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Purpose: Given the similarities between the retinal and coronary microvasculature, the retina holds promising potential to serve as a non-invasive screening tool for coronary artery disease (CAD). We aimed to develop novel inner ellipse-based metrics and discern whether foveal avascular zone (FAZ) alterations can serve as indicators for CAD presence and severity., Methods: Patients admitted to the Department of Cardiology who underwent coronary angiography were included. This resulted in an inclusion of 212 patients, of which 73 had no CAD. During the same visit, 6 × 6-mm (nominal size) fovea-centered optical coherence tomography angiography images of both eyes were acquired. The Gensini score (GS) was utilized to quantify CAD severity. Six known FAZ shape metrics were assessed and three novel biomarkers based on the inner ellipse were defined: absolute inner ellipse difference, Hausdorff distance, and Chamfer distance., Results: Eight out of nine metrics showed significant associations with the GS in the left eye. However, significant differences across three CAD severity groups were only demonstrated by the novel metrics. Utilizing the Chamfer distance, age, and sex, patients with and without CAD could be distinguished with an average area under the curve (AUC) of 0.89 (95% confidence interval [CI], 0.84-0.95). Moreover, three CAD severity groups could be discerned with a macro average AUC of 0.77 (95% CI, 0.72-0.84)., Conclusions: A comprehensive assessment of FAZ shape descriptors was performed, and a strong association with CAD was found. The inner ellipse-based biomarkers especially demonstrated high predictive abilities for CAD presence and severity.

Purpose: To investigate if changes in vessel density (VD) and the foveal avascular zone (FAZ) occur in the preclinical phase of Alzheimer's disease (pAD) over time., Methods: Optical coherence tomography angiography (OCTA) was used to image VD and FAZ at baseline and for a follow-up period of 2 years. Positron emission tomography (PET) was used to determine the amyloid beta (Aβ) status of participants., Results: The VD and FAZ of 148 participants (54% female) were analyzed at baseline and follow-up (mean time between measurements, 2.24 ± 0.35 years). The mean age of the participants was 68.3 ± 6.0 years at baseline and 70.3 ± 5.9 years at follow-up. Participants were divided into three groups: control group, participants who had negative Aβ status at both measurements (Aβ-, n = 116); converter group, participants who transitioned from negative to positive between baseline and follow-up (Aβ-+, n = 18); and participants who were consistently positive at both visits (Aβ++, n = 14). The VD of both Aβ+ groups demonstrated non-significant increases over time in both macula and optic nerve head (ONH) regions. The Aβ- group was found to be significantly higher in both ONH and macular regions. The VD of the Aβ++ group was significantly higher in the macula inner and outer rings compared to the Aβ-+ and Aβ- groups. No significant change was found in FAZ values over time., Conclusions: Alterations in VD seem to manifest already in pAD, exhibiting distinct variations between the ONH and macula. Further longitudinal studies with a longer follow-up design and known amyloid pathology should be undertaken to validate these observations.

Purpose: The purpose of this study was to identify and measure plexus-specific absolute retinal capillary blood flow velocity and acceleration in vivo in both nonhuman primates (NHPs) and humans using erythrocyte mediated angiography (EMA) and optical coherence tomography angiography (OCTA)., Methods: EMA and OCTA scans centered on the fovea were obtained in 2 NHPs and 11 human subjects. Scans were also obtained in NHP eyes while IOP was experimentally elevated. Erythrocyte velocity and acceleration in retinal arteries, capillaries, and veins were measured and capillaries were categorized based on location within the superficial vascular (SVP), intermediate capillary (ICP), or deep capillary plexus (DCP). Generalized linear mixed models were used to estimate the effects of intraocular pressure (IOP) on capillary blood flow., Results: Capillary erythrocyte velocity at baseline IOP was 0.64 ± 0.29 mm/s in NHPs (range of 0.14 to 1.85 mm/s) and 1.55 ± 0.65 mm/s in humans (range of 0.46 to 4.50 mm/s). Mean erythrocyte velocity in the SVP, ICP, and DCP in NHPs was 0.69 ± 0.29 mm/s, 0.53 ± 0.22 mm/s, and 0.63 ± 0.27 mm/s, respectively (P = 0.14 for NHP-1 and P = 0.28 for NHP-2). Mean erythrocyte velocity in the human subjects did not differ significantly among SVP, ICP, and DCP (1.46 ± 0.59 mm/s, 1.58 ± 0.55 mm/s, and 1.59 ± 0.79 mm/s, P = 0.36). In NHPs, every 1 mm Hg increase in IOP was associated with a 0.13 mm/s reduction in arterial velocity, 0.10 mm/s reduction in venous velocity, and 0.01 mm/s reduction in capillary velocity (P < 0.001) when accounting for differences in mean arterial pressure (MAP)., Conclusions: Blood flow by direct visualization of individual erythrocytes can be quantified within capillary plexuses. Capillary velocity decreased with experimental IOP elevation.

Purpose: In aging and early-intermediate age-related macular degeneration (AMD), rod-mediated dark adaptation (RMDA) slows more at 5° superior than at 12°. Using optical coherence tomography angiography (OCTA), we asked whether choriocapillaris flow deficits are related to distance from the fovea., Methods: Persons ≥60 years stratified for AMD via the Age-Related Eye Disease Study's nine-step system underwent RMDA testing. Two adjacent 4.4° × 4.4° choriocapillaris OCTA slabs were centered on the fovea and 12° superior. Flow signal deficits (FD%) in concentric arcs (outer radii in mm, 0.5, 1.5, 2.2, 4.0, and 5.0 superior) were correlated with rod intercept time (RIT) and best-corrected visual acuity (BCVA)., Results: In 366 eyes (170 normal, 111 early AMD, 85 intermediate AMD), FD% was significantly worse with greater AMD severity in all regions (overall P < 0.05) and poorest under the fovea (P < 0.0001). In pairwise comparisons, FD% worsened with greater AMD severity (P < 0.05) at distances <2.2 mm. At greater distances, eyes with intermediate, but not early AMD differed from normal eyes. Foveal FD% was more strongly associated with longer RIT at 5° (r = 0.52) than RIT at 12° (r = 0.39) and BCVA (r = 0.21; all P < 0.0001). Choroidal thickness was weakly associated with longer RIT at 5° and 12° (r = 0.10-0.20, P < 0.05) and not associated with AMD severity., Conclusions: Reduced transport across the choriocapillaris-Bruch's membrane-retinal pigment epithelium complex, which contributes to drusen formation under the macula lutea (and fovea), may also reduce retinoid resupply to rods encircling the high-risk area. FD% has potential as a functionally validated imaging biomarker for AMD emergence.

