Correlation Between Optical Coherence Tomography and Photopic Negative Response of Flash Electroretinography in Ganglion Cell Complex Assessment in Glaucoma Patients.

Purpose: To investigate the correlation between the photopic negative response (PhNR) of the light-adapted flash electroretinography (ERG) and measurements of standard automated perimetry (SAP) and optical coherence tomography (OCT) in assessment of retinal ganglion cells' (RGCs) affection in glaucoma. Patients and Methods: A cross-sectional study included 40 eyes of glaucoma patients and 40 eyes of age- and gender-matched normal subjects. Participants underwent a complete ophthalmologic assessment, SAP, OCT, and light-adapted flash ERG using the extended PhNR protocol of the International Society for Clinical Electrophysiology of Vision (ISCEV). Glaucomatous eyes were divided into 3 subgroups: mild (n = 15), moderate (n = 11) and severe glaucoma (n = 14) according to the mean deviation (MD) of SAP. Measurements of SAP, OCT and ERG parameters were analyzed, and correlations between PhNR measurements and other study measurements were evaluated. Results: PhNR amplitudes and PhNR/b-wave ratios were significantly reduced in glaucoma cases compared to healthy controls, and they showed a significant and progressive decline across the three glaucoma subgroups (P < 0.05). An exception to this is PT (b-wave peak to PhNR trough) PhNR amplitude where its reduction was statistically non-significant when comparing between controls and mild glaucoma cases (P = 0.178), and between moderate and severe glaucoma cases (P = 0.714). PhNR amplitudes and PhNR/b-wave ratios correlated significantly with SAP and OCT parameters (P < 0.05). Conclusion: PhNR correlates well with SAP and OCT parameters in glaucoma assessment. PhNR could be a valuable supplementary tool for objective assessment of the RGCs' function in glaucoma.

Comparison of the pig and feline models for full thickness corneal transplantation.

PURPOSE: The goal of this study was to report on the advantages and limitations of the pig and feline models for experimental in vivo corneal transplantation. METHODS: Ten healthy domestic pigs and ten healthy cats were used. Full thickness penetrating keratoplasty was performed using autologous (eight cases), allogeneic (seven cases) or human xenogeneic (three cases) tissue. In two other cases, the inflammatory response to partial thickness trephination (without transplantation) was evaluated. Eyes were assessed daily before and after surgery by slit-lamp, pachymetry, and tonometry. A transparency score ranging from 0 (opaque graft) to 4 (clear graft) was used, based on the slit-lamp examination. Optical coherence tomography, histology, and electron microscopy were performed postmortem. RESULTS: In the pig, the mean (±SD) transparency score for the eight full thickness grafts was 0.88 ± 0.99, ranging from 0 to 3. In the feline model, the mean transparency score for the seven uncomplicated grafts was 3.93 ± 0.19, ranging from 3.5 to 4. Both negative controls without endothelium remained opaque at all time. Intraoperative tendency for iris incarceration into the wound, rapid corneal swelling, suture cheese wiring, and postoperative intraocular inflammation were the main factors jeopardizing the functional success of the corneal transplant in the pig model. CONCLUSION: Suboptimal functional results were obtained after full thickness corneal transplantation in the pig model, while in the feline model, the same protocol yielded uneventful surgeries and clear transplants, with functional results similar to those achieved in human subjects.

Effect of corneal hydration on the quality of the femtosecond laser anterior lamellar cut.

The goal of this study was to assess the effect of corneal hydration on the quality of the femtosecond laser (FSL) anterior lamellar cut. The Visumax FSL was used to dissect an 8-mm-diameter corneal flap in 22 eye bank corneas showing various levels of hydration. The intended ablation depth was 220 µm in all eyes, which corresponded to the maximal depth available with this laser. After the cut, the achieved ablation depth was measured using optical coherence tomography images, flap separability was assessed by measuring the mean force generated to detach the flap, and stromal bed roughness was assessed by measuring the Haralick contrast level on the 1000× scanning electron microscopy images of the ablated surfaces. The preoperative central corneal thickness ranged from 547 to 1104 µm (mean ± SEM: 833 ± 30 µm). A negative correlation was found between the level of corneal hydration and the ablation depth measured in the mid-peripheral cornea (r =  -0.626, p = 0.003), the ablation being more superficial in more edematous corneas. The Haralick contrast also tended to increase as a function of corneal hydration (r = 0.416, p = 0.061), suggesting that laser ablation in edematous corneas results in rougher stromal surfaces. These results support the hypothesis that the quality of the FSL lamellar cut decreases as the level of corneal hydration increases. Although FSL is still considered in the field as the tool of the future for corneal dissection, a better understanding of the limits of this tool will be needed before it can replace manual or automated stromal dissection techniques in hydrated corneas.

Smoothness assessment of corneal stromal surfaces.

PURPOSE: To assess the accuracy of the scanning electron microscopy (SEM) and present alternative approaches to quantify surface roughness based on numerical analysis. SETTING: Department of Ophthalmology, Maisonneuve-Rosemont Hospital, University of Montreal, Montreal, Quebec, Canada. DESIGN: Experimental study. METHODS: Lamellar stromal cuts were performed on human corneas using a femtosecond laser or a microkeratome. The photodisrupted stromal surfaces were processed for SEM, and images were acquired at ×1000 magnification. First, images were evaluated by independent observers. Second, images were analyzed based on first-order and second-order statistics of gray-level intensities. Third, 3-dimensional (3-D) surface reconstructions were generated from pairs of SEM images acquired at 2 angles. RESULTS: Results show that traditional assessment of roughness based on evaluating SEM images by independent observers can be replaced by computer-image texture analysis; an algorithm was developed to avoid subjective and time-consuming observations. The 3-D reconstructions allowed additional characterization of surface properties that was not possible with SEM images alone. Significant fluctuations in surface height were lost, although they could be retrieved using 3-D reconstructions. CONCLUSIONS: Image texture analysis allowed objective and repeatable assessment of stromal surface roughness; however, full assessments of surface-height fluctuations required 3-D reconstruction. These complementary methodologies offer a more comprehensive assessment of corneal surface roughness in clinical applications.

