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Retinal Ganglion Cell and Inner Plexiform Layer Thickness Measurements AL De a Moura Et Al 1189 Eye (2012) 26, 1188–1193 & 2012 Macmillan Publishers Limited All rights reserved 0950-222X/12 www.nature.com/eye 1;2;3 1 1 CLINICAL STUDY Retinal ganglion cell AL de A Moura , AS Raza , MA Lazow , CG De Moraes4;5 and DC Hood1;6 and inner plexiform layer thickness measurements in regions of severe visual field sensitivity loss in 1Department of Psychology, patients with Columbia University, New York, NY, USA glaucoma 2Department of Experimental Psychology, University of Sa˜ o Paulo, Sa˜ o Paulo, Brazil Abstract Purpose To better understand the effects of RGC layer thickness exceeds the IP layer 3Department of Ophthalmology, ABC School severe glaucoma on the thickness of the retinal thickness in controls. of Medicine, Sa˜ o Paulo, ganglion cell (RGC) and inner plexiform (IP) Eye (2012) 26, 1188–1193; doi:10.1038/eye.2012.110; Brazil layers measured with frequency-domain optical published online 15 June 2012 coherence tomography. 4Einhorn Clinical Research Methods In experiment 1, macular cube Keywords: glaucoma; OCT; ganglion cell layer; Center, New York Eye and scans were obtained in 11 patients with inner plexiform layer Ear Infirmary, New York, NY, USA glaucoma and the thickness of both the RGC and IP layers were measured at locations Introduction 5Department of corresponding to 3, 5, and 71 eccentricity. For Ophthalmology, New York patients, only locations with total deviation Glaucomatous optic neuropathy has been University School of losses of À 15 dB or worse on perimetry were traditionally diagnosed based upon structural Medicine, New York, included. In experiment 2, higher resolution, (optic disc appearance) and functional (visual NY, USA horizontal midline scans were obtained from field (VF)) abnormalities. In the last 10 years, 6Department of 30 controls in order to obtain a precise imaging techniques have allowed for structural Ophthalmology, Columbia measure of the thickness of the RGC and measurements of the nerve fiber rim and retinal University, New York, NY, IP layers of the healthy retina. nerve fiber layer (RNFL) thicknesses and for USA Results In regions of severe field loss quantitative comparisons of structural thickness (experiment 1), glaucoma decreased the to VF loss.1–8 However, because these Correspondence: thickness of both layers, leaving a residual techniques measure RNFL thickness, they AL de A Moura, Departamento de layer. The residual thickness of the IP layer provide an indirect measure of local retinal Psicologia, Universidade de was larger than the residual thickness of ganglion cell (RGC) damage. Sa˜ o Paulo, Av Prof. Mello the RGC layer. In healthy controls With the advent of the higher resolution Moraes, 1721, Bloco B–D9, (experiment 2), the RGC layer was about frequency-domain optical coherence Sa˜ o Paulo 05508-030, 57% of the RGC þ IP layer thickness at 31 as tomography (fdOCT), it became possible to Brazil 1 Tel: +55 11 30914263; compared with only 36% at 10 ,in measure local RGC thickness in all the 9,10 Fax: +55 11 30914357; agreement with a recent histological study. important macular region. In a recent study, E-mail: almoura_oft@ Conclusion Glaucomatous optic neuropathy, Raza et al11 found a good correlation between yahoo.com.br with severe losses in visual field sensitivity, local RGC plus inner plexiform (IP) layer decreases the thickness of both the RGC and thickness and local loss of VF sensitivity in Received: 8 November 2011 IP layers, but leaves a residual thickness of patients with glaucoma, within about central 81 Accepted in revised form: 30 April 2012 both. The IP layer contributes slightly more of fixation. Interestingly, the RGC þ IP layer Published online: 15 June than the RGC to this residual, even just thickness reached an asymptotic thickness of 2012 outside the center of the fovea where the about 45 mm, no matter how extreme the field Retinal ganglion cell and inner plexiform layer thickness measurements AL de A Moura et al 1189 loss was. The authors suggested that this relatively large ophthalmological examination and VF. Each patient was residual, which was about one-half of the RGC þ IP layer invited to take part in the study only if the inclusion at its thickest point, might be due to a relative criteria was satisfied, which included the presence of preservation of the IP layer. severe VF loss ( À 15 dB) inside the central 101, reliable Raza et al,11 however, could not test this hypothesis, as VFs and absence of systemic diseases or any other it was not possible to reliably discern separate RGC and ophthalmological