Purpose: To evaluate microvascular intereye differences in diabetic patients with same-stage diabetic retinopathy (DR) in both eyes as assessed using optical coherence tomography angiography (OCTA)., Methods: In this cross-sectional study, fovea-centered swept-source 6 × 6 mm OCTA scans were acquired using a 200 kHz OCTA device. Vessel density (VD) and fractal dimension were calculated on binarized, vessel-segmented images in the superficial capillary plexus (SCP) and deep capillary plexus (DCP). Foveal avascular zone (FAZ) area (FAZA) and perimeter (FAZP) was measured and FAZ circularity (FAZC) calculated. Absolute difference (δabs) and asymmetry index between eyes was assessed and compared across DR stages. Differences of VD, FD, and FAZ parameters between left and right eye were evaluated using linear mixed models., Results: A total of 336 eyes of 168 diabetic patients without DR and with DR stages ranging from mild nonproliferative to proliferative DR were included for analysis. The intereye comparison revealed significantly lower VD in the SCP (estimate [95% CI] = -0.009 [-0.01; -0.006], P < 0.01), as well as a significantly lower FD in the SCP (-0.007 [-0.009; -0.005], P < 0.01) of the left compared to the right eye. FAZC of the left compared to the right eye was lower in eyes without DR, moderate DR, and PDR (P < 0.05). FAZ δabs and asymmetry index were higher in more advanced disease stages (P < 0.05)., Conclusions: OCTA metrics provide important information on the retinal microvasculature in systemic diseases such as DR. Our results reveal a significant intereye difference with lower VD and FD in the SCP as well as higher FAZ impairment of the left compared to the right eye.

Purpose: To develop Choroidalyzer, an open-source, end-to-end pipeline for segmenting the choroid region, vessels, and fovea, and deriving choroidal thickness, area, and vascular index., Methods: We used 5600 OCT B-scans (233 subjects, six systemic disease cohorts, three device types, two manufacturers). To generate region and vessel ground-truths, we used state-of-the-art automatic methods following manual correction of inaccurate segmentations, with foveal positions manually annotated. We trained a U-Net deep learning model to detect the region, vessels, and fovea to calculate choroid thickness, area, and vascular index in a fovea-centered region of interest. We analyzed segmentation agreement (AUC, Dice) and choroid metrics agreement (Pearson, Spearman, mean absolute error [MAE]) in internal and external test sets. We compared Choroidalyzer to two manual graders on a small subset of external test images and examined cases of high error., Results: Choroidalyzer took 0.299 seconds per image on a standard laptop and achieved excellent region (Dice: internal 0.9789, external 0.9749), very good vessel segmentation performance (Dice: internal 0.8817, external 0.8703), and excellent fovea location prediction (MAE: internal 3.9 pixels, external 3.4 pixels). For thickness, area, and vascular index, Pearson correlations were 0.9754, 0.9815, and 0.8285 (internal)/0.9831, 0.9779, 0.7948 (external), respectively (all P < 0.0001). Choroidalyzer's agreement with graders was comparable to the intergrader agreement across all metrics., Conclusions: Choroidalyzer is an open-source, end-to-end pipeline that accurately segments the choroid and reliably extracts thickness, area, and vascular index. Especially choroidal vessel segmentation is a difficult and subjective task, and fully automatic methods like Choroidalyzer could provide objectivity and standardization.

Purpose: To evaluate changes in the retinal microvasculature of young adults over 24 hours using optical coherence tomography angiography (OCT-A)., Methods: Participants (n = 44, mean age 23.2 ± 4.1 years, 24 myopes and 20 nonmyopes) with normal ophthalmological findings were recruited. Two macular OCT-A and OCT scans, systemic blood pressure, intraocular pressure (IOP), and biometry measurements were taken every four hours over 24 hours. Superficial and deep retinal layer en face images were analyzed to extract magnification-corrected vascular indexes using image analysis including foveal avascular zone metrics, vessel density, and perfusion density for the foveal, parafoveal, and perifoveal zones., Results: Significant diurnal variations (P < 0.001) were observed in the vessel and perfusion density in the three superficial retinal layer regions, with acrophase between 4:30 PM and 8:30 PM. Only foveal and parafoveal regions of the deep retinal layer exhibited significant diurnal variations with acrophase between 9 AM and 3 PM. Myopes and nonmyopes had different acrophases but not amplitudes in the parafoveal perfusion density of superficial retinal layer (P = 0.039). Significant correlations were observed between diurnal amplitudes or acrophases of superficial retinal layer indexes and systemic pulse pressure, IOP, axial length and retinal thickness., Conclusions: This study shows, for the first time, that significant diurnal variation exists in OCT-A indexes of macular superficial and deep retinal layer over 24 hours and were related to variations in various ocular and systemic measurements. Myopes and nonmyopes showed differences in the timing but not in amplitude of the superficial retinal layer parafoveal perfusion density variations but not in deep retinal layer.

