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Background: Neurodegenerative diseases are public health challenges in aging populations. Early identification of people at risk for neurodegeneration might improve targeted treatment. Noninvasive, inexpensive screening tools are lacking but are of great potential. Optical coherence tomography (OCT) measures the thickness of nerve cell layers in the retina, which is an anatomical extension of the brain and might be indicative of common underlying neurodegeneration. We aimed to determine the association of macular ganglion cell-inner plexiform layer (mGCIPL) thickness with cognitive and sensorineural function in midlife., Method: This cross-sectional study included 1,880 Beaver Dam Offspring Study participants (aged 27-93 years, mean 58) who participated in the 10-year follow-up examination. We assessed cognitive function and impairment, hearing sensitivity thresholds and impairment, central auditory processing, visual impairment, and olfactory impairment. We measured mGCIPL using the Cirrus 5000 HD-OCT Macular Cube Scan. Multivariable linear and logistic regression models adjusted for potential confounders were used to determine associations between mGCIPL thickness and cognitive and sensorineural functions, as well as for comparing participants with a thin mGCIPL (1 SD below average) to the remainder in those functions., Results: Thinner mGCIPL was associated with worse cognitive function, worse central auditory function, and visual impairment. We found an association of mGCIPL thickness with hearing sensitivity in women only and no association with impairment in hearing, olfaction, and cognition. Results on the thin group comparisons were consistent., Conclusions: mGCIPL thickness is associated with cognitive and sensorineural function and has the potential as a marker for neurodegeneration in middle-aged adults., (© The Author(s) 2020. Published by Oxford University Press on behalf of The Gerontological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Objective: Niemann-Pick disease type C1 (NPC1) is a rare autosomal-recessive lysosomal storage disorder presenting with a broad clinical spectrum ranging from a severe infantile-onset neurovisceral disorder to late-onset neurodegenerative disease. Optical coherence tomography (OCT) is established to detect retinal degeneration in vivo. We examined NPC1-patients (NPC1-P), clinically asymptomatic NPC1-mutation carriers (NPC1-MC), and healthy controls (HC) to (1) identify retinal degeneration in NPC1-disease and (2) to investigate possible subclinical retinal degeneration in NPC1-MC., Methods: Fourteen NPC1-P, 17 NPC1-MC, and 31 age-matched HC were examined using spectral-domain OCT. Neurological examinations, clinical scales [modified Disability Rating Scale (mDRS); Scale for the Rating and Assessment of Ataxia (SARA); Spinocerebellar Ataxia Functional Index (SCAFI)], and video-oculography (VOG) were correlated with OCT data., Results: Macular retinal nerve fiber layer and volumes of combined ganglion cell and inner plexiform layer were significantly lower in NPC1-P compared to HC [mRNFL (µm):0.13 ± 0.01 vs. 0.14 ± 0.02; p = 0.01; GCIPL (mm 3 ):0.60 ± 0.05 vs. 0.62 ± 0.04; p = 0.04]. No significant differences were found in NPC1-MC in comparison to HC. In NPC1-P, the amplitude of upward vertical saccades showed positive associations with peripapillary RNFL (ρ = 0.645; p < 0.05), and thinned GCIP (ρ = 0.609; p < 0.05), but not in NPC1-MC. In NPC1-P correlations between combined outer plexiform layer and outer nuclear layer (OPONL) with mDRS (r = - 0.617; p < 0.05) and GCIP with SARA (r = - 0.622; p < 0.05) were observed. Furthermore, in NPC1-MC, motor scores were negatively associated with pRNFL (ρ = - 0.677; p < 0.01)., Conclusions: Using OCT, we showed retinal degeneration in NPC1-P and significant correlation between retinal neuroaxonal degeneration with clinical measurements. We observed a non-significant trend of retinal degeneration in NPC1-MC correlating with subclinical motor abnormalities. Based on these preliminary data, OCT may be an important marker of neurodegeneration in NPC1-disease after onset of clinical symptoms.

Background/aims: Optical coherence tomography (OCT) is commonly used to diagnose and assess diabetic macular oedema (DME). Swept-source OCT (SS-OCT) promises improved imaging depth and more independence from media opacities. Heidelberg Spectralis full-depth imaging (FDI) combines details at different depths to one representation. The aim of this study was to determine the comparability of the imaging methods concerning DME ultrastructure., Methods: Two graders assessed the presence of typical DME phenomena in eyes with centre-involving DME on Topcon Atlantis SS-OCT and Heidelberg Spectralis FDI spectral-domain OCT (SD-OCT) B-scans. Retinal layer segmentation was corrected and choroidal layers were manually segmented. Graders measured cyst and subretinal fluid (SRF) diameters and counted hyper-reflective foci (HRF). Findings were recorded and statistically analysed., Results: Statistically significant systematic biases (Spectralis-Atlantis) were found for the HRF count (outside the central mm, -6.39, p=0.0338), chorioretinal thickness (central mm: -35.45 µm, p=0.00034), choroidal thickness (central mm: -60.97 µm, p=0.00004) and Sattler's layer thickness (-42.69 µm, p=0.0001). Intergrader agreement was excellent or very good for posterior vitreous detachment, vitreomacular attachment (central mm) and SRF presence in both devices. Manually delineated Sattler's layer thickness showed an intraclass correlation of 0.85 with FDI SD-OCT but 0.26 with SS-OCT (p=0.003)., Conclusion: Prominent aspects such as cysts in the outer nuclear layer and SRF can be identified with comparable confidence, while a significant systematic bias underlies chorioretinal, choroidal and Sattler's layer thickness and HRF count. Specialists should use the same device at every examination during longitudinal clinical consideration or cross-sectional evaluation of these ultrastructural biomarkers., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2020. No commercial re-use. See rights and permissions. Published by BMJ.)

Purpose: To report on the combined use of trypan blue (TB) and brilliant blue G (BBG) for staining the epiretinal membrane (ERM) and internal limiting membrane (ILM) during vitrectomy and to describe the histopathological findings., Methods: 10 surgical specimens were removed from 10 eyes with macular pucker during vitrectomy using a commercially available combination of TB and BBG for ERM and ILM staining and peeling. Specimens were evaluated using light and transmission electron microscopy., Results: In all cases the combination of TB and BBG was useful for identifying and delineating ERM and ILM. No complications related to the use of the dye were observed during or after surgery. Glial cells were present in all specimens. Hyalocytes were observed in 6 cases and myofibroblasts in 3 of them. In 7 cases native vitreous collagen fibrils were found on the ILM, while in 5 specimens newly formed collagen was present. No clinical evidence of toxicity was observed during the 3-month follow-up., Conclusion: The combined use of TB and BBG appeared to be very useful intraoperatively to improve the visualization of ERM and ILM, thus facilitating their complete removal. Anatomical and histopathological findings demonstrated the safety and the efficacy of this vital dye., (© 2018 S. Karger AG, Basel.)

Background: Many optical coherence tomography (OCT) studies have reported alterations in the retinal nerve fiber layer (RNFL) in Parkinson's disease (PD) and other neurodegenerative diseases. However, whether retinal alterations are a biomarker for PD is still controversial., Objective: To investigate potential correlations between PD and morphological changes in retina using OCT and to determine its usefulness as a biomarker of disease progression in PD., Methods: We performed a cross-sectional study on patients with PD (N = 37) and age-matched controls (N = 42), followed by a longitudinal study of the PD patients (N = 22) over approximately 2.5 years., Results: The average retinal nerve fiber layer (RNFL) thickness (p < 0.001), total macular thickness (p = 0.001), and macular volume (p = 0.001) were decreased in PD patients compared to controls and had further decreased at the follow-up visit (p < 0.05 for all). The average RNFL thickness and the total thickness of macular were negatively correlated with age in PD patients at baseline. Linear regression analysis revealed that age (p = 0.002, p = 0.003, respectively) and LEDD (p = 0.011, p = 0.013, respectively) were correlated to total thickness and volume of macular in 22 PD patients in the follow-up study. However, no correlation was found between RNFL and other parameters., Conclusions: PD progression is associated with pronounced retinal structure changes, which can be quantified by OCT. Patterns of RNFL and macular damage detected by the noninvasive technology of OCT can be a useful biomarker for evaluating the progression of PD.

