Evaluation of inner retinal layers in patients with multiple sclerosis or neuromyelitis optica using optical coherence tomography.

PURPOSE: To evaluate the thickness of the inner retinal layers in the macula using frequency-domain optical coherence tomography (fd-OCT) in patients with demyelinating diseases. DESIGN: Cross-sectional study. PARTICIPANTS: A total of 301 eyes of 176 subjects were evaluated. Subjects were divided in 5 different groups: controls, neuromyelitis optica (NMO), longitudinally extensive transverse myelitis (LETM), multiple sclerosis with a history of optic neuritis (MS-ON), and multiple sclerosis without a history of optic neuritis (MS non-ON). METHODS: The individual layers from macular fd-OCT cube scans were segmented with an automated algorithm and then manually hand-corrected. For each scan, we determined the thickness of the retinal nerve fiber layer (RNFL), the combined retinal ganglion cell and inner plexiform layers (RGCL+), and the inner nuclear layer (INL). MAIN OUTCOME MEASURES: Macular RNFL, RGCL+, and INL thickness. RESULTS: The RNFL was significantly thinner than in controls for all patient groups (P ≤ 0.01). Macular RGCL+ thickness was significantly thinner than in controls for the NMO, MS-ON, and MS non-ON groups (P<0.001 for the 3 groups). The INL thickness was significantly thicker than in controls for the patients with NMO (P = 0.003) and LETM (P = 0.006) but not for those with MS-ON or MS non-ON. Although the RNFL and RGCL+ were not significantly different between the NMO and MS-ON groups, the patients with NMO had a significantly thicker INL than the patients with MS-ON (P = 0.02). CONCLUSIONS: Macular RNFL and RGCL+ demonstrate axonal and neural loss in patients with MS, either with or without ON, and in patients with NMO. In addition, the INL thickening occurs in patients with NMO and patients with LETM, and study of this layer may hold promise for differentiating between NMO and MS.

Pattern electroretinogram in neuromyelitis optica and multiple sclerosis with or without optic neuritis and its correlation with FD-OCT and perimetry.

PURPOSE: To evaluate the ability of transient pattern electroretinogram (PERG) parameters to differentiate between eyes of patients with neuromyelitis optica (NMO), longitudinally extensive transverse myelitis (LETM), multiple sclerosis with optic neuritis (MS + ON), multiple sclerosis without optic neuritis (MS - ON), and controls, to compare PERG and OCT with regard to discrimination ability, and to assess the correlation between PERG, FD-OCT, and visual field measurements (VFs). METHODS: Visual field measurements and full-field stimulation PERGs based on both 48- and 14-min checks were obtained from patients with MS (n = 28), NMO (n = 20), LETM (n = 18), and controls (n = 26). In addition, FD-OCT peripapillary retinal nerve fiber layer (RNFL) and segmented macular layer measurements were obtained and their correlation coefficients were determined. RESULTS: Compared to controls, PERG amplitude measurements were significantly reduced in eyes with NMO and MS + ON, but not in eyes with LETM and MS - ON. PERG amplitudes were significantly smaller in NMO and MS + ON eyes than in MS - ON eyes. PERG and OCT performance was similar except in NMO eyes where macular thickness parameters were more efficient at detecting abnormalities. A significant correlation was found between N95 amplitude values and OCT-measured macular ganglion cell layer thickness, total retinal thickness, and temporal peripapillary RNFL thickness. PERG amplitude was also significantly associated with VF sensitivity loss. No statistically significant difference was observed with regard to the best-performing parameters of the two methods. CONCLUSIONS: Pattern electroretinogram measurements were able to detect RNFL loss in MS + ON and NMO eyes, with a performance comparable to OCT. PERG amplitude measurements were reasonably well correlated with OCT-measured parameters.

Abnormal multifocal ERG findings in patients with normal-appearing retinal anatomy.