Purpose: The purpose of this study was to evaluate the interocular differences in choroidal vasculature, choriocapillaris perfusion, and retinal microvascular network, and to explore their associations with interocular asymmetry in axial lengths (ALs) in children with anisomyopia., Methods: Refractive error, AL, and other biometric parameters were measured in 70 children with anisomyopia. Using optical coherence tomography (OCT) and OCT-angiography, we measured the submacular choroidal thickness (ChT), total choroidal area (TCA), luminal area (LA), stromal area (SA), choroidal vascularity index (CVI), choriocapillaris flow deficit (CcFD), retinal vessel density (VD), and foveal avascular zone (FAZ) area., Results: The mean interocular differences in spherical equivalent refraction and AL were -2.26 ± 0.94 diopters and 0.95 ± 0.46 mm, respectively. Submacular ChT, TCA, LA, SA, and CVI were all significantly lower in the more myopic (longer AL) eyes than in the less myopic (shorter AL) fellow eyes. In eyes with longer ALs, both the CcFD and FAZ areas were significantly greater, whereas the superficial and deep retinal VDs were significantly less. After adjusting for corneal power and intraocular pressure, interocular differences in LA (β = -0.774), SA (β = -0.991), and CcFD (β = 0.040) were significantly associated with interocular asymmetry in AL (all P < 0.05)., Conclusions: In pediatric anisomyopes, eyes with longer ALs tended to have lower choroidal vascularity and choriocapillaris perfusion than the contralateral eyes with shorter ALs. Longitudinal investigations would be useful follow-ups to test for a causal role of choroidal circulation in human myopia.

Purpose: To evaluate anatomic-functional associations at sites of retinal lesions in retinal vein occlusion (RVO)., Methods: This pilot, prospective, observational study was conducted at the Northern Ireland Clinical Research Facility (NICRF) of Queen's University and the Belfast Health and Social Care Trust, Northern Ireland, between August 1, 2018, and September 30, 2019. The study included 10 treatment-naïve patients with RVO (10 RVO eyes and 10 fellow eyes). There were 81 points/sites assessed for each eye at baseline; six patients were re-assessed 6 months after anti-vascular endothelial growth factor therapy at the same locations. We investigated associations between retinal sensitivity and presence of structural RVO lesions, including retinal ischemia, hemorrhages, intraretinal fluid (IRF) and subretinal fluid outside the foveal/parafoveal regions. Comparisons were made between RVO eyes and fellow eyes at baseline, and between RVO eyes at baseline and at 6 months after treatment. Regression models were used to investigate anatomic-functional associations., Results: At baseline, strong associations were found between reduced retinal sensitivity and presence of ischemia (estimate = -2.08 dB; P < 0.001), intraretinal fluid (estimate = -7.82 dB; P < 0.001), and subretinal fluid (estimate = -8.66 dB; P < 0.001). Resolution of subretinal fluid but not intraretinal fluid was associated with improved function (estimate = 2.40 dB [P = 0.022]; estimate = 1.16 dB [P = 0.228], respectively). However, reperfusion of ischemic retina, observed in 31 of 486 points (6%) 6 months after anti-vascular endothelial growth factor therapy, was associated with a further decrease in retinal sensitivity (estimate = -2.34 dB; P = 0.035)., Conclusions: Retinal sensitivity was decreased at sites of RVO lesions. Decreased function at sites of retinal ischemia did not recover after treatment, even when reperfusion occurred.

Purpose: To compare quantitative optical coherence tomography angiography (OCT-A) measurements of the parafoveal microvasculature in retinal capillary plexuses among Behҫet uveitis (BU) patients, non-ocular Behҫet's disease (NOBD) patients, and healthy volunteers (HVs)., Methods: Sixty-eight subjects were enrolled in this prospective observational cross-sectional study. OCT-A imaging was performed using the Heidelberg Engineering Spectralis OCT. A custom algorithm was developed to calculate the vessel density (VD) in three retinal vascular layers: deep capillary plexus, intermediate capillary plexus, and superficial vascular plexus. The foveal avascular zone (FAZ) and acircularity index were calculated for the whole retinal vascular complex., Results: We analyzed one eye from 21 BU patients (age, 51 ± 10 years), 23 NOBD patients (age, 48 ± 14 years), and 22 HVs (age, 44 ± 13 years). One-way multivariate analysis of covariance showed a statistically significant difference in VD among the three groups when combining the layers after controlling for scan quality (P < 0.001). The VD was lowest in the BU group and highest in the HV group in all layers. The FAZ area was also statistically significant different among the groups (P < 0.005), with the largest FAZ areas in BU patients and smallest FAZ areas in the HV group. However, no statistically significant difference was found for the acircularity index., Conclusions: The parafoveal microvasculature is affected not only in BU patients but also in NOBD patients. Most deviations in the retinal microcirculation in Behҫet patients were found in the deeper layers of the retina by using the quantitative VD measurement.

Purpose: The purpose of this study was to determine systemic stressors, including fasting plasma glucose (FPG), and other major atherosclerotic cardiovascular disease (ASCVD) risk factors of the retinal microvasculature in people without diabetes., Methods: The Kailuan Eye Study enrolled applicants from the community-based longitudinal Kailuan Study. Applicants underwent optical coherence tomographic angiography (OCTA) and systemic examinations. Both the macula and optic disc were screened, whereas superficial capillary plexus (SCP), deep capillary plexus (DCP), foveal vessel density in the 300 µm ring (FD-300), and radial peripapillary capillaries (RPCs) density were measured in the study., Results: This study included 353 eligible applicants (mean age = 49.86 ± 11.41 years; 47% men; FPG =5.32 ± 1.19 mmol/L). Lower DCP density was associated with elder age (P = 0.001), male gender (P < 0.001), and higher FPG (P = 0.008). Male gender (P < 0.001), axial length (P < 0.001), and FPG (P = 0.029) were inversely associated with RPC density. Meanwhile, a higher FPG concentration was significantly correlated with lower DCP density (P = 0.006) and higher intraocular pressure (P = 0.006), after adjusting mean arterial blood pressure (P = 0.001) and sex (P = 0.042)., Conclusions: DCP density showed a significantly negative correlation with FPG concentration in people without diabetes. These data suggest hyperglycemia could cause early retinal capillary alterations in patients without clinical signs of retinopathy and indicate the potential clinical applications of routine OCTA may be beneficial to screen for subclinical microvasculature and monitor patients with high risks of ASCVD.