Purpose: The ultrastructural anatomy of the vitreomacular interface in young human donor eyes and animal eyes is explored using scanning electron microscopy (SEM) to determine its relationship with the formation of the perimacular ridge from abusive head trauma, as well as macular hole formation, vitreomacular traction syndrome, and preretinal hemorrhage., Materials and Methods: SEM is used to image the posterior poles of 23 human donor eyes, as well as several cow, dog, monkey, pig, and rabbit eyes for vitreomacular interface anatomy. We examined autopsy eyes from abusive head trauma histopathologically., Results: Two rings of thick, circumferential, vitreous attachment at the area centralis are found. An inner ring at the fovea, R1, and an outer ring at the perifoveal region, R2, are both observed in eyes from donors < 30 years of age; comparatively, in eyes from donors > 30 years, only R2 is present (p<0.001). R2 is found with unique elliptical shape in Cynomolgus monkey. Macula, R1, and R2 are not detected in cow, dog, pig, or rabbit eyes., Conclusions: The vitreomacular ring attachments found in donor eyes correspond anatomically with the perimacular ridge found histopathologically in abusive head trauma, and likely correlates with the macular hole, vitreomacular traction syndrome, and preretinal hemorrhage. Vitreomacular interface anatomy in the monkey, but not the cow, dog, pig, or rabbit, demonstrates some anatomical similarity to that of the human, consistent with species differences regarding the area centralis.

Purpose: The purpose of this study was to identify the structural and histological effects of a Tano diamond-dusted membrane scraper (DDMS) on the retinal surface after internal limiting membrane (ILM) abrasion in macular hole surgery., Methods: Institutional experimental study was performed in 11 eyes. All eyes underwent ILM abrasion in the operating room with a DDMS for macular hole repair as an alternative to traditional ILM peeling. Three human donor eyes underwent an identical procedure in the laboratory. Retinal tissues were removed by ILM abrasion with a DDMS during vitrectomy for macular hole repair and retinal tissues remaining in human donor eyes. Main outcome measures were microscopic and immunohistological characteristics of instrument tip tissues and retinal structure after ILM abrasion., Results: The tips of the Tano DDMS showed evidence of cellular membranes and ILM removal. The retinas showed distinct areas of lamellar ILM removal without penetration of the retinal nerve fiber layer (RNFL)., Conclusions: Application of the Tano DDMS instrument is sufficient to remove membranes from the surface of the ILM and layers of the ILM without disruption of the underlying RNFL. Internal limiting membrane abrasion can be a useful and effective alternative to complete ILM removal for macular surgery.

Mouse models have greatly assisted our understanding of retinal degenerations. However, the mouse retina does not have a macula, leading to the question of whether the mouse is a relevant model for macular degeneration. In the present study, a quantitative comparison between the organization of the central mouse retina and the human macula was made, focusing on some structural characteristics that have been suggested to be important in predisposing the macula to stresses leading to degeneration: photoreceptor density, phagocytic load on the RPE, and the relative thinness of Bruch's membrane. Light and electron microscopy measurements from retinas of two strains of mice, together with published data on human retinas, were used for calculations and subsequent comparisons. As in the human retina, the central region of the mouse retina possesses a higher photoreceptor cell density and a thinner Bruch's membrane than in the periphery; however, the magnitudes of these periphery to center gradients are larger in the human. Of potentially greater relevance is the actual photoreceptor cell density, which is much greater in the mouse central retina than in the human macula, underlying a higher phagocytic load for the mouse RPE. Moreover, at eccentricities that correspond to the peripheral half of the human macula, the rod to cone ratio is similar between mouse and human. Hence, with respect to photoreceptor density and phagocytic load of the RPE, the central mouse retina models at least the more peripheral part of the macula, where macular degeneration is often first evident.

A/J mice develop progressive hearing loss that begins before 1 month of age and is attributed to cochlear hair cell degeneration. Screening tests indicated that this strain also develops early onset vestibular dysfunction and has otoconial deficits. The purpose of this study was to characterize the vestibular dysfunction and macular structural pathology over the lifespan of A/J mice. Vestibular function was measured using linear vestibular evoked potentials (VsEPs). Macular structural pathology was evaluated using light microscopy, scanning electron microscopy, transmission electron microscopy, confocal microscopy and Western blotting. Individually, vestibular functional deficits in mice ranged from mild to profound. On average, A/J mice had significantly reduced vestibular sensitivity (elevated VsEP response thresholds and smaller amplitudes), whereas VsEP onset latency was prolonged compared to age-matched controls (C57BL/6). A limited age-related vestibular functional loss was also present. Structural analysis identified marked age-independent otoconial abnormalities in concert with some stereociliary bundle defects. Macular epithelia were incompletely covered by otoconial membranes with significantly reduced opacity and often contained abnormally large or giant otoconia as well as normal-appearing otoconia. Elevated expression of key otoconins (i.e., otoconin 90, otolin and keratin sulfate proteoglycan) ruled out the possibility of reduced levels contributing to otoconial dysgenesis. The phenotype of A/J was partially replicated in a consomic mouse strain (C57BL/6J-Chr 17(A/J)/NaJ), thus indicating that Chr 17(A/J) contained a trait locus for a new gene variant responsible to some extent for the A/J vestibular phenotype. Quantitative trait locus analysis identified additional epistatic influences associated with chromosomes 1, 4, 9 and X. Results indicate that the A/J phenotype represents a complex trait, and the A/J mouse strain presents a new model for the study of mechanisms underlying otoconial formation and maintenance.

The macula is a site where various vitreoretinal disorders occur. In 1983 we started to observe the retinal surface of postmortem eyes with a scanning electron microscope (SEM). We investigated the anatomy of the vitreous in postmortem eyes by slit lamp biomicroscopy. The novel anatomy of the premacular vitreous led us to conduct a clinical study of vitreomacular interface diseases. In 1997, time domain optical coherence tomography(OCT) became available which facilitated visualization of the vitreoretinal interface. Swept source OCT which was introduced in 2012 can depict liquefied lacunae in the vitreous. It enabled us to elucidate the mechanism of vitreoretinal diseases. I. SEM revealed the remnants of vitreous cortex at fovea with high incidence (44%), which suggests strong vitreoretinal attachment at the fovea and vitreous cortex origin of the epiretinal membrane. II. We studied the anatomy of the vitreous in postmortem eyes. The vitreous of bisected eye balls was stained by fluorescein and immersed in water and observed by slit-lamp biomicroscopy. We discovered a "posterior precortical vitreous pocket (PPVP)" in adult eyes without posterior vitreous detachment (PVD). III. We performed clinical study in various vitreoretinal diseases based on the novel vitreous anatomy and explained their mechanism. 1. In diabetic retinopathy, ring shaped fibrovascular tissue surrounding the macula is formed along the outer margin of the PPVP. Although PVD progresses outside the PPVP, its posterior wall remains attached to the retina, which causes macular traction or cystoid macular edema. 2. In eyes with idiopathic epimacular membrane (IEM), detached vitreous cortex had an oval defect corresponding to the IEM. Posterior wall of the PPVP that is premacular vitreous cortex appeared to be the framework of IEM. 3. During vitrectomy for macular hole, premacular round defect appears when PVD is created. The residual cortex on the macula is fibrous membrane with elasticity. The tangential contraction of premacular cortex may generate anterior traction to the fovea, which leads to macular hole. IV. Using time domain OCT, we demonstrated the evolution of macular hole, myopic foveoschisis and lamellar macular hole. After 2007, we investigated age related changes of vitreoretinal interface by spectral domain OCT V. We demonstrated whole structure of the PPVP using swept source OCT. PPVP is a boat shaped premacular liquefied space which has a connecting channel to Cloquet's canal. PPVP develops during childhood. Visualization of vitreous structure proved that our previous assumptions are reasonable. Although the physiological function of the PPVP is unclear, we speculate that the aqueous flows into the PPVP though Cloquet's canal and the connecting channel.

