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In Vitro Corneal Tomography of Donor Cornea Using Anterior Segment OCT

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In Vitro Corneal Tomography of Donor Cornea Using Anterior Segment OCT BASIC INVESTIGATION In Vitro Corneal Tomography of Donor Cornea Using Anterior Segment OCT Edgar Janunts, PhD,* Achim Langenbucher, PhD,* and Berthold Seitz, ML, MD† n cases of corneal donation, either the entire globe or just Purpose: The aim of this study was to establish a tomographic Ia 15-mm corneoscleral piece of tissue is taken. Having screening method for revealing potential pathologies in corneal confirmed the absence of obvious scars on the cornea and donors before keratoplasty so they may be excluded as candidates for a medical history without any corneal surgeries or infectious corneal transplantation. diseases, the corneoscleral button can be excised and stored in fi “ ” Methods: Donor corneal tomographies were measured in a viewing a special chamber lled with a transport medium before chamber filled with preservation medium and with the use of transplantation. At present, only a visual examination using a clinical optical coherence tomography (OCT) device. Custom- a handheld slit lamp is performed before the corneoscleral written software was developed to extract corneal surfaces from the excision, in addition to blood and smear tests. Further slit- raw data, which were analyzed in the central and peripheral regions. lamp examinations and microbiological tests are usually An adaptive nonlinear edge-enhancement algorithm was used to performed under laboratory conditions in eye banks. There observe scars within the corneal volume. The thickness distribution is no tomographic screening routine presently available that map was analyzed to detect keratoconus and corneas with extreme discounts corneas that might be unsuitable for future trans- topographic irregularities. Measurements were repeated 5 times to plant, such as those which have suffered previous surgeries assess reproducibility. (eg, LASIK) or diseases (eg, keratoconus) that might affect the biomechanical properties of the cornea. Results: Eight corneas were investigated: 6 randomly selected There are a few case reports in the literature in which, intact donors, unsuitable for implantation because of low endothelial despite the routine examinations mentioned above, donated cell densities, and 2 keratoconus corneas, excised from patients corneas, which had previously undergone an operation, were during corneal transplantation. A major thickness abnormality was accepted for transplantation. Two clinical cases were reported1 detected in one of the intact donor corneas, so it was excluded from where corneas were only diagnosed as having undergone further analysis. The keratoconus corneas were clearly evident in a LASIK procedure after they had been transplanted. Ehren- optical coherence tomography cross-sectional images, and similarly, haus et al2 reported a case of a donor cornea with a soft contact fi they could easily be identi ed by analyzing the thickness map. lens still in place. A recent study by Kang et al3 estimated that ’ Overall, the measurements were reliable and had a Cronbach s alpha approximately 10% of corneas in eye banks have undergone an fi coef cient greater than 0.8. as yet unidentified LASIK procedure. – Conclusions: Donor corneal examination using sterile viewing In the past, Placido ring based video keratographs were 4,5 chambers was found to be suitable as a pre-keratoplasty advanced adapted for use under eye bank conditions, whereas more screening routine. A proof of concept was demonstrated, which recent studies favor the screening of donor corneal tissue with 6,7 could identify both irregular corneas and those affected the use of optical coherence tomography (OCT). Ousley 4 by keratoconus. and Terry introduced a portable, Placido-based topography instrument, known as a Keratron Scout, for donor screening. Key Words: eye bank, donor corneal examination, keratoconus, The authors also proposed a method for the identification of keratoplasty, tomographic screening tissues that had undergone refractive surgery by observing the (Cornea 2016;35:647–653) difference in corneal curvature at the 7- and 3-mm zones. They showed that corneal curvature does not change much before and after excision from the donor; however, moderate changes in astigmatism could be observed.4 Lin8 developed a laboratory OCT system to perform Received for publication August 4, 2015; revision received November 23, corneal screening in a viewing chamber; the authors used 2015; accepted December 13, 2015. Published online ahead of print a custom-made high-resolution laser source and monitored March 1, 2016. From the *Institute of Experimental Ophthalmology, University of Saarland, refractive index distributions to differentiate between normal Homburg, Saar, Germany; and †Department of Ophthalmology, Saarland corneas and LASIK-operated corneas. In a different work, Lin University Medical Center, Homburg, Saar, Germany. et al9 proposed a method to detect LASIK-operated corneas, The authors have no funding or conflicts of interest to disclose. which involved measuring the anterior curvature and the Reprints: Edgar Janunts, PhD, Institute of Experimental Ophthalmology, fl Saarland University, Kirrberger St 100, Bldg 22, 66421 Homburg/Saar, anterior/posterior stromal re ectivity ratio with OCT scans. Germany (e-mail: [email protected]). The present study aims to propose a tomographic screening Copyright © 2016 Wolters Kluwer Health, Inc. All rights reserved. method to detect possible pathologies in donor material and Cornea Volume 35, Number 5, May 2016 www.corneajrnl.com | 647 Copyright Ó 2016 Wolters Kluwer Health, Inc. Unauthorized reproduction of this article is prohibited. Janunts et al Cornea Volume 35, Number 5, May 2016 exclude these potential donors from future transplantations data processing. The image processing software automatically (among others, corneas that have undergone refractive detected the cornea and corrected the images by compensat- surgery and those with geometrical abnormalities, ing the refraction error of the scanning beam passing through eg, keratoconus). layers with different refractive indices [air, cuvette (glass), liquid, cornea, and liquid]. MATERIALS AND METHODS Image Processing Algorithm The donor corneal examination was performed using The contrast of the OCT images was enhanced at first, a viewing chamber from Bausch and Lomb by an AS-OCT then the images were smoothed using a standard Gaussian SS-1000 CASIA clinical tomographer (Tomey) in a laboratory filter, and the noise was removed using LabVIEW’s particle environment without removing the corneas from the sterile analysis algorithm. The corneal surfaces were then extracted preservation medium (Fig. 1). with Canny edge filtering and presented as separate surfaces. The viewing chamber was filled with transport medium, Finally, the surfaces were corrected according to Snell’s law a commercial product from Biochrom that consists of the (Fig. 2), and the thickness map was computed. following substances: MEM Earle’s salt (·10), penicillin/ streptomycin, L-glutamine (200 mM), amphotericin B (250 mg/mL), HEPES buffer (1 M), and NaHCO3. Methods of Analysis After measuring the cornea, a custom-written software Based on the anterior surface and the pachymetry maps using LabVIEW 2013 (National Instruments) was applied for (Figs. 3, 4), various automatic analyses were programmed to FIGURE 1. A 3-dimensional cross-sectional examination of donor corneoscleral tissue using an SS-1000 CASIA clinical OCT to- mographer (Tomey). 648 | www.corneajrnl.com Copyright © 2016 Wolters Kluwer Health, Inc. All rights reserved. Copyright Ó 2016 Wolters Kluwer Health, Inc. Unauthorized reproduction of this article is prohibited. Cornea Volume 35, Number 5, May 2016 Donor Corneal Tomography Using Anterior Segment OCT FIGURE 2. Image processing algorithm. The contrast of the raw image [OCT cross-sectional scan (A)] was enhanced and a standard Gaussian filter applied (B). Particle analysis was then carried out to reduce noise, thus removing all particles unrelated to the cornea (C). In subsequent steps, the corneal volume was separated by applying Canny edge filters (D). Finally, both corneal surfaces were recognized (E) and extracted as separate curves (F). identify donor corneas that may potentially be unacceptable Furthermore, in addition to the adaptive edge-enhanced for keratoplasty. approach, a mathematical evaluation model was used to In particular, a mathematical model was used to diagnose the presence of postrefractive surgery corneas by diagnose scars in the corneal tissue, such as flaps resulting comparing the difference in anterior corneal curvature of the from LASIK surgery, intracorneal rings, or femtosecond central (3 mm) and peripheral (7 mm) regions, as proposed by laser–assisted keratoplasties, by using an adaptive edge- Ousley and Terry.4 Both spherical and aspherical parameters enhancement algorithm. Here a color-coded cross-sectional were analyzed. image was computed (Fig. 3), where brighter coloring To detect corneas with keratoconus disease or thickness indicated a higher contrast. The algorithm comprises a local abnormalities, analysis of entire thickness distribution of the filtering method based on a Taylor expansion.10 The filtered cornea was performed by superimposing all cross-sectional images were smoothed using Gaussian smoothing followed pachymetries (B-scans) onto a 2-dimensional graph, as shown by a nonlinear filter for detecting local discontinuities. in Figs. 4–7. This new approach defines the thickness of the cornea in the center and then the maximum difference in thickness with respect
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