Purpose: To examine the foveal avascular zone (FAZ) in patients with congenital achromatopsia (ACHM)., Methods: Forty-two patients with genetically confirmed ACHM were imaged either with Optovue's AngioVue system or Zeiss's Plex Elite 9000, and the presence or absence of a FAZ was determined. For images where a FAZ was present and could be confidently segmented, FAZ area, circularity index, and roundness were measured and compared with previously published normative values. Structural optical coherence tomography images were acquired to assess the degree of foveal hypoplasia (number and thickness of inner retinal layers present at the fovea)., Results: A FAZ was present in 31 of 42 patients imaged (74%), although no determination could be made for 11 patients due to poor image quality (26%). The mean ± SD FAZ area for the ACHM retina was 0.281 ± 0.112 mm2, which was not significantly different from the previously published normative values (P = 0.94). However, their FAZs had decreased circularity (P < 0.0001) and decreased roundness (P < 0.0001) compared to the normative cohort. In the patients with ACHM examined here, the FAZ area decreased as the number and thickness of the retained inner retinal layers increased., Conclusions: Our data demonstrate that despite the presence of foveal hypoplasia, patients with ACHM can have a FAZ. This is distinct from other conditions associated with foveal hypoplasia, which generally show an absence of the FAZ. In ACHM, FAZ formation does not appear to be sufficient for complete pit formation, contrary to some models of foveal development.

Purpose: To use structural criteria to reconcile the three-dimensional organization and connectivity of the parafoveal microvasculature., Methods: The parafoveal microvasculature was perfused and labeled in 16 normal human donor eyes for lectin, alpha smooth muscle actin, and filamentous actin. Established structural criteria gathered using confocal microscopy, including vessel diameter, endothelial cell morphology, and presence/density of smooth muscle cells, were used to differentiate arteries, arterioles, capillaries, venules, and veins. Three-dimensional visualization strategies were used to define the connections between retinal arteries and veins within the superficial vascular plexus (SVP), intermediate capillary plexus (ICP), and deep capillary plexus (DCP)., Results: The parafoveal microvasculature has two different inflow patterns and seven different outflow patterns. The SVP and ICP were connected to retinal arteries by arterioles. Inflow into the DCP occurred only via small arterioles (a1; mean diameter, 8.3 µm) that originated from the ICP. Direct connections between the DCP and retinal arteries were not identified. Each capillary plexus formed its own venule that drained independently or in conjunction with venules from other plexuses into a retinal vein at the level of the ganglion cell layer. For the DCP, a1 was significantly smaller than its draining venule (mean diameter, 18.8 µm; P < 0.001)., Conclusions: The SVP and ICP of the parafoveal microvasculature have both in series and in parallel arterial and venous connections. Arterial supply to the DCP originates from the ICP, but with direct drainage to the retinal vein. These findings may help to develop an understanding of the pattern of retinal lesions characterizing a myriad of retinal vascular diseases.

Purpose: The purpose of this study was to compare perfusion parameters of the parafovea with scans outside the parafovea to find an area most susceptible to changes secondary to diabetic retinopathy (DR)., Methods: Patients with different DR severity levels as well as controls were included in this cross-sectional clinical trial. Seven standardized 3 × 3 mm areas were recorded with Swept Source Optical Coherence Tomography Angiography: one centered on the fovea, three were temporal to the fovea, and three nasally to the optic disc. The capillary perfusion density (PD) of the superficial capillary complex (SCC) and deep capillary complex (DCC) as well as the fractal dimension (FD) were generated. Statistical analyses were done with R software., Results: One hundred ninety-two eyes (33 controls, 51 no-DR, 41 mild DR, 37 moderate/severe DR, and 30 proliferative DR), of which 105 patients with diabetes and 25 healthy controls were included (59 ± 15 years; 62 women). Mean PD of the DCC was significantly less in patients without DR (parafovea = 0.48 ± 0.03; temporal = 0.48 ± 0.02; and nasal = 0.48 ± 0.03) compared to controls (parafovea = 0.49 ± 0.02; temporal = 0.50 ± 0.02; and nasal = 0.50 ± 0.03). With increasing DR severity, PD and FD of the SCC and DCC further decreased., Conclusions: Capillary perfusion of the retina is affected early by diabetes. PD of the DCC was significantly reduced in patients with diabetes who did not have any clinical signs of DR. The capillary network outside the parafovea was more susceptible to capillary perfusion deficits compared to the capillaries close to the fovea., Trial Registration: clinicaltrial.gov, NCT03765112, https://clinicaltrials.gov/ct2/show/NCT03765112?term=NCT03765112&rank=1.

Purpose: To investigate the clinical significance of the changes in the macular microvasculature in patients with diabetes mellitus type 2 without diabetic retinopathy., Methods: Fifty-five patients with diabetes mellitus type 2 without diabetic retinopathy and 48 healthy individuals were enrolled in a prospective cross-sectional study. We identified the changes of optical coherence tomography angiography parameters (foveal avascular zone [FAZ] area and circularity, vessel density, and perfusion index) of the 6 × 6-mm macular scan. Correlation and multiple regression analyses were performed between optical coherence tomography angiography parameters and previously known diabetes mellitus type 2-related demographic and systemic characteristics, and serum biochemical markers., Results: FAZ parameters and perfusion index of the superficial and deep vascular plexus showed significant correlation with serum insulin level, and homeostasis model assessment indices. In multiple linear regression analysis, low insulin levels predicted increased FAZ areas in both the superficial (β = -0.007; P = 0.030) and deep layers (β = -0.010; P = 0.018) and a decreased perfusion index in the deep layer (β = 0.003; P = 0.001)., Conclusions: The expansion and loss of circularity of the FAZ and the decrease in the perfusion index may be affected by insulin resistance and secretory capacity in patients with diabetes mellitus type 2 with no diabetic retinopathy.