Triamcinolone acetonide (Kenacort) is a corticosteroid that can be administrated by subconjunctival injection, with an extended release for up to three months. Our retrospective study aims to analyze safety and efficacy of subconjunctival triamcinolone injections in the treatment of uveitic macular edema. We included 31 eyes of 30 patients, who had one or several injections. We studied the progression of visual acuity, central macular thickness by optical coherence tomography (OCT), intraocular pressure, and presence or absence of cataract, on the day of injection (T0), and at 1, 3, 6 and 12 months after injection. Twenty-one patients had only one injection; 10 patients had 2. The 12-month follow-up showed an improvement in visual acuity with an initial mean of 0.36 ± 0.27 logMAR to 0.23 ± 0.33 logMAR at 3 months of follow-up (P<0.0004), and to 0.24 ± 0.21 logMAR at 12 months (P=0.0371), for a two-line improvement. A decrease in mean central macular thickness was measured by OCT, from a mean of 444 ± 112 μm (0.24 ± 0.11 logSD-OCT) at T0 to 355 ± 103 μm (0.14 $ ± 0.10 logSD-OCT) at 3 months (P=0.0002). We did not find a significant increase in intraocular pressure, and we diagnosed one cataract during follow-up but this occurred in the uninjected eye as well. Subconjunctival injection of triamcinolone acetonide is a safe and effective treatment of macular edema related to uveitis. Initial clinical monitoring is necessary to detect iatrogenic events., (Copyright © 2014 Elsevier Masson SAS. All rights reserved.)

Background: Photocoagulation reduces the incidence of visual loss in diabetic patients with focal macular edema, but it can induce it for Efficacy of topical ketorolac for improving visual function after photocoagulation in diabetic patients with focal macular edema 6 weeks after treatment and produces visual improvement in some cases. Topical ketorolac may reduce the inflammation caused by photocoagulation and improve visual outcome., Purpose: To determine the efficacy of topical ketorolac for improving visual function after photocoagulation in diabetic patients with focal macular edema., Methods: An experimental, comparative, prospective, longitudinal study in diabetic patients with focal macular edema was conducted. Eyes were randomized into two groups of topical treatment for 3 weeks after photocoagulation (A: ketorolac, B: placebo). Best corrected visual acuity before and after treatment was compared in each group (paired t test), and the proportion of eyes with visual improvement was compared between groups (χ(2)). The evaluation was repeated after stratifying for initial visual acuity (≥ 0.5, < 0.5)., Results: There were 105 eyes included. In group A (n= 46) mean visual acuity changed from 0.50 to 0.58 (p= 0.003), and from 0.55 to 0.55 in group B (n= 59, p= 0.83); mean percent change was 22.3% in group A and 3.5% in group B (p= 0.03). Visual improvement was identified in 25 eyes from group A (54.3%) and 19 from group B (32.2%, p= 0.019, RR 1.65); the difference only persisted when initial visual acuity was ≥ 0.5 (10 [40%], group A, 5 [14.7%], group B, p= 0.02, RR 2.72)., Conclusion: Topical ketorolac was more effective than placebo to improve best corrected visual acuity in diabetic patients with focal macular edema.

Purpose: To investigate microstructural changes and visual prognosis in myopic subretinal hemorrhages (mSH) without choroidal neovascularization (CNV)., Methods: In this retrospective, observational case series, 13 consecutive eyes with mSH were followed for 6 months. The medical records, fluorescein angiography (FA), and spectral-domain optical coherence tomography (OCT) were reviewed. Fluorescein angiography confirmed the absence of CNV. The baseline and 6-month findings/parameters were investigated, including the maximal hemorrhagic height, intraretinal hyperscattering signal across the retina at 6 months (i.e., intraretinal hyperreflective sign), and integrity of the photoreceptor inner and outer segment (IS/OS) and external limiting membrane (ELM) lines., Results: The final visual acuity (VA) improved significantly (P = 0.001), and the hemorrhages resolved in 12 (92.3%) eyes by 6 months. The tops of the hemorrhages reached the outer nuclear layer (ONL) in three eyes (23.1%), internal limiting membrane (ILM) in five (38.5%), and between the two layers in five (38.5%). The intraretinal hyperreflective sign in all eyes extended into the ONL in five eyes (38.5%), to the ILM in four (30.8%), and between the two layers in four (30.8%). The location of the hyperreflective signs at 6 months coincided with the ruptured retinal layers at baseline in all eyes. The IS/OS line and the ELM were each intact in six (46.2%) eyes. The final VA was associated significantly with the IS/OS (P < 0.05) and ELM (P < 0.01) integrity., Conclusions: The intraretinal hyperreflective sign, presumed to be scarring that enters through the disrupted outer retina, is correlated closely with photoreceptor function.

To investigate whether heavy habitual smoking affects microstructures and functions of the macula, 45 age- (20-39 years old) and sex-matched adult smokers (≥1 box/day for ≥5 years) and 45 nonsmokers (controls) were enrolled in this case-control study. Central macular thickness (CMT), macular autofluorescent pigment density (MAPD), macular electroretinogram (ERG), and photostress recovery time (PRT) measurements were performed. The mean age of smokers and nonsmokers was 32.9 ± 3.9 and 33.1 ± 4.1 years, respectively (p = 0.43), and smoking duration was 11 ± 5.6 years. CMT in smokers (220 ± 28 μm) and nonsmokers (217.2 ± 31 μm; p = 0.57) was similar. Smokers had lower MAPD values (124.6) than nonsmokers (138.2) (p = 0.010). Multifocal ERG parameters in the central (6°) hexagon were similar in both groups (p > 0.05 for latency and amplitudes of P1 and N1). PRT in smokers and nonsmokers was similar (7.2 ± 1.2 and 7.4 ± 1.9 min, respectively; p = 0.33); however, foveal threshold value (FTV) at the first minute after photostress was statistically higher in smokers (36.1 ± 1.04 dB) than nonsmokers (34.8 ± 1.05 dB) (p = 0.011). We conclude that decreased MAPD and altered response to photostress may be indicative of early nicotine toxicity in microstructurally sound macula of adult chronic smokers.

Objective: To establish normal values of macular thickness and volume obtained by the Cirrus SD-OCT (Carl ZeissMeditec, Dublin, CA, U.S.A.). Secondly, to assess the association between macular thickness and volume, sex and age., Material and Methods: A prospective study was conducted on patients who were seen in a hospital Retina Unit, and who only had retinal disease in one eye. All the Macular Cube 512 × 128 scan protocols were performed by the same operator. Only the healthy eye was scanned in each patient., Results: A total of 100 eyes of 100 patients were analysed. The mean central foveal thickness was 261.31 ± 17.67 microns, and was significantly (P<.05) higher in males (267.74 ± 16.98 microns) than in females (255.60 ± 16.40 microns). The mean obtained for the volume of the cube was 10.09 ± 0.37mm 3, and the mean thickness of 280.33 ± 10.34 cube um, with no statistically significant differences between gender being found (P<.05). The mean macular thickness is less at central level, increases in the inner perifoveal ring, and then decreases in the outer perifoveal ring. Furthermore, of all quadrants the greatest thickness was the nasal (328.27 ± 12.96 microns), followed by the upper (326.27 ± 11.89 microns), lower (322.53 ± 12.37mm) sectors, with the temporal sector being the thinnest (313.35 ± 14.20 microns). The mean age of the patients was 60.86 ± 14 years., Conclusion: The mean central foveal thickness and the thickness of the inner perifoveal ring are significantly higher in men than in women. Both the mean volume and thickness of the cube, as well as nasal and inner superior sectors decrease with age, being significantly only in women., (Copyright © 2012 Sociedad Española de Oftalmología. Published by Elsevier Espana. All rights reserved.)