To evaluate eyes with abnormal visual fields and multifocal electroretinograms (mfERGs) but normal-appearing frequency-domain optical coherence tomography (fdOCT) scans, the thicknesses of the outer retinal layers were measured. A total of 25 eyes from 17 patients, including 15 eyes previously tested (Dale et al. in Doc Ophthalmol 120(2):175-186, 2009) were examined. All patients were evaluated with standard automated perimetry (SAP) using the 24-2 and/or 10-2 program (Zeiss Meditec), mfERG with 103 hexagons (Veris, EDI), and fdOCT imaging (3DOCT-2000, Topcon) with scans of the macula. All patients had reliable visual fields showing macular defects and good quality mfERG and fdOCT results. The mfERG results were classified as abnormal based on decreased amplitudes and/or increased latencies corresponding to the abnormal visual field. Based on visual inspection, three experienced observers classified the fdOCT scans as normal or inconclusive, as opposed to clearly abnormal. Retinal layers of the fdOCT scans were manually segmented with the aid of a computer program and compared to mean thicknesses from 20 controls. The thicknesses of the outer segment plus retinal pigment epithelium, total receptor, and inner nuclear layers were measured. Quantitative analysis of fdOCT scans demonstrated thinning of the outer retina in some scans that was not readily apparent on visual inspection. One or more of the outer retinal layers was significantly thinner in 15 of the 25 eyes. The absence of significant thinning in the other 10 eyes represents instances in which functional loss measured by visual fields and mfERGs can precede clear structural changes on fdOCT.

Beta-zone parapapillary atrophy and multifocal visual evoked potentials in eyes with glaucomatous optic neuropathy.

We investigated changes in multifocal visual evoked potential (mfVEP) responses due to beta-zone parapapillary atrophy (ßPPA). Patients with glaucomatous optic neuropathy (GON) with or without standard achromatic perimetry (SAP) abnormalities were referred for mfVEP testing during a 2-year period. Eyes with good quality optic disc stereophotographs and reliable SAP results were included. The mfVEP monocular mean latency delays (ms) and amplitudes (SNR) were analyzed. Age, SAP mean deviation (MD), pattern standard deviation (PSD), and spherical equivalent (SE) were analyzed in the multivariate model. Generalized estimated equations were used for comparisons between groups after adjusting for inter-eye associations. Of 394 eyes of 200 patients, 223 (57%) had ßPPA. The ßPPA eyes were older (59.6 ± 13.7 vs. 56.5 ± 13.7 year, P = 0.02), more myopic (-4.0 ± 3.5 vs. -1.3 ± 3.5 D, P < 0.01), and had poorer SAP scores (MD: -4.9 ± 5.2 vs. -2.6 ± 5.2 dB, P < 0.01; PSD: 4.3 ± 2.9 vs. 2.5 ± 3.0 dB, P < 0.01). By univariate analysis, mean latencies were longer in ßPPA eyes (6.1 ± 5.3 vs. 4.0 ± 5.5 ms, P < 0.01). After adjusting for differences in SE, age, and SAP MD, there was no significant difference between the two groups (P = 0.09). ßPPA eyes had lower amplitude log SNR (0.49 ± 0.16 vs. 0.56 ± 0.15, P < 0.01), which lost significance (P = 0.51) after adjusting for MD and PSD. Although eyes with ßPPA had significantly lower amplitudes and prolonged latencies than eyes without ßPPA, these differences were attributable to differences in SAP severity, age, and refractive error. Thus, ßPPA does not appear to be an independent factor affecting mfVEP responses in eyes with GON.

Reliability of a computer-aided manual procedure for segmenting optical coherence tomography scans.