Purpose: To objectively quantify cone density (CD) and microvascular density (MVD) in normal subjects and patients with moderate or severe retinitis pigmentosa (RP) by adaptive optics (AO) and optical coherence tomography angiography (OCTA) and to evaluate the changes in the parafoveal regions., Method: Thirty-seven eyes from 20 RP patients and 54 eyes from 29 age-matched healthy participants underwent AO fundus and OCTA imaging. AO images covering a 3-mm-diameter area centered on the fovea were subdivided into 5 equidistant concentric rings (C1-C5). An automated algorithm was used to quantify the mean cone density (mCD; cells/mm2). Macular MVDs (%) in the superficial capillary plexus (SCP) and deep capillary plexus (DCP) were assessed by OCTA., Results: In the moderate RP group, CDs in C2 and C3 were each significantly lower than in the normal group (both P < 0.05). In the severe RP group, CDs were significantly lower than in normal eyes in each concentric ring (all P < 0.001; C1-C5). In both RP groups, MVDs were significantly lower than in normal eyes for both the SCP and DCP (both P < 0.05). The mean CD was significantly correlated with the MVD in the DCP (r = 0.43; P = 0.028) but not in the SCP (r = -0.19, P = 0.323)., Conclusions: Decreased CD was present in the moderate and severe RP groups. This was accompanied by a decreased MVD in the DCP. Direct assessment of photoreceptors in RP patients by high-resolution imaging technologies is crucial for the future development of RP therapeutics.

Purpose: To investigate foveal avascular zone (FAZ) changes in the superficial (SCP) and deep (DCP) capillary plexuses in beta-thalassemia major (BTM) patients, as shown in optical coherence tomography angiography., Methods: Nonrandomized, comparative case series of 54 eyes of 27 BTM patients and 46 eyes of 23 healthy controls, utilizing an automated FAZ detection algorithm. Measurements included FAZ area and FAZ shape descriptors (convexity, circularity, and contour temperature). Results were compared between the two groups, and correlated to iron load and chelation therapy parameters., Results: SCP and DCP FAZ area were not significantly different between the control and BTM groups (P = 0.778 and P = 0.408, respectively). The same was true regarding SCP FAZ convexity (P = 0.946), circularity (P = 0.838), and contour temperature (P = 0.907). In contrast, a statistically significant difference was detected between controls and BTM group regarding DCP FAZ convexity (P = 0.013), circularity (P = 0.010), and contour temperature (P = 0.014). Desferrioxamine dosage was strongly correlated to the DCP area (r = 0.650, P = 0.05) and liver magnetic resonance imaging/T2-star to DCP circularity (r = -0.492, P = 0.038). Correlations were also revealed between urine Fe excretion and DCP convexity (r = 0.531, P = 0.019), circularity (r = 0.661, P = 0.002), and contour temperature (r = -0.591, P = 0.008)., Conclusions: Retinal capillary plexuses and especially DCP seem to present unique morphologic changes in BTM patients, not in the FAZ area, but in specific shape descriptors, indicating minor but detectable FAZ changes. These changes correlate well with iron load and chelation therapy parameters. Their clinical importance and pathophysiologic implications remain to be elucidated through further studies.

Purpose: To evaluate vascular changes in the superficial and deep retinal capillary plexus (SCP, DCP) and their association with drusen volume changes in intermediate age-related macular degeneration (iAMD)., Methods: Patients with iAMD were examined at baseline and 12 months thereafter. Drusen volume was extracted from 20° × 20° OCT scans using a 3-mm ETDRS grid using a customized algorithm with manual correction. Vessel density (VD) and flow area (FA) were extracted from 3 × 3 mm SD-OCT-A scans after manual correction of the segmentation. Associations were investigated using multiple regression models., Results: We used 31 eyes of 31 patients for evaluation. The mean age at baseline was 74.9 ± 5.4 years; 26 patients were female. Baseline visual acuity (VA) was 0.05 ± 0.08 logMAR (Snellen equivalent approximately 20/22). The initial mean 3-mm central drusen volume was 0.144 ± 0.136 mm3. A significant association with the signal strength index was consistently found, therefore all capillary measurements were corrected. VD in the same area was 49.88% ± 7.38% and 55.43% ± 9.31% for the SCP and DCP, respectively. The baseline FA resulted in 3.292 ± 0.218 mm2 and 3.433 ± 0.224 mm2 for the SCP and DCP, respectively. No association was found between changes in drusen volume and FA or VD after 12 months (all P > 0.05). VA worsened (P = 0.013) and the foveal FA of the SCP increased significantly (P = 0.014)., Conclusions: No significant association was found between the increase in drusen volume in iAMD and capillary retinal perfusion over a 12-month follow-up. Although VA decreased statistically over this time period, the foveal FA of the SCP increased.

