movements could be followed despite the presence or absence of an opaque bubble layer (OBL). Univariate-multivariate logistic regression analysis was performed. RESULTS: Preoperatively, the mean values (ranges) of the 87 eyes were sphere, -5.64 diopters (D) ± 2.17 (SD) (-9.75 to -0.25 D); cylinder 1.65 ± 1.63 D (-3.75 to 0.00 D); optical zone, 6.34 ± 0.20 mm (5.6 to 7.0 mm); keratometry, 43.48 ± 1.32 D (40.1 to 42.8 D); flap thickness, 109.30 + 5.21 µm (90 to 115 µm). Eye tracking was successful in 38 (90.5%) of 42 eyes in Group A and 43 (95.6%) of 45 eyes in Group B. No specific type of OBL was seen in either group. The LASIK was completed in all eyes. There was a statistically significant association between positive tracking and a smaller optical zone (P=.03). There were no statistically significant differences in eye tracking between the 2 femtosecond lasers. CONCLUSIONS: Eye tracking was achieved in more than 90% of cases after LASIK flap creation with 1 of 2 femtosecond laser models, even in the presence of an OBL.

In vivo real-time intraocular pressure variations during LASIK flap creation.

PURPOSE: To monitor and compare in vivo real-time intraocular pressure (IOP) in rabbit eyes undergoing LASIK flap creation using microkeratome and femtosecond laser. METHODS: Thirteen rabbit eyes in each group underwent LASIK flap creation using a microkeratome and a femtosecond laser. In vivo real-time IOP profile was measured using a 30-gauge needle with an IOP catheter sensor inserted into the anterior chamber from the limbus during surgery. RESULTS: In vivo real-time IOP monitoring was achieved in all cases, showing IOP variations during different phases of LASIK flap creation from docking of the instrument, start of surgery to the end of procedure, and monitoring the post-LASIK stabilization. IOP fluctuations were significantly lower in corneal flaps made with the femtosecond laser than with the microkeratome during globe suction (81.78+/-10.55 vs. 122.51+/-16.95 mm Hg), cutting (62.25+/-3.28 vs. 141.02+/-20.46 mm Hg), and suction (41.40+/-2.99 vs. 89.30+/-12.15). In contrast, femtosecond laser requires double the time (19+/-2 vs. 10+/-2 seconds for globe suction and 19+/-2 vs. 9+/-2 seconds for cutting) for completion of the procedure. CONCLUSIONS: The authors describe an accurate and reliable setup to measure and record in vivo real-time changes in IOP measurement from the anterior chamber during laser surgery. Femtosecond laser flap creation exerts less extreme IOP fluctuations with improved chamber stability but requires more procedure time than does microkeratome.

One-year follow-up of epithelial corneal cell sheet allografts mounted on platelet poor plasma in rabbits.

PURPOSE: To evaluate the usefulness of epithelial corneal sheets mounted on platelet poor plasma (PPP) for allograft transplantation of rabbits with total limbal stem cell deficiency (LSCD) and to prove its efficacy at 1 year after surgery. METHODS: LSCD was induced in 21 female rabbits by mechanical keratectomy. To configure the grafts, limbal biopsies were taken from male rabbits and cells were cultured on a fibroblast feeder layer grown on clotted autologous PPP. After keratectomy, grafts were sutured over the stroma. Control groups consisted of no implant or an implant of clotted PPP. Rabbits were euthanized at 3 and 12 months. Corneas and cultured sheets were processed for histopathology and immunohistochemistry (K3/12 and K19). Gender analysis was performed at 4 and 7 months. RESULTS: One rabbit had endophthalmitis, and another died of no apparent cause. The rest of the animals treated had no inflammation, showed a stratified epithelium, keratin 3/12 expression, and no expression of keratin 19. At 1 year, seven of eight rabbits showed no LSCD or corneal rejection signs. Y chromosomes were detected at 4 and 7 months postoperatively. All controls showed LSCD signs, erratic epithelium, and minimal cell differentiation; they revealed a slight expression of K3/12 and an expression of K19 in patchy patterns. CONCLUSIONS: Allografts contributed to restoring a healthy eye surface without signs of graft rejection. This technique seems to be a promising procedure for bilateral ocular surface diseases and may be useful for new therapeutic strategies.

Advances in corneal stem-cell transplantation in rabbits with severe ocular alkali burns.

PURPOSE: To evaluate the efficacy of autologous corneal epithelial sheet implantation in restoring transparency of rabbit corneas severely injured by alkaline and the effect of photocoagulation in arresting corneal neovessel ingrowth. SETTING: Ophthalmology Department, School of Biomedical Sciences, Universidad Austral, Buenos Aires, Argentina. METHODS: Limbal stem-cell deficiency (LSCD) was induced in 14 rabbits by alkali burns. A limbal cell biopsy was done in the contralateral eye, and the cells were cultured on a fibroblast feeder layer grown on autologous clotted platelet-poor plasma or commercial fibrin for 21 days. Anterior keratectomy was followed by suturing corneal cell sheets over the stroma. If regrowth of vessels occurred, argon laser photocoagulation was applied to them. Rabbits were killed at 30, 60, 90, 180, and 360 days and the corneas processed for histopathology and inmunohistochemistry. RESULTS: A small (2.5 mm(2)) limbal biopsy achieved stem-cell replication in vitro. Corneal clarity and epithelial defects evolved with a trend toward improvement. There was a significant reduction in corneal neovascularization. Histology showed a multilayered stratified epithelium including several epithelial-like cells with clear cytoplasm in the deepest part. There were no signs of intraepithelial mucin cells on the implanted corneas. Immunohistochemical results showed expression of cytokeratins 3 and 12 in the central corneal epithelium and an absence of cytokeratin 19. CONCLUSIONS: Autologous limbal epithelial cell transplantation improved the corneal surface in eyes with LSCD. Photocoagulation of neovessel ingrowth was effective over the 1-year follow-up. Results may facilitate the application of this technique in patients.

Preparation of platelet-rich plasma as a tissue adhesive for experimental transplantation in rabbits.

PURPOSE: Platelet-rich plasma (PRP) is an autologous substance with adhesive properties. We aimed at developing and testing the efficacy of a method for PRP preparation in rabbits. MATERIALS AND METHODS: An in vitro study was carried out to obtain PRP from forty rabbits and to analyze the number of platelets and type of substance needed to trigger platelet activation. To induce platelet activation, 5%, 10%, 25% and 50% CaCl solutions were used. Then, an in vivo study was performed in twelve rabbits to test PRP adhesiveness in lamellar corneal graft. A control group made up of six rabbits underwent corneal transplantation without using PRP. RESULTS: 5% CaCl was the most effective concentration in activating PRP, with a mean time of 19 minutes. An attached corneal flap was seen 3 months after surgery. A detached corneal button was seen in all controls. CONCLUSION: Our method was able to produce rabbit-derived PRP with suitable properties for soft tissue adhesion. These results could be useful for researchers of the growing fields of tissue repair and experimental transplantation.