Purpose: To study macular morphology in amblyopic eyes using high-definition spectral domain optical coherence tomography (SD-OCT) and to compare the findings with fellow eyes., Methods: This was a prospective institutional study of patients ≥6 years of age with unilateral amblyopia. Enhanced high-definition single line macular scans of amblyopic eyes were obtained using SD-OCT and compared with fellow eyes. Scans were evaluated qualitatively for structural differences. Central foveal thickness was measured and areas of the different retinal layers were computed within 500 µm from the foveal centre nasally and temporally., Results: Forty-five patients with unilateral amblyopia were included: 25 with strabismic and 20 with anisometropic amblyopia with a mean age of 24.8 years. Qualitatively, the bulge in the inner segment/outer segment junction of the central fovea was noted to be attenuated or absent in 60% of amblyopic eyes compared with 29% of normal eyes, p=0.02. Also, amblyopic eyes demonstrated a shallow foveal pit compared with normal fellow eyes. Mean foveal thickness was significantly increased in amblyopic (228.56 µm) versus fellow eyes (221.72 µm), p=0.03. Upon exploring different retinal layers, the temporal inner nuclear layer area was increased (p=0.04) while the outer nuclear layer area was decreased (p=0.04) in amblyopic eyes compared with fellow eyes., Conclusions: Using enhanced high-definition SD-OCT, amblyopic eyes demonstrated qualitative and quantitative differences in macular features, possibly representing signs of immaturity compared with normal fellow eyes.

Purpose: morphobiometric evaluation of macula and peripapillary retinal nerve fiber layer thickness with the use of high-resolution optical coherence tomography in children with a history of regressed retinopathy or prematurity., Materials and Methods: 18 patients at the age of 8 to 14 years with a history of spontaneously regressed retinopathy of prematurity were studied prospectively. For statistical purposes a control group of 21 matched subjects at the age of 8 to 15 years was used. Ophthalmic examination and optical coherence tomography were performed in each patient. Peripapillary nerve fibre layer thickness, foveal and parafoveal thickness ratio, total macular volume and subfoveal choroidal thickness were measured in both groups., Results: in the optical coherence tomography, the foveal thickness in children with retinopathy of prematurity was significantly higher [269.5 μm (232-321)] compared to the controls [224.5 μm (207-267)]. The macular volume in the study group was also higher (8.68 mm³). The subfoveal choroidal thickness was reduced in study group [321 μm (112-365)] compared to the control group [337 μm (294-358)]. There was no statistical significant difference in total peripapillary nerve fibre layer thickness between the two groups., Conclusion: The morphobiometric macular changes in eyes with a history of regressed retinopathy of prematurity are possibly related to the developmental abnormalities, which retinopathy of prematurity is due to the presence of the abnormal foveal structure across all retinal layers.

This article summarizes the results of the BRAVO and CRUISE trials, two randomized multicenter studies in patients with macular edema secondary to branch and central retinal vein occlusion, respectively. Randomization was 1:1:1 to 0.3 mg of ranibizumab, 0.5 mg of ranibizumab or placebo. Monthly injections were administered for 6 months followed by a 6-month observation period in which treatment on an on-demand (PRN) basis was applied with 0.5 mg ranibizumab. Patients in the control group were also eligible for 0.5 mg ranibizumab treatment in the observation period. The results showed a significant anatomical and visual improvement in both treatment groups 7 days after the intravitreal injection. PRN treatment with monthly follow-up maintained the visual improvements achieved after the first 6 months of treatment. Patients in the control group who received PRN treatment after the first 6 months showed an anatomical improvement similar to that in the treatment groups but less visual improvement.

Background: Optical coherence tomography (OCT) quantifies changes of foveal thickness and macular volume after photocoagulation in diabetic macular edema. Macular volume evaluates the whole macula, but it may underestimate changes in foveal thickness induced by photocoagulation. We undertook this study to evaluate the concordance between macular volume and foveal thickness for identifying clinically significant changes of retinal thickness after photocoagulation for diabetic macular edema., Methods: We carried out an observational, retrospective, longitudinal, analytical study. Center point thickness (CPT), central subfield mean thickness (CSMT) and macular volume were measured with OCT before photocoagulation and 3 weeks after in diabetic patients with focal macular edema (January 2006--January 2010). Concordance among variables to detect clinically significant changes (CPT >17%, CSMT >11%, macular volume >3%) was identified using the kappa test., Results: Sixty eight eyes were included; 47 eyes had nonproliferative retinopathy (69.1%). CPT increased significantly in 14.7% of the sample; CSMT in 8.8%, and macular volume in 11.8%. CPT decreased significantly in 4.4%, CSMT in 8.8%, and macular volume in 42.6%. Concordance was regular for CPT and CSMT increased (57%). Concordance was good for CPT and CSMT decreased (64%). Concordance was regular for CSMT and macular volume decreased in eyes with center involvement (43%). The remaining concordances were poor., Discussion: Two independent events happen after focal photocoagulation: involution of the original thickening and increase in CPT. In order to detect both events, evaluation of either foveal thickness alone or macular volume alone is insufficient., Conclusion: Identifying the efficacy and safety of treatments for diabetic macular edema requires simultaneous measurement of CPT and macular volume.

Purpose: To correlate the clinical and histopathologic features of Best vitelliform macular dystrophy (BVMD)., Methods: Two eyes were obtained postmortem from a patient with BVMD. The patient's clinical information was reviewed. Series sections of the globes were performed and sequentially stained with hematoxylin-eosin, periodic acid-Schiff or Masson trichrome. A section of the left macula was submitted for electron microscopic processing. Histopathologic findings were reconstructed in a scaled two-dimensional map and compared with fundus photography, fundus autofluorescence (FAF), fundus fluorescein angiography (FFA) and optical coherence tomography (OCT) images., Results: The macular lesion of the right eye was identified as a well-demarcated region with pigment, elevated submacular yellow material and subretinal fluid. This corresponded histopathologically to a well-circumscribed area of RPE hyperplasia, accumulation of lipofuscin in the RPE, deposition of granular material in the photoreceptors, macrophages and drusen. The left eye displayed a 1 disc diameter chorioretinal scar with surrounding shallow fluid and submacular pigment. This corresponded to RPE changes and a fibrocellular proliferation in the choriocapillaris., Conclusion: Histopathologic mapping revealed retinal edema, RPE abnormalities, drusen and a chorioretinal scar in BVMD that correlated with the fundus, FFA, FAF and OCT findings.

Purpose: To examine structural differences in the retinal pigmented epithelium (RPE) and Bruch's membrane of rhesus monkeys (Macaca mulatta) as a function of topography and age., Methods: The retinas of two old (24 and 26 years old) and two young (1 and 6 years old) female monkeys were examined by light fluorescence and electron microscopy at the macula, equator, and ora serrata., Results: All monkeys lacked fluorescence and lipofuscin granules in the RPE at the ora serrata where photoreceptors are absent. The equator and macula showed intense fluorescence and many lipofuscin granules in the RPE of the old but not the young monkeys. At the ora, the RPE contained many dense round melanin granules throughout the cell. At the equator and macula, melanin granules were more apical, less frequent, and often elongated. Mitochondria were clustered at the basal side of the RPE cell near infolds of the plasma membrane. Both mitochondria and infolds tended to increase toward the macula. In all regions, the basal lamina of the RPE did not penetrate the extracellular space adjacent to infolds. The elastin layer of Bruch's membrane was wide at the ora and equator and thinner at the macula. In the old monkeys, drusen were found at all retinal regions between the basal lamina and the internal collagen layer of Bruch's membrane. The drusen were often membrane-bound with a basal lamina and contained material resembling structures in the RPE., Conclusions: Lack of fluorescence and lipofuscin in the RPE at the ora serrata, where photoreceptors are absent, confirms that RPE fluorescence occurs only where outer segments are phagocytized. Mitochondrial clustering indicates that the basal side of the RPE cell uses the most energy and this becomes maximal at the macula. The presence of age-related degenerative changes and drusen at all retinal locations in the older monkeys, even at the ora where RPE lipofuscin was absent, indicates that these processes are not dependent on local lipofuscin accumulation. Therefore lipofuscin toxicity may not be the sole cause of age-related RPE degeneration.