Purpose: To quantify retinal vascular density in amblyopic children and to investigate the relationship between superficial and deep retinal vessel density and retinal thickness., Method: In this prospective cross-sectional study, 85 amblyopic children (5-12 years) and 66 age-matched control subjects participated at a pediatric ophthalmology clinic. Macular vessel densities of the superficial and deep capillary plexuses (SCP, DCP, respectively) and retinal thickness were measured by clinical optical coherence tomographic angiography (OCTA). Vessel density and retinal thickness were compared between amblyopic groups and the control group, and correlations among these two variables were analyzed., Results: Of the 85 amblyopic children, 52 children had anisometropic amblyopia, 16 had strabismic amblyopia, and 17 had bilateral amblyopia. The foveal and parafoveal macular vessel density in the SCP was lower in amblyopic than control children (P ≤ 0.008). Compared to strabismic and bilateral amblyopias, anisometropic amblyopia SCP differed the most from controls (P ≤ 0.005). Macular vessel density in the DCP of amblyopic children was similar to controls. Multiple linear regression analyses showed SCP vessel density was positively correlated with inner retinal thickness in the fovea (P < 0.001) and in the temporal, nasal, and inferior quadrants of the parafovea (P ≤ 0.008)., Conclusions: Macular vessel density is decreased in anisometropic amblyopia, and to a lesser extent, the other amblyopias. Retinal small vessel density was correlated with the thickness of the macular inner retina. The causality of retinal change (i.e., whether it is primary or secondary to the onset of amblyopia) has not yet been determined.

Purpose: To evaluate the microstructure of the fovea in patients with familial exudative vitreoretinopathy (FEVR) compared to healthy controls using optical coherence tomography angiography (OCTA)., Methods: In this consecutive, cross-sectional, observational case series, 41 eyes of 41 patients diagnosed as FEVR and 37 eyes in 37 control subjects were studied. OCTA was utilized to automatically measure the foveal avascular zone (FAZ) and the vessel density (VD). Inner retinal thicknesses (IRT) and central retinal thickness (CRT) were measured with the instrument caliper. Targeted next-generation sequencing was performed, and phenotype-genotype association was analyzed., Results: Small FAZ was found in 31.70% (13/41) FEVR eyes but not in controls. Greater CRT and lower superficial foveal VD were noted in FEVR patients. FAZ is negatively correlated with IRT. Persistence of the inner retinal layer (IRL) in fovea was present in 48.78% (20/41) FEVR eyes but not found in controls. Zero percent (0/10) of patients with the low-density lipoprotein receptor-related protein 5 (LRP5) mutation, 50% (1/2) with the frizzled-4 (FZD4) mutation, and 66.67% (3/4) with the tetraspanin-12 (TSPAN12) mutation had preserved foveal IRL and small FAZ., Conclusions: Our data indicate FEVR status is associated with a significantly smaller FAZ, decreased vascular density in both the superficial and deep layers of parafoveal area, a thicker fovea, and an abnormally preserved IRL in fovea. In addition, patients with the LRP5 mutation had a milder phenotype than those with the FDZ4 or TSPAN12 mutations. These novel findings could provide insight into the understanding of the pathogenesis of FEVR.

Purpose: To assess the relationship between the size of the foveal avascular zone (FAZ; area and perimeter) and vessel density (VD) at various retinal regions using optical coherence tomography angiography (OCTA) in patients with open-angle glaucoma (OAG)., Methods: In this retrospective cross-sectional study, OAG eyes were grouped according to the presence/absence of central visual field (CVF) defects. A control group of normal eyes was also included. OCTA images were obtained from the optic nerve (4.5 × 4.5 mm2) and macular (3 × 3 mm2) area, in which VDs were measured in the circumpapilla, parafovea, and fovea regions. Partial correlation analyses between FAZ parameters representing size and VDs at different retinal regions were performed. The correlation between FAZ parameters and macular ganglion cell-inner plexiform layer thickness (mGCIPLT) and CVF mean sensitivity (MS) was also assessed. For spatial correspondence, correlations between FAZ parameters and corresponding VD, mGCIPLT, and CVF-MS measurements were assessed at each hemiretina., Results: A total of 126 OAG eyes and 40 controls were reviewed. Overall, FAZ parameters showed statistically significant correlations with VDs measured at the fovea, parafovea, and circumpapilla (FAZ area: r = -0.603, -0.385, -0.256; P < 0.001, < 0.001, and 0.003, respectively). These correlations were observed regardless of the presence of CVF defects (all P < 0.05). FAZ size showed significant spatial correlations with VD, mGCIPLT, and CVF-MS measurements at each hemiretina., Conclusions: OCTA-derived FAZ area and perimeter showed statistically significant correlations with VDs measured at various retinal locations with spatial correspondence in OAG.

Purpose: To assess the connection among arterioles, venules, and capillaries in three retinal capillary plexuses using optical coherence tomography angiography (OCTA)., Methods: This was a prospective, cross-sectional, observational study including 20 eyes of 10 healthy subjects. En face and cross-sectional OCTA images were segmented to study the superficial (SCP), middle (MCP), and deep capillary plexuses (DCP). Using thin slabs and manual segmentation within the three plexuses, we examined the connections between the large-caliber superficial vessels within a 3 × 3 mm2 OCTA scan (arterioles and venules) and the smaller capillaries in each plexus., Results: Twenty eyes of 10 healthy subjects (5 females; average age of 30.8 ± 6.3 years) were included in the analysis. We identified vascular interconnections linking the superficial arterioles and venules with capillaries in each plexus (SCP, MCP, and DCP). We found capillaries in the DCP crossed the horizontal raphe., Conclusions: Our findings show that each of the three capillary plexuses in the parafovea has its own feeding arteriolar supply and draining venules, supporting a physiologic model in which each plexus controls its own oxygenated blood supply to match the metabolic needs of each distinct retinal neurovascular unit.