PURPOSE: To assess the within- and between-operator agreement of a computer-aided manual segmentation procedure for frequency-domain optical coherence tomography scans. METHODS: Four individuals (segmenters) used a computer-aided manual procedure to mark the borders defining the layers analyzed in glaucoma studies. After training, they segmented two sets of scans, an Assessment Set and a Test Set. Each set had scans from 10 patients with glaucoma and 10 healthy controls. Based on an analysis of the Assessment Set, a set of guidelines was written. The Test Set was segmented twice with a ≥1 month separation. Various measures were used to compare test and retest (within-segmenter) variability and between-segmenter variability including concordance correlations between layer borders and the mean across scans (n = 20) of the mean of absolute differences between local border locations of individual scans, MEAN{mean( ΔLBL )}. RESULTS: Within-segmenter reliability was good. The mean concordance correlations values for an individual segmenter and a particular border ranged from 0.999 ± 0.000 to 0.978 ± 0.084. The MEAN{mean( ΔLBL )} values ranged from 1.6 to 4.7 µm depending on border and segmenter. Similarly, between-segmenter agreement was good. The mean concordance correlations values for an individual segmenter and a particular border ranged from 0.999 ± 0.001 to 0.992 ± 0.023. The MEAN{mean( ΔLBL )} values ranged from 1.9 to 4.0 µm depending on border and segmenter. The signed and unsigned average positions were considerably smaller than the MEAN{mean( ΔLBL )} values for both within- and between-segmenter comparisons. Measures of within-segmenter variability were only slightly larger than those of between-segmenter variability. CONCLUSIONS: When human segmenters are trained, the within-and between-segmenter reliability of manual border segmentation is quite good. When expressed as a percentage of retinal layer thickness, the results suggest that manual segmentation provides a reliable measure of the thickness of layers typically measured in studies of glaucoma.

Automated layer segmentation of macular OCT images using dual-scale gradient information.

A novel automated boundary segmentation algorithm is proposed for fast and reliable quantification of nine intra-retinal boundaries in optical coherence tomography (OCT) images. The algorithm employs a two-step segmentation schema based on gradient information in dual scales, utilizing local and complementary global gradient information simultaneously. A shortest path search is applied to optimize the edge selection. The segmentation algorithm was validated with independent manual segmentation and a reproducibility study. It demonstrates high accuracy and reproducibility in segmenting normal 3D OCT volumes. The execution time is about 16 seconds per volume (480x512x128 voxels). The algorithm shows potential for quantifying images from diseased retinas as well.

A comparison of retinal nerve fiber layer (RNFL) thickness obtained with frequency and time domain optical coherence tomography (OCT).

To diagnose glaucoma and other diseases of the retinal ganglion cell/ optic nerve, the thickness of the retinal nerve fiber layer (RNFL) is routinely measured with optical coherence tomography. Until recently, these OCT measurements were made almost exclusively with a time domain OCT (tdOCT) machine from a single manufacturer. Recently, a number of OCT machines, based upon an improved frequency domain OCT technology (fdOCT), have appeared. We compared measurements made using a new fdOCT machine to those from the older tdOCT machine. The results were comparable. More importantly, we learned that the key factor determining whether results from different machines will be comparable is the algorithm used to segment RNFL thickness, not the type of OCT.

The Association Between Clinical Features Seen on Fundus Photographs and Glaucomatous Damage Detected on Visual Fields and Optical Coherence Tomography Scans.

PURPOSE: To classify the appearance of the optic disc seen on fundus photographs of healthy subjects and patients with or suspected glaucoma whose diagnosis was based upon visual fields (VFs) and spectral-domain optical coherence tomography (sdOCT) results. PATIENTS AND METHODS: One eye of 100 patients with or suspected glaucoma and 62 healthy subjects were prospectively tested with 24-2 and 10-2 VF and macular and disc sdOCT cube scans. All eyes with or suspected glaucoma had a 24-2 mean deviation better than -6.0 dB and an abnormal appearing disc on stereophotographs. The retinal ganglion cell plus inner plexiform layer (RGC+) from the macular scans and the retinal nerve fiber layer (RNFL) from the macular and disc scans were segmented and converted to probabilities plots. An eye was considered "glaucoma" if the sdOCT probability plots showed an abnormality in a region that corresponded to a defect seen on the 24-2 and/or 10-2 VF total deviation plot. Similarly, an eye was considered "suspect" only if both the sdOCT and VF plots were normal. Healthy subjects (normal VFs and sdOCT) were classified as "controls" and used as reference for comparisons. Glaucoma specialists reviewed the stereophotographs and classified eyes based on the presence of signs suggestive of glaucomatous optic neuropathy. RESULTS: The pattern of clinical signs of glaucomatous optic neuropathy seen on stereophotographs was statistically different between glaucoma (P<0.001) and suspects (P<0.001) vs. controls and explained up to 68% of the total variance of the diagnosis based upon sdOCT and VFs. Vertical cup-to-disc>0.6, focal neuroretinal rim thinning, focal RNFL loss, and violation of the ISNT rule had the best performance to differentiate glaucoma and suspects from controls. Compared with the suspect group, glaucoma eyes (abnormal sdOCT and VF tests) were more likely to have vertical cup-to-disc>0.6 (92% vs. 69%, P=0.003), diffuse rim (53% vs. 9%, P<0.001) and RNFL (61% vs. 26%, P<0.001) thinning, and ß-zone parapapillary atrophy (68% vs. 17%, P<0.001). CONCLUSIONS: Focal and diffuse signs of glaucoma damage seen on stereophotographs often match damage shown on VFs and sdOCT. In addition, damage shown on VFs and sdOCT is often missed during clinical evaluation. Longitudinal studies ought to differentiate focal signs of glaucoma damage seen on stereophotography from false-positives or very early loss.