Corneal endothelial toxicity of air and SF6.

PURPOSE: The authors conducted in vivo assessment of corneal endothelial toxicity of air and SF6 in the feline model. This research was motivated by the increased use of air in anterior segment surgery in human subjects. METHODS: This was a prospective masked study. The eyes of 16 healthy adult cats were randomly assigned for the injection of 0.7 mL air into the anterior chamber of one eye and SF6 in the contralateral eye. Daily examination included slit lamp photographs, pachymetry, and tonometry. Specular microscopy was performed before, 7 days after, and 10 days after injection. The animals were euthanatized, and the corneas were processed for alizarin red-trypan blue staining and for light and electron microscopy. RESULTS: SF6 remained in the anterior chamber significantly longer than air. Both groups showed postinjection inflammation, which on average was maximal at day 2 and more severe with SF6. No difference in IOP was observed between the two groups. Specular microscopy showed significant endothelial cell loss in the SF6 group (mean postinjection cell loss, 132 ± 50 cells/mm(2)) but not in the group injected with air. Alizarin red staining revealed significant regional differences in cell density only in the SF6 group and more pronounced endothelial cell loss in the superior area. CONCLUSIONS: These results indicate that both air and SF6 injected into the anterior chamber of the eye can induce intraocular reaction in the feline model and that SF6 is more toxic than air in terms of endothelial cell loss and anterior chamber inflammation.

Optical coherence tomography anatomy of the corneal endothelial transplantation wound.

PURPOSE: The goal of this study was to prospectively assess the deep lamellar endothelial keratoplasty (DLEK) wound anatomy and its evolution during the 12 months after surgery, using optical coherence tomography (OCT). METHODS: The eyes of 8 patients (1 eye per patient) who consecutively underwent DLEK for Fuchs dystrophy or pseudophakic bullous keratopathy were prospectively studied before and 1, 3, 6, and 12 months after surgery. The Stratus OCT apparatus (Carl Zeiss Meditec, Dublin, CA) was used to acquire central and radial scans perpendicular to the wound at 3-, 6-, 9-, and 12-o'clock positions. The following parameters were analyzed: central total thickness, posterior donor-recipient edges gap, donor-recipient height mismatch, tissue compression, and graft detachment. RESULTS: A posterior gap was observed in 4 of the 8 DLEK eyes. At 12 months, the mean gap contour, depth, and width were 242 +/- 67, 101 +/- 45, and 87 +/- 29 microm, respectively. A step was documented in all DLEK eyes (average step height 108 +/- 24 microm). A micrograft detachment was observed in one case and tissue compression in another. In all corneas, the mean central corneal thickness returned to normal range and almost normal anatomy with time after surgery. CONCLUSIONS: OCT was found to be a very useful tool for DLEK corneal wound architecture analysis. It revealed microscopic wound irregularities and allowed their quantitative follow-up with time.

Transplantation of a tissue-engineered corneal endothelium reconstructed on a devitalized carrier in the feline model.

PURPOSE: To evaluate the functional outcome of tissue-engineered corneal endothelium reconstructed on a devitalized carrier and transplanted in the living feline model. METHODS: Eighteen healthy adult cats underwent full-thickness corneal transplantation. In 11 animals, the donor cornea was reconstructed from cultured allogeneic feline corneal endothelial cells seeded on the denuded Descemet's membrane of a devitalized human cornea. The reconstructed corneal endothelium was cultured for 2 weeks before transplantation. Five control animals received autologous (n = 1), allogeneic (n = 3), or human xenogeneic (n = 1) native cornea. Two other control animals were grafted with the devitalized carrier only (no cells). Animals were observed daily by slit lamp until euthanatization on day 7. Postmortem analysis included optical coherence tomography (OCT), alizarin red staining, histology, fluorescence microscopy, scanning electron microscopy (SEM), and transmission electron microscopy (TEM). RESULTS: Nine of the 11 reconstructed corneal endothelial grafts and all five native (autologous, allogeneic, xenogeneic) control grafts were clear and thin 7 days after grafting. In contrast, the two control grafts consisting of the carrier only (without endothelium) remained thick and opaque. Alizarin red staining, histology, SEM, and TEM showed that the transplanted reconstructed endothelium maintained a normal morphology and ultrastructure and expressed the function-related proteins Na(+)/K(+)-ATPase alpha1, Na(+)/HCO(3), and ZO-1. CONCLUSIONS: This study provides evidence for the short-term (7-day) anatomic and functional success of corneal transplantation with a tissue-engineered corneal endothelium reconstructed on a devitalized carrier.

Laser ablation threshold dependence on pulse duration for fused silica and corneal tissues: experiments and modeling.

The surface ablation threshold fluence of fused silica and two porcine cornea layers, the epithelium and the stroma, is characterized as a function of the laser pulse duration in the range of 100 fs-5 ps for a wavelength of 800 nm (Ti:sapphire laser system). The plateaulike region observed between 100 fs and 1 ps for the corneal layers indicates that for use in laser surgery, laser pulse durations chosen within this range should be practically equivalent. Our model predicts that the ablation threshold will decrease rapidly for pulse durations in the low end of the femtosecond regime.