Purpose: To quantify and evaluate macular superficial capillaries and large vessels separately using an optical coherence tomographic angiography (OCTA)-based automatic segmentation algorithm., Methods: In this cross-sectional study, all eyes were scanned using an OCTA device with 3 × 3 mm cube centered on the fovea. Retinal large vessels (arterioles/venules) were automatically segmented from superficial vasculature en-face images. All images were normalized, binarized, and skeletonized for quantification. Metrics of retinal capillaries were calculated by subtracting the measurements of large vessels from total vasculature. Perfusion density (PD), vessel length density (VLD), and vessel diameter index (VDI) within Early Treatment Diabetic Retinopathy Study (ETDRS) 3-mm ring were calculated for total superficial vasculature, large vessels (PDlarge, VLDlarge, and VDIlarge) and capillaries (PDcap, VLDcap, and VDIcap), respectively., Results: Fifty-nine eyes from 59 healthy participants (mean age, 45 ± 14 years, 36 females) and 118 eyes from 67 patients with diabetes mellitus (mean age, 57 ± 10 years, 28 females) were included. The diabetic cohort included four subgroups (35 eyes without diabetic retinopathy, 30 eyes with mild to moderate nonproliferative diabetic retinopathy [NPDR], 27 eyes with severe NPDR, and 26 eyes with PDR). Linear regression showed that all above metrics were correlated with the disease stage (from healthy state to PDR), and the β value was -0.76, 0.24, -0.78, 0.80, 0.30, 0.77, -0.81, 0.16, and -0.82 for VD, VDlarge, VDcap, VDI, VDIlarge, VDIcap, VLD, VLDlarge, and VLDcap, respectively., Conclusions: Retinal capillaries and large vessels responded differently in the context of diabetes. VLD of capillary is a potentially reliable metric in diabetic retinopathy staging.

Purpose: To investigate the relationships between foveal blood flow as measured by laser speckle flowgraphy (LSFG), the retinal-choroidal structure in enhanced depth imaging-optical coherence tomography (EDI-OCT), and central visual function in patients with retinitis pigmentosa (RP)., Methods: We studied 52 consecutive typical RP patients ≤50 years old and 21 age- and sex-matched controls. The mean blur rate (MBR), which represents the blood flow volume, was calculated in a 2.4-mm2 area centered on the fovea by LSFG. Subfoveal horizontal EDI-OCT images were recorded, and the choroidal area, choroidal hyporeflective area, and choroidal hyperreflective area were analyzed in the central 2.4-mm-wide region. The central foveal thickness (CFT), subfoveal choroidal thickness (SCT), and ellipsoid zone (EZ) width were also measured. Visual acuity (VA) and retinal sensitivity (Humphrey 10-2 program) were measured in the RP patients., Results: The MBR, choroidal area, hyporeflective area, hyperreflective area, and SCT were significantly decreased in the RP patients (all P < 0.001, versus controls). Spearman's rank testing demonstrated no significant correlation between the MBR and the choroidal structural parameters in the RP patients. Decreased MBR was significantly associated with reductions in VA, retinal sensitivity, CFT, and EZ width (all P < 0.05). The choroidal structural parameters did not correlate with central visual function, and the choroidal area, hyperreflective area, and SCT were inversely associated with CFT (all P < 0.05)., Conclusions: These results demonstrated the divergence between the choroidal structure and blood function, and suggest that decreased choroidal flow, rather than the structural alteration, is closely associated with foveal degeneration in RP.

Purpose: To identify and quantify the three distinct retinal capillary plexuses and the foveal avascular zone (FAZ) in healthy subjects according to age using optical coherence tomography angiography (OCTA) with novel projection artifact removal (PAR) software and improved segmentation., Methods: All eyes in this cross-sectional study underwent OCTA imaging using RTVue XR Avanti with novel PAR AngioVue software. OCTA scans were analyzed and the three main parafoveal retinal capillary plexuses were segmented and vessel density and FAZ area were calculated., Results: A total of 152 normal eyes from 95 subjects (39 males, 56 females, mean age 42 ± 25 years) were included. The mean vessel density was 15.48 ± 2.04 mm-1 in the superficial retinal capillary plexus (SCP), 15.28 ± 1.82 mm-1 in the intermediate retinal capillary plexus (ICP), and 16.33 ± 2.32 mm-1 in the deep retinal capillary plexus (DCP) for 3 × 3-mm OCTA images. Analysis of 3 × 3-mm scans yielded a mean FAZ area of 0.270 ± 0.101 mm2. The average reduction in vessel density per year of age with 3 × 3-mm OCTA scans was 0.04 mm-1 (0.22%) in the SCP, 0.05 mm-1 (0.27%) in the ICP, and 0.06 mm-1 (0.30%) in the DCP. The average increase in FAZ area per year of age was 0.0015 mm2 (0.72%)., Conclusions: Novel PAR software may provide improved visualization of all three major parafoveal retinal capillary plexuses including the ICP. Using this technology, SCP, ICP, and DCP vessel density decreased with increasing age while FAZ area increased with age.

Purpose: Migraine, particularly with aura, has been associated with ocular and systemic ischemic complications, but there are limited data on the ocular vasculature in migraine. We used optical coherence tomography angiography (OCTA) to assess perfusion of the macula and optic nerve in migraine patients, with (MA) and without (MO) aura, compared to healthy controls (HC)., Methods: We recruited 15 MA (mean age 42 years), 12 MO (mean age 46 years), and 22 HC (mean age 39 years) participants from neurology and neuro-ophthalmology clinics. Participants underwent optical coherence tomography and 3 × 3 mm OCTA of the macula and optic nerve. Foveal avascular zone area was automatically measured using AngioVue software, and vessel density was calculated as blood vessel length divided by scan area (mm-1) after skeletonization of OCTA images., Results: On macular OCTA, MA participants had an enlarged foveal avascular zone area when compared with HC (0.300 ± 0.019 vs. 0.220 ± 0.066 mm2, P = 0.006). In addition, superficial foveal vessel density was decreased in MA participants when compared with MO participants (7.8 ± 0.31 vs. 9.3 ± 0.44, P = 0.04) and HC (7.8 ± 0.31 vs. 9.4 ± 0.21 mm-1, P = 0.002). On optic nerve OCTA, the MA participants had reduced superior peripapillary vessel density when compared with the MO participants (12.0 ± 0.45 vs. 14.0 ± 0.38 mm-1, P = 0.031) and HC (12.0 ± 0.45 vs. 14.1 ± 0.53 mm-1, P = 0.035). There were no significant differences between the MO and HC groups., Conclusions: Migraine with, but not without, aura was associated with foveal and peripapillary vascular decrements, which may possibly mediate increased risk of ocular and systemic vascular complications in these patients. OCTA could potentially be useful as a biomarker for migraine with aura.