Background/aims: To evaluate the ultrastructural changes in the macula after successful repair of rhegmatogenous retinal detachment (RRD) using simultaneous spectral domain optical coherence tomography (sdOCT) and fundus autofluorescence (FAF) imaging and to perform functional correlations with microperimetry., Methods: Simultaneous sdOCT and FAF imaging were performed using a combined sdOCT-confocal scanning laser ophthalmoscope (cSLO) system. Microperimetry was performed in a subgroup of these patients., Results: 17 patients with macular-on RRD and 20 with macular-off RRD were recruited. sdOCT images revealed disruption of the external limiting membrane, inner segment/outer segment junction or the Verhoeff membrane in 16 eyes (43.2%). The presence of any or more of these disruptions was significantly associated with a poor postoperative best-corrected visual acuity (BCVA) (p<0.001). Patients with abnormal FAF were also associated with poor postoperative BCVA (p<0.001). However, there was a generally poor correspondence between the localisation of ultrastructural abnormalities as detected by sdOCT and FAF changes. Areas of decreased microperimetric sensitivity corresponded well with the ultrastructural changes and abnormal FAF., Conclusions: FAF changes may lag behind ultrastructural changes in the OCT, some of which may reverse with the passage of time. The present study demonstrates the ultrastructural changes assessed with simultaneous FAF and sdOCT and their relation with visual outcome.

Purpose: To evaluate the histologic features of cellular retinal fragments on the internal limiting membrane (ILM) removed during idiopathic macular epiretinal membrane (MEM) peeling surgery with and without the aid of indocyanine green (ICG) diluted in 5% glucose., Methods: ILM specimens removed from 88 eyes during idiopathic MEM surgery between 1995 and 2003 were reviewed retrospectively. Histologic analysis focused on the presence and characteristics of retinal fragments on the retinal surface of the ILM. Statistical analysis compared the results between group I (conventional surgery) and group II (ICG-assisted peeling)., Results: Seventy-one eyes underwent MEM surgery without the aid of ICG (group I) and seventeen underwent MEM ICG-assisted surgery (group II). The amount of Müller cell debris on the retinal surface of the ILM was more significant in the group I than in the group II (40.8 vs. 11.8; P = 0.024). Large fragments of Müller cells were more frequently observed in the group I (no ICG) than in the group II (ICG) (63.4 vs. 23.5%; P = 0.003)., Conclusions: The use of ICG diluted with 5% glucose in ILM removal during MEM surgery was associated with less retinal debris attached to the retinal face of the ILM compared with surgery in which ICG was not used.

We demonstrate histologically sub-retinal drusenoid debris in three aged human eyes, two of them affected by age-related maculopathy. By postmortem fundus examination, the lesions were drusen-like, i.e., they were pale spots apparently at the level of the retinal pigment epithelium (RPE). Light and electron microscopy revealed aggregations of membranous debris, the principal constituent of soft drusen, in the sub-retinal space. Immunohistochemistry and confocal microscopy confirmed the presence of molecules typically associated with drusen (positive for unesterified cholesterol, apoE, complement factor H, and vitronectin) without evidence for molecules associated with photoreceptors (lectin-binding disaccharide bridges and opsins), Müller cells (glial fibrillary acid protein and cellular retinal binding protein, CRALPB), or RPE (CRALPB). The fact that a drusenoid material, sharing some markers with conventional drusen, can occur on opposite faces of the RPE, suggests deranged polarity of normally highly vectorial processes for basolateral secretion from RPE, and that overproduction of secreted materials and direction of secretion are independently specified processes. In the future, drusenoid sub-retinal debris might be more frequently revealed by emerging high-resolution imaging techniques.

Purpose: To examine the ultrastructural correlates of spectral-domain optical coherence tomography (SD-OCT) findings in patients with vitreomacular traction (VMT)., Design: Observational case series., Methods: Retrospective analysis of six eyes of consecutive patients who underwent vitrectomy surgery for VMT was performed in this single-center, noncomparative study. One patient had a concurrent macular hole. Preoperative assessment included SD-OCT examination with 3-dimensional image reconstruction. During surgery the vitreous cone was dissected from the vitreous body using scissors, then removed from the surface of the retina with a combination of sharp dissection and peeling, and subsequently submitted for histologic and transmission electron microscopic processing., Results: SD-OCT showed prominent vitreal-foveal adhesion in all six eyes. Each eye had an epiretinal membrane (ERM) under the detached perifoveal posterior vitreous detachment. In all eyes this ERM appeared to course up the cone of attached vitreous and along the back surface of the posterior vitreous face. Ultrastructural analysis showed fibrocellular proliferations in the vitreous specimens in all six cases, which included retinal pigment epithelium (RPE) cells (five eyes), fibrocytes (four eyes), and macrophages (three eyes)., Conclusions: The adhesion between the vitreous and fovea in vitreomacular traction syndrome is accompanied by fibrocellular proliferation along the exposed surfaces of the inner retina and the posterior surface of the vitreous. This fibrocellular proliferation may augment the adhesion between the vitreous and fovea, and may account for the prominent OCT signal seen along the posterior surface of the vitreous in these cases.

Purpose: To determine the size and regional distribution of lipoprotein-like particles (LLPs) that accumulate with age in Bruch's membrane (BrM)., Methods: The quick-freeze/deep-etch method was used to prepare specimens of human BrM (age range, 27-78) for electron microscopic examination. Stereologic methods were used to analyze the resultant micrographs and determine the age-related changes of the LLP volume fraction and diameter distribution in various locations in BrM., Results: The volume fraction occupied by LLPs was found to increase monotonically with age in both the inner collagenous layer (ICL) and elastic layer (EL), but not in the outer collagenous layer (OCL). The mass of total LLP-associated lipids in BrM also increased with age. There was no significant increase in LLP size with age, but there was a modest increase in size with increased volume fraction of LLPs in BrM., Conclusions: The pattern of accumulation of particles was consistent with a retinal pigment epithelium (RPE) source for the LLPs, which explains why once the EL and ICL were filled with particles, LLPs continued to accumulate near the RPE, but no further accumulation was found in the OCL. The quantity of LLP-associated lipids found in BrM accounts for a large portion of the accumulated lipids measured in this tissue.

We compared the ultrastructure of the inner limiting membrane (ILM) and epiretinal tissue in closed and non-closed, idiopathic macular holes (MH). Peeling of ILM and epimacular tissue during vitrectomy was successfully performed on 77 eyes with stage III MH and on 19 eyes with stage IV MH. In 16 additional eyes with non-closed MH, we performed a second vitrectomy with extended ILM removal. The specimens were processed for transmission electron microscopy. Fibrocellular proliferation at the vitreal side of the ILM was found in 57% of MH that were closed by one operation, and in 100% of non-closed MH. Mono- and multilayered cellular membranes as well as native vitreous collagen at the ILM were significantly more frequent in eyes with stage IV MH than in eyes with stage III MH. In non-closed MH, cellular proliferation was mostly seen as irregular cell accumulation, and masses of newly formed collagen were found. Since ILM remnants and collagen represent a stimulus for the early formation of tangential traction preventing successful MH closure, it appears mandatory to create a complete vitreoretinal separation and to remove the ILM and collagen thoroughly during MH surgery.

Deposits in macular human Bruch's membrane (BrM) increase with age and have been postulated to be associated with age-related maculopathy. We used two ultrastructural methods to compare these deposits by electron microscopy in macular and peripheral BrM of eight eyes from donors 63-86 years of age. Quick-freeze/deep-etch (QFDE) was used to prepare replicas that showed the ultrastructure of deposits, and osmium-tannic acid-paraphenylenediamine (OTAP) was used to preserve small extracellular lipid particles. We found that an accumulation of lipoprotein-like particles (LLPs) occurred in the peripheral BrM just as it does in the macular region, but with perhaps a somewhat slower time course. The "lipid wall," reported in macular BrM, was also found occasionally in the peripheral regions. The same processes that lead to age-related accumulation of LLPs in macular BrM appear to also occur in the peripheral regions.