Evaluation of the Structure-Function Relationship in Glaucoma Using a Novel Method for Estimating the Number of Retinal Ganglion Cells in the Human Retina.

PURPOSE: We developed a simple method for estimating the number of retinal ganglion cells (RGCs) in the human retina using optical coherence tomography (OCT), compared it to a previous approach, and demonstrated its potential for furthering our understanding of the structure-function relationship in glaucoma. METHODS: Swept-source (ss) OCT data and 10-2 visual fields (VFs) were obtained from 43 eyes of 36 healthy controls, and 50 eyes of 50 glaucoma patients and suspects. Using estimates of RGC density from the literature and relatively few assumptions, estimates of the number of RGCs in the macula were obtained based on ssOCT-derived RGC layer thickness measurements. RESULTS: The RGC estimates were in general agreement with previously published values derived from histology, whereas a prior method based on VF sensitivity did not agree as well with histological data and had significantly higher (P = 0.001) and more variable (P < 0.001) RGC estimates than the new method based on ssOCT. However, the RGC estimates of the new approach were not zero for extreme VF losses, suggesting that a residual, non-RGC contribution needs to be added. Finally, the new ssOCT-derived RGC estimates were significantly (P < 0.001 to P = 0.018) related to VF sensitivity (Spearman's ρ = 0.26-0.47), and, in contrast to claims made in prior studies, statistically significant RGC loss did not occur more often than statistically significant visual loss. CONCLUSIONS: The novel method for estimating RGCs yields values that are closer to histological estimates than prior methods, while relying on considerably fewer assumptions. Although the value added for clinical applications is yet to be determined, this approach is useful for assessing the structure-function relationship in glaucoma.

Evaluation of a Method for Estimating Retinal Ganglion Cell Counts Using Visual Fields and Optical Coherence Tomography.

PURPOSE: To evaluate the accuracy and generalizability of a published model that derives estimates of retinal ganglion cell (RGC) counts and relates structural and functional changes due to glaucoma. METHODS: Both the Harwerth et al. nonlinear model (H-NLM) and the Hood and Kardon linear model (HK-LM) were applied to an independent dataset of frequency-domain optical coherence tomography and visual fields, consisting of 48 eyes of 48 healthy controls, 100 eyes of 77 glaucoma patients and suspects, and 18 eyes of 14 nonarteritic anterior ischemic optic neuropathy (ION) patients with severe vision loss. Using the coefficient of determination R2, the models were compared while keeping constant the topographic maps, specifically a map by Garway-Heath et al. and a separate map by Harwerth et al., which relate sensitivity test stimulus locations to corresponding regions around the optic disc. Additionally, simulations were used to evaluate the assumptions of the H-NLM. RESULTS: Although the predictions of the HK-LM with the anatomically-derived Garway-Heath et al. map were reasonably good (R2 = 0.31-0.64), the predictions of the H-NLM were poor (R2 < 0) regardless of the map used. Furthermore, simulations of the H-NLM yielded results that differed substantially from RGC estimates based on histology from human subjects. Finally, the value-added of factors increasing the relative complexity of the H-NLM, such as assumptions regarding age- and stage-dependent corrections to structural measures, was unclear. CONCLUSIONS: Several of the assumptions underlying the H-NLM should be revisited. Studies and models relying on the RGC estimates of the H-NLM should be interpreted with caution.