pathology besides glaucoma. Results IP layers in their images. In the present study, we will were compared with 20 age-matched, healthy controls make use of higher resolution scans. In experiment 1, with no ophthalmological or systemic diseases. we analyzed local changes in both RGC and IP layer thicknesses in patients with severe VF losses secondary to glaucoma, scanned under conditions allowing for Equipment and procedures better visualization of both the RGC and IP layers. Standard automated perimetry. VFs were performed To better understand how the relative thickness of the using the Humphrey Field Analyzer program 24-2 SITA RGC and IP layers varies with eccentricity in healthy Standard (Carl Zeiss Meditec, Dublin, CA, USA). The eyes, we also examined the thickness of these layers in total deviation (TD) score, which is the difference in dB in higher resolution horizontal OCT line scans. The the patient’s sensitivity when compared with an age- results of experiment 2 will be compared with a recent matched control group, was used for the analysis. The histological study of the postmortem human retina.12 16 field points of the 24-2 pattern within the central 101 were examined to identify the regions with a TD value Materials and methods equal to À 15 dB or worse. Experiment 1 OCT. All the individuals were scanned with an fdOCT Retinal layers from patients with glaucoma were studied (Spectralis HRA-OCT; Heidelberg Engineering, Vista, with cube scans and an eye-tracking system, which CA, USA) using the eye-tracking feature and a three- improved the definition between RGC and IP layers. To dimensional/volume macular scan protocol, which directly measure the relative loss in RGC and IP layers consisted of 128 horizontal lines (B-scans) across a due to glaucoma, regions with severe losses in field 6 Â 6mm2 area, a protocol routinely employed in our sensitivity and clearly discernable separate RGC and IP clinic. These scans, centered on the fovea, were about 201 layers were examined. in height and width and had images of the necessary quality, that is, with no blinking or movement artifacts, for the segmentation procedure described below. Participants The OCT images were exported to a program written We tested 11 patients with glaucoma (mean age in Matlab 711 (MathWorks, Natick, MA, USA). The retinal 65±8.21 years) (Table 1) who were diagnosed based on layers were segmented with a computer-aided, manual Table 1 Demographic data of the patients with glaucoma Patient no Code Age Gender Eye VA IOP TD (101) MD PSD TG 1 7343 62 M OS 20/30 8 À 13.18 À 16.90 13.62 POAG (NTG) 2 7467 72 M OD 20/25 13 À 3.87 À 2.95 7.35 POAG 3 7481 59 M OD 20/15 21 À 13.18 À 12.07 14.16 POAG 4 7494 64 F OD 20/20 14 À 7.62 À 7.68 8.37 POAG 5 7673 61 F OS 20/20 12 À 15.81 À 18.39 13.89 POAG 6 7755 61 F OD 20/15 13 À 17.06 À 13.03 16.68 POAG (NTG) 7 7811 81 F OS 20/25 10 À 9.25 À 6.51 8.99 POAG 8 7831 76 F OS 20/30 15 À 28.00 À 26.68 10.00 XFG 9 7269 52 M OD 20/30 14 À 19.31 À 14.71 13.79 POAG 10 7245 64 F OS 20/20 12 À 29.68 À 29.10 6.46 POAG 11 7322 66 M OD 20/25 10 À 10.93 À 10.68 10.45 POAG Mean 65.27 12.91 À 15.26 À 14.43 11.25 SD 8.21 3.39 À 8.01 À 8.05 3.33 Abbreviations: F, female; IOP, intraocular pressure (at the time of the visual field evaluation and OCT measurements); M, male; MD, mean deviation; NTG, normal tension glaucoma; POAG, primary open angle glaucoma; PSD, pattern standard deviation; TG, type of glaucoma; TD, total deviation (inside 10 degrees); VA, visual acuity; XFG, exfoliative glaucoma. Eye Retinal ganglion cell and inner plexiform layer thickness measurements AL de A Moura et al 1190 high-resolution line scans averaged across the horizontal meridian. Participants For this experiment, 30 eyes from 30 healthy controls were studied (35.7±14.0 years); 20 of these eyes were part of a control group previously published.13 All the participants satisfied the inclusion criteria for controls, which were absence of ophthalmological or systemic diseases. Participants included in experiment 2 were not the same as those included in experiment 1. Equipment and procedures OCT. All the individuals were scanned with a fdOCT Figure 1 Macular locations of the scans at 0, 3, 5, and 71. (Spectralis HRA-OCT; Heidelberg Engineering) using the eye-tracking feature. To obtain high-resolution scans, segmentation procedure,9,13 previously shown to 100 horizontal line scans taken through the fovea were produce reliable and reproducible results.14 The images averaged. The exported scans were segmented as corresponding to the seven scan lines were analyzed described in experiment 1, and measurements of the (Figure 1).
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