Lipid-containing inclusions have been observed in human Bruch's membrane (BrM) and are postulated to be associated with age-related maculopathy (ARM), a major cause of legal blindness in developed countries. The dehydration associated with specimen preparation for thin-section transmission electron microscopy causes loss of these inclusions. Better preservation of the ultrastructure of the inclusions and tissue is achieved by using a quick-freeze/deep-etch preparation. We use this technique to examine normal human macular BrM in order to characterize the deposition of the lipid-rich inclusions and their age-related accumulation within different layers of the tissue. We find that various inclusions mentioned in other studies can be formed by combinations of three basic structures: lipoprotein-like particles (LLPs), small granules (SGs) and membrane-like structures. These inclusions are associated with collagen and elastic fibrils by fine filaments. In younger eyes, these inclusions are found mostly in the elastic (EL) and outer collageneous layer (OCL) and occupy a small fraction of the interfibrillar spacing. As age increases, LLPs and SGs gradually fill the interfibrillar spacing of the EL and inner collageneous layer (ICL) of the tissue, and later form a new sublayer, the lipid wall, within the boundary region between the basal lamina of retinal pigment epithelium (RPE) and ICL. Because the formation of the lipid wall only occurs after these inclusions fill the ICL, and it seems unlikely that the LLPs can pass through the packed layer, this result suggests a possible RPE origin of the LLPs that make up the lipid wall.

Age-related macular degeneration (AMD) is characterized by progressive loss of central vision, which is attributed to abnormal accumulation of macular deposits called "drusen" at the interface between the basal surface of the retinal pigment epithelium (RPE) and Bruch's membrane. In the most severe cases, drusen deposits are accompanied by the growth of new blood vessels that breach the RPE layer and invade photoreceptors. In this study, we hypothesized that RPE secreted proteins are responsible for drusen formation and choroidal neovascularization. We used stable isotope labeling by amino acids in cell culture (SILAC) in combination with LC-MS/MS analysis and ZoomQuant quantification to assess differential protein secretion by RPE cell cultures prepared from human autopsy eyes of AMD donors (diagnosed by histological examinations of the macula and genotyped for the Y402H-complement factor H variant) and age-matched healthy control donors. In general, RPE cells were found to secrete a variety of extracellular matrix proteins, complement factors, and protease inhibitors that have been reported to be major constituents of drusen (hallmark deposits in AMD). Interestingly, RPE cells from AMD donors secreted 2 to 3-fold more galectin 3 binding protein, fibronectin, clusterin, matrix metalloproteinase-2 and pigment epithelium derived factor than RPE cells from age-matched healthy donors. Conversely, secreted protein acidic and rich in cysteine (SPARC) was found to be down regulated by 2-fold in AMD RPE cells versus healthy RPE cells. Ingenuity pathway analysis grouped these differentially secreted proteins into two groups; those involved in tissue development and angiogenesis and those involved in complement regulation and protein aggregation such as clusterin. Overall, these data strongly suggest that RPE cells are involved in the biogenesis of drusen and the pathology of AMD.

Aim: To demonstrate the ultrastructure of vitreomacular traction associated with persistent hyaloid artery., Methods: Pars plana vitrectomy was performed in a 66-year-old man with progressive vitreomacular traction associated with a persistent hyaloid artery. Epimacular tissue was peeled and processed for transmission electron microscopy., Results: Ultrastructural analysis revealed multiple sheets of cellular and collagenous components. Myofibroblasts and newly formed collagen were the predominant features. Fibrous astrocytes, fibroblasts, macrophages, and basement membrane were also present., Conclusion: The cellular composition of the epimacular tissue and high cellular activity suggest that persistence of the hyaloid artery may contribute to the development of vitreomacular traction.

Age-related macular degeneration (AMD) is a leading cause of blindness in the elderly. In its severest form, choroidal neovessels breach the macular Bruch's membrane, an extracellular matrix compartment comprised of elastin and collagen laminae, and grow into the retina. We sought to determine whether structural properties of the elastic lamina (EL) correspond to the region of the macula that is predilected toward degeneration in AMD. Morphometric assessment of the macular and extramacular regions of 121 human donor eyes, with and without AMD, revealed a statistically significant difference in both the integrity (P < 0.0001) and thickness (P < 0.0001) of the EL between the macular and extramacular regions in donors of all ages. The EL was three to six times thinner and two to five times less abundant in the macula than in the periphery. The integrity of the macular EL was significantly lower in donors with early-stage AMD (P = 0.028), active choroidal neovascularization (P = 0.020), and disciform scars (P = 0.003), as compared to unaffected, age-matched controls. EL thickness was significantly lower only in individuals with disciform scars (P = 0.008). The largest gaps in macular EL integrity were significantly larger in all categories of AMD (each P < 0.0001), as compared to controls. EL integrity, thickness, and gap length in donors with geographic atrophy did not differ from those of controls. These structural properties of the macular EL correspond spatially to the distribution of macular lesions associated with AMD and may help to explain why the macula is more susceptible to degenerative events that occur in this disease.

Background: Recent data on macular hole formation have changed our understanding of the mechanisms involved. However, the mechanisms of macular hole closure, despite the introduction of internal limiting membrane (ILM) removal, are not fully understood. We reviewed the charts of patients who underwent surgery for macular hole with and without intentional removal of ILM and reviewed the basic science literature on the components and relationships of the ILM-glial cell interface to help interpret our surgical data., Methods: We reviewed the records of 205 eyes with a clinical diagnosis of macular hole at a university centre in Montreal between 1998 and 2002. Of these, 117 eyes of 108 patients were included. Vitrectomy with intentional ILM removal was done under indocyanine green (ICG) staining. Macular hole stage and rates of hole closure, failure and reopening were recorded, comparing the eyes with intentional ILM peeling versus no ILM peeling. Anatomic success was defined as a flat/closed configuration. A basic science literature review was done through the PubMed search engine with the key words "internal limiting membrane," "internal limiting lamina," "macular hole," "retinal glial cells" and "retinal Müller cells.", Results: Eighty-four eyes had ICG-guided ILM removal (26 stage 2 macular holes, 45 stage 3 holes and 13 stage 4 holes), and 33 eyes had no ILM removal (10 stage 2 holes and 23 stage 3 holes). Of the 117 eyes 110 (94.0%) were successfully closed. Four (3.6%) of the closed holes reopened (none had ILM peeling), and 7 (6.0%) of the 117 eyes had primary failure. Ten of the 11 eyes that had primary failure or hole reopening were reoperated, and 8 were successfully closed after the second procedure. One patient refused reoperation. Including second procedures, the final anatomic closure rate was 97.4% (114/117)., Interpretation: Intentional ILM peeling is not mandatory for all macular holes to close but does play a role in preventing reopening. Removal of ILM increases the rate of flat/closed hole configuration on reoperation without the need for biologic adjuvants. Review of the basic science evidence helped clarify the possible role of ILM removal.

Objective: To investigate the 3D structure of the macula flava in the human vocal fold., Material and Methods: The 3D structure of the macula flava in the human vocal fold was investigated by means of scanning electron microscopy using a chemical digestion method (modified sodium hydroxide maceration method) and the results compared with those obtained using light microscopy., Results: The macula flava of the human vocal fold comprised a dense mass of cells and extracellular matrices. Many cells were stellate in shape and possessed cytoplasmic processes. Extracellular matrices were composed of collagenous, reticular and elastic fibers, and ground substances (hyaluronic acid). Many fibers ran in various directions. The stellate cells were surrounded by these extracellular matrices. Amorphous materials were attached to the surface of the stellate cells. Extracellular matrices in the macula flava extended to those in the lamina propria of the vocal fold mucosa., Conclusion: Anterior and posterior maculae flavae in the human vocal fold mucosa are postulated to be involved in the metabolism of extracellular matrices, which are essential to the viscoelasticity of the lamina propria (the vibrating tissue) of the human adult vocal fold mucosa.