Purpose: To evaluate the impact of image magnification correction on superficial retinal vessel density (SRVD) and foveal avascular zone area (FAZA) measurements using optical coherence tomography angiography (OCTA)., Methods: Participants with healthy retinas were recruited for ocular biometry, refraction, and RTVue XR Avanti OCTA imaging with the 3 × 3-mm protocol. The foveal and parafoveal SRVD and FAZA were quantified with custom software before and after correction for magnification error using the Littman and the modified Bennett formulae. Relative changes between corrected and uncorrected SRVD and FAZA were calculated., Results: Forty subjects were enrolled and the median (range) age of the participants was 30 (18-74) years. The mean (range) spherical equivalent refractive error was -1.65 (-8.00 to +4.88) diopters and mean (range) axial length was 24.42 mm (21.27-28.85). Images from 13 eyes were excluded due to poor image quality leaving 67 for analysis. Relative changes in foveal and parafoveal SRVD and FAZA after correction ranged from -20% to +10%, -3% to +2%, and -20% to +51%, respectively. Image size correction in measurements of foveal SRVD and FAZA was greater than 5% in 51% and 74% of eyes, respectively. In contrast, 100% of eyes had less than 5% correction in measurements of parafoveal SRVD., Conclusions: Ocular biometry should be performed with OCTA to correct image magnification error induced by axial length variation. We advise caution when interpreting interocular and interindividual comparisons of SRVD and FAZA derived from OCTA without image size correction.

Purpose: To investigate whether the area and shape of the foveal avascular zone (FAZ) as assessed by optical coherence tomography angiography (OCTA) are altered in glaucomatous eyes with central visual field defects (CVFDs)., Methods: A total of 78 patients with open-angle glaucoma with central or peripheral visual field defects (PVFDs) confined to a single hemifield were studied retrospectively. Foveal avascular zone area and circularity were measured using OCTA images from the superficial retinal layer. Central retinal visual field (VF) sensitivity using Swedish Interactive Threshold Algorithm 24-2 VF and macular ganglion cell-inner plexiform layer (mGCIPL) thickness were measured. The FAZ area between VF-affected hemimacular segments and VF-unaffected hemimacular segments in eyes with CVFDs and matched hemimacular segments of eyes with PVFDs were compared. Factors associated with the presence and severity of CVFD at initial presentation were determined., Results: Eyes with CVFDs showed a significantly larger FAZ area, lower FAZ circularity, and lower mGCIPL thickness than the PVFD group. The mean hemi-FAZ area of VF-affected hemimaculas in eyes with CVFDs was significantly larger than that of the PVFD group (0.256 ± 0.07 mm2 vs. 0.184 ± 0.07 mm2) and the VF-unaffected hemimaculas of the CVFD group (0.179 ± 0.06 mm2; P < 0.05). Age, mean deviation, mGCIPL thickness, FAZ area, and circularity were associated with CVFDs (P < 0.05)., Conclusions: Microcirculatory alterations in the perifovea are spatially correlated with central VF loss. Loss of FAZ circularity was significantly associated with presence of CVFD, whereas FAZ area was significantly associated with severity of CVFD.

Purpose: The purpose of this study was to quantify retinal capillary density and the foveal avascular zone (FAZ) area in normal subjects according to age, using optical coherence tomography angiography (OCTA)., Methods: All eyes in this cross-sectional study underwent OCTA using RTVue XR Avanti with AngioVue. OCTA scans were analyzed and processed, and vessel density and FAZ dimensions were calculated., Results: A total of 113 normal eyes from 70 subjects were included (30 males, 40 females; mean 48 ± 20 years of age). The mean vessel density and FAZ dimensions were significantly smaller in the superficial retinal capillary plexus (SCP) than in the deep retinal capillary plexus (DCP), using quantitative OCTA analysis (all P< 0.0001). With 3 × 3-mm scans, the mean vessel density was 13.431 ± 1.758 mm-1 in the SCP, 18.812 ± 1.796 mm-1 in the DCP, and 5.913 ± 1.308 mm-1 and 10.447 ± 1.262 mm-1 with 6 × 6-mm scans in the SCP and DCP, respectively. Mean FAZ areas were 0.289 ± 0.108 mm2 at the SCP and 0.614 ± 0.200 mm2 at the DCP. Age was a predictor of SCP and DCP vessel density and FAZ area in the SCP. Vessel density decreased 0.0393 mm-1 (0.26%) per year in the SCP and 0.0574 mm-1 (0.27%) per year in the DCP. FAZ areas increased 0.0014 mm2 (0.63%) and 0.0011 mm2 (0.20%) per year in the SCP and DCP, respectively., Conclusions: SCP and DCP vessel density decreased with increasing age, while FAZ area increased with age. Normal age-matched measurements provide important standardized values that may facilitate management of retinal vascular disorders.

Purpose: The primate central retina is characterized by an avascular fovea and well-defined perifoveal capillary plexus. Neither blood vessels nor their accompanying astrocytes enter the fovea during any stage of retinal development; a balance of angiogenic and angiostatic factors probably maintains foveal avascularity throughout life. The aim of this study was to identify potentially angiorepulsive factors involved in the development of the avascular primate retinal fovea., Methods: Retinas of newborn, juvenile, and adult Callithrix jacchus and Macaca fascicularis monkeys and control human retinas were studied to determine the localization of angiostatin relative to