Purpose: For macular translocation surgery, the native attached retina has to be detached either locally or completely. Although different surgical techniques are used, there is a general search for supporting procedures that facilitate and accelerate the retinal detachment., Methods: Pars plana vitrectomies were performed in pigmented rabbits. In all experimental groups, a local retinal detachment was created by infusing the test solution with a thin glass micropipette attached to a glass syringe. In control animals a standard balanced salt solution was used at room temperature, in combination with a standard vitrectomy light source. In two test groups, a calcium- and magnesium-free solution was used for the vitrectomy, under illumination by a standard light source in group I (solution at room temperature) and group II (solution heated up to body temperature). In group III the rabbits were dark-adapted for half an hour, and then, during surgery, a red filter was used in front of the light source (standard balanced salt solution at room temperature). After the rabbits were killed at the end of surgery, the adherence of the retinal pigment epithelium (RPE) to the neural retina in the detachment area was quantified microscopically, and the morphologic integrity of the detached retinal tissue was examined by light and electron microscopy. No electrophysiology was performed., Results: In all four groups, it was possible to detach the retina. The maximum adherence of the RPE cells to the neural retina was observed in the control group. Virtually no decrease in adherence was found in test group II (36 degrees C solution without calcium and magnesium), whereas a significant decrease was seen in both group I (calcium- and magnesium-free solution at room temperature) and group III (dark adaptation-red light technique; standard balanced salt solution at room temperature). In none of the experimental groups was any obvious damage of the retinal structure observed, even after exposure to the test solutions for 60 minutes., Conclusions: Both dark adaptation (red illumination) and the use of a calcium chloride- and magnesium chloride-free solution (at room temperature) can facilitate retinal detachment in macular translocation surgery. Both techniques are proposed as a gentle support for the operation, because they protect an intact RPE cell layer and do not cause retinal damage at the ultrastructural level.

Aim: To analyze the ultrastructural features of epimacular tissue removed during idiopathic macular hole (IMH) surgery and to compare them with those of idiopathic epimacular membranes., Methods: Three consecutive patients with unilateral stage 3 IMH underwent pars plana vitrectomy with surgical removal of delicate gelatinous membranous epimacular tissue. One patient was previously treated with laser photocoagulation. The excised material was investigated by means of light and transmission electron microscopy. Results were compared with the findings of the excised material in two idiopathic epimacular membranes., Results: The excised material included variable proportions of cellular and membranous material. Glial cells (Müller cells, fibrous astrocytes) and fibroblast-like cells dominated in the cellular component both in the specimens from IHM eyes and in the comparative material. Neural elements were revealed in two of three specimens from IMH eyes. Membraneous material was formed by fragments of internal limiting membrane., Conclusions: Dominance of the glial cells in the cellular component of epimacular formations may reflect the important role of these cells both in formation and in healing process of IMH. The presence of neural elements in the removed epimacular tissue suggests that, at least in some cases, IMH surgery with peeling of an epimacular membrane can be associated with some injury of the macula.

Purpose: Vitreomacular traction syndrome is characterized by an incomplete posterior vitreous detachment and by the cortical vitreous remaining attached to the macula. In certain case epimacular membrane is clinically well defined. We studied the electron microscopic features of epiretinal tissue and posterior hyaloid removed from the posterior pole., Material and Methods: [corrected] Six epimacular membranes were removed during a pars plana vitrectomy, placed in 2.5% glutaraldehyde solution, embedded in Epoxy resin, and examined in a transmission electron microscope. The detached posterior hyaloid was removed and analyzed separately in two cases., Results: The analysis disclosed five morphologically cell types. Fibrous astrocytes were predominant in five cases. Other cell types were less frequent and were identified as fibroblasts, macrophages and myofibroblasts. In one case epithelial-like cell type was predominant. The stroma was mainly composed of a thin layer of 15-nm diameter collagen evoking a vitreous origin. Internal limiting membrane was present in one case only. Detached posterior hyaloid was constituted by the same cellular constituents on thin layer of vitreous collagen., Discussion: The cellular proliferation is principally constituted by fibrous astrocyte and occurs on the inner surface of cortical vitreous. Some glial cells seem to present an epithelial differentiation., Conclusion: The attached and detached posterior hyaloid in vitreomacular traction syndrome is the place of glial cells migration and proliferation.

Purpose: To test the hypothesis that neovascular age-related macular degeneration is related to oxidative stress involving the macular retinal pigment epithelium. This study investigated, as a function of age, levels of enzymes that defend tissues against oxidative stress in the macular retinal pigment epithelium of human eyes with this disease., Methods: Surgical specimens of macular choroidal neovascular membranes from eyes with age-related macular degeneration and the macular regions of whole donor eyes with neovascular age-related macular degeneration or without evident ocular disease were studied by quantitative electron microscopic immunocytochemistry with colloidal gold-labeled second antibodies. Relative levels in retinal pigment epithelium cell cytoplasm and lysosomes were determined of five enzymes believed to protect cells from oxidative stress, as well as levels of the retinal pigment epithelium marker cytoplasmic retinaldehyde-binding protein, for comparison with the enzymes., Results: Copper, zinc superoxide dismutase immunoreactivity increased and catalase immunoreactivity decreased with age in cytoplasm and lysosomes from macular retinal pigment epithelium cells of normal eyes and eyes with age-related macular degeneration. Cytoplasmic retinaldehyde-binding protein immunoreactivity showed no significant relationship to age or the presence of neovascular age-related macular degeneration. Glutathione peroxidase immunoreactivity was absent from human retinal pigment epithelium cells. Both heme oxygenase-1 and heme oxygenase-2 had highly significantly greater immunoreactivity in retinal pigment epithelium cell lysosomes than in cytoplasm, differing from the much greater cytoplasmic immunoreactivity of the other proteins studied. This immunoreactivity decreased with age, particularly in the lysosomes of retinal pigment epithelium cells from eyes with neovascular age-related macular degeneration. These decreases were of borderline significance (P = .067 for heme oxygenase-1; P = .12 for heme oxygenase-2) when eyes with age-related macular degeneration were compared with normal eyes by multivariable logistic regression., Conclusions: The high heme oxygenase-1 and heme oxygenase-2 lysosomal antigen levels in macular retinal pigment epithelium cells of eyes with neovascular age-related macular degeneration suggest that oxidative stress causes a pathologic upregulation of these enzymes. Increased lysosomal disposal may indicate that the reparative functions of these enzymes are accompanied by deleterious effects, necessitating their rapid removal from the cell. The much higher heme oxygenase-1 and heme oxygenase-2 antigen levels in macular retinal pigment epithelium cells from younger individuals suggest that protective mechanisms against oxidation and, hence, presumably to the development of age-related macular degeneration, decrease with age.

The purpose of this study was to assess surgical results of internal limiting membrane (ILM) maculorrhexis in macular hole surgery. This study is a part of continuing prospective clinical trial of our team of researchers. Thirteen eyes of 13 patients with idiopathic macular hole underwent vitrectomy with the removal of posterior cortical vitreous, peeling of the macular ILM, and intraocular gas tamponade, followed by postoperative face-down positioning. The excised specimens were evaluated with transmission electron microscopy. Complete closure of the hole was observed in all 13 eyes (100% anatomic success rate). Visual improvement of 2 or more lines on ETDRS visual acuity chart was achieved in 11 (85%) of the 13 eyes. Six (54.5%) eyes attained visual acuity of 20/50 or better. Electron microscopy showed ILM in the removed specimens. ILM maculorrhexis is a promising new surgical approach to close idiopathic macular holes but requires further investigation and long-term evaluation.

Efficacy and possibility of direct operations on the macular area of the retina in humid (exudative) maculopathies (senile maculodystrophy) is validated theoretically, morphologically, and clinically. A hypothesis on the mechanism of exudative process development in the macular area in degenerative diseases is formulated on the basis of pathohistological analysis of removed subpigmental material. Indications for such operations are defined.

Purpose: To present the clinicopathologic features of the eyes of a patient with age-related macular degeneration (ARMD): the right eye was treated for well-defined extrafoveal choroidal neovascularization (CNV), and the left eye had an untreated disciform scar., Methods: The patient was studied ophthalmoscopically and by fluorescein angiography at the time of presentation and on follow-up examinations up to 54 days after laser treatment, when he died. The posterior portions of both eyes (obtained postmortem), including the macula and optic nerve head, were sectioned serially for light microscopy. Tissue sections from both eyes were removed from glass slides and studied by transmission electron microscopy., Results: Histopathologic study of the right eye disclosed a thin layer of basal laminar deposit throughout the posterior pole. Two defects in Bruch's membrane without CNV were present within the area of laser photocoagulation located superior to the fovea. No CNV was present in the scar. Eleven areas of early CNV were present in the posterior pole. Histopathologic study of the left eye showed a prominent basal laminar deposit throughout the posterior pole. A 2.6 x 2.7 mm disciform scar was present that was located mostly in the subretinal space. Four sources of CNV were present., Conclusions: The clinicopathologic features of a treated eye with well-defined extrafoveal CNV, and the fellow eye with a disciform scar, both associated with ARMD, are presented. Although laser treatment obliterated a choroidal neovascular membrane, 11 additional areas of early, subclinical CNV were present.

Purpose: To illustrate the histologic changes at the macula of an eye shielded from focused light for 76 years. Age-related macular degeneration with a central, subretinal, fibrous scar was present in the fellow eye., Methods: Autopsy examination of both eyes of a 91-year-old man was performed. The superior half of the macular area of the shielded eye was studied by light microscopy, and the inferior half was studied by electron microscopy. Serial sections through the macular area of the fellow eye were prepared and studied., Results: The shielded eye contained basal laminar deposits up to 6.0 microns in thickness and basal linear deposits up to 1.0 micron in thickness in the macular area. The photoreceptor outer and inner segments of the outer nuclear layer were intact. The macular area of the fellow eye contained a two-component, vascularized, disciform scar that measured up to 250 microns in one area, with retinal pigment epithelial tears at the nasal and temporal margins. Extensive photoreceptor layer atrophy was present., Conclusions: Such marked asymmetry of macular degeneration in this case may be further evidence of the role of light exposure in the development of age related macular degeneration.

Modern surgery of the macula exists since 1965, and was initially proposed in membrane removal from patients presenting an idiopathic epiretinal membrane. This first indication authorized the development of macular microsurgical instrumentation and further analysis of membrane fragments. However, the exact pathogenesis of these membranes remains yet unknown. Since 1991 macular surgery was applied in the treatment of macular holes. Results obtained after macular hole surgery confirmed pathogenic hypothesis observed clinically by Gas. Considerable improvement was therefore achieved in anatomical and functional outcome. Chorioretinal adhesion agents were recently proposed to optimize the closure of macular holes. Submacular surgery is the latest technologic achievement under investigation. The ultimate indications are age-related macular degeneration, first cause of blindness in industrialized countries. Retinal transplantation opens new avenues in the treatment of a large number of chorioretinal diseases. The assessment of macular surgery could pave the way in realising such technical advances.

Immunogold cytochemistry was used to investigate the fine structural distribution of collagen types I-VI in Bruch's membrane and choroid of the aged human macula. Macular tissue was obtained from ten eyes, and processed for cryoultramicrotomy and London Resin white embedding. Striated collagen fibrils within the inner and outer collagenous layers were found to contain collagen types I, III and V. In addition, type V collagen was also present in the basement membrane of the choriocapillaris. Gross thickening of the choriocapillaris basement membrane was attributed to the deposition of type IV collagen. However, type IV collagen appeared to be absent from the basement membrane of the retinal pigment epithelium. The interesting location of type VI collagen on the choroidal side of the choriocapillaris suggested that its function is to anchor the choriocapillaris onto the choroid. The collagens studied were absent from fibrous banded material, long-spacing collagen, the elastic layer and amorphous granular material. It was concluded that, of the collagen types studied, only the deposition of type IV collagen contributes to the age-related thickening of Bruch's membrane.

The formation of basal laminar deposit (BLD) is one of the histopathologic changes in the aging human macula. BLD is assumed to be an early stage of age-related macular degeneration. The location of BLD, between the RPE plasma membrane and its basement membrane and in the outer collagenous zone of Bruch's membrane, and its ultrastructure suggest that it is composed of excessive amounts of basement membrane material. The main components of basement membranes are type IV collagen, heparan sulfate proteoglycans (HSPG) and laminin. Labeled antibodies against these components can therefore be used for the identification and localization of basement membrane material by means of immunohistochemical techniques. In this study the presence of type IV collagen, laminin and HSPG was determined in aged human maculae by immunohistochemistry and immunoelectron microscopy. Tests for the presence of type VI collagen and fibronectin were also performed. We obtained 76 eyes from 68 human subjects at autopsy or after surgical enucleation for anteriorly located choroidal melanomas. The finely granular component of BLD stained positive with antibodies against type IV collagen, HSPG and laminin, but the long-spacing collagen component of BLD did not. Neither component of BLD was stained with antibodies against type VI collagen or fibronectin. We conclude that BLD consists partly of excess basement membrane material.

In subretinal neovascularisation capillaries originating from the choriocapillaris must cross Bruch's membrane to reach the subretinal pigment epithelial space. Thus gaps in Bruch's membrane have to be formed before subretinal neovascularisation. Histological examination of eyes with subretinal neovascularisation or disciform scars has shown macrophages adjacent to thin areas and ruptures in Bruch's membrane. This has been interpreted as phagocytosis of Bruch's membrane. The purpose of this study was to investigate whether immune complex depositions can be detected in maculae with early stages of age-related macular degeneration and to explain the macrophage reaction before the disciform reaction. A series of 20 human maculae were examined by direct immunofluorescence light microscopy to detect the presence of immune complexes with antibodies directed against immunoglobulins, fibrinogen, and complement factors. Transmission electron microscopy on several maculae was performed to identify the macrophages. Macrophages were observed in close relation to the readily recognisable long spacing collagen, which suggested that long spacing collagen was selectively internalised by these cells. Definite immune complex depositions were not found in basal laminar deposits or drusen. Linear deposits of fibrinogen and complement were frequently found in the outer collagenous zone of Bruch's membrane. However, because of the absence of immunoglobulins, it seems unlikely that these non-specific deposits might cause chemoattraction of macrophages and play a role in the initial phase of the development of subretinal neovascularisation and disciform macular degeneration.

A basal laminar deposit (BLD) in the human macula has been described as an early sign of age-related macular degeneration. In some eyes with a BLD in the macula, light microscopic sections of the peripheral retina revealed almost similar deposits between the retinal pigment epithelium and Bruch's membrane. Because the exact pathogenesis of age-related macular degeneration and the origin of the BLD are unknown, we studied the ultrastructure of these peripheral sub-RPE deposits. Parts of the equatorial and peripheral regions of the retina of ten human eyes, with BLD-like deposits between the retinal pigment epithelium and Bruch's membrane, were examined by electron microscopy. In eight of these ten eyes the ultrastructure of these deposits was amorphous and finely granular. Five of the eight deposits also contained small amounts of long-spacing collagen. Ultrastructurally, the deposits were similar to an early type BLD in the macula. In the remaining two eyes, the deposits appeared to consist of flat, elongated drusen. Our findings indicate that a BLD can develop not only in the macula but also in the peripheral region of the retina.

Scleral tissue from the region of the human macula was studied by the immunogold labeling technique (cryoultramicrotomy and LR white resin embedding) in an attempt to identify the fine structural distribution of collagen types I-VI. Labeling of the striated collagen fibrils suggested colocalisation of collagen types I, III and V with type V occurring at the fibril surface. Both types V and VI collagen were localised to filamentous strands in the interfibrillar matrix. Collagen types II and IV were absent from the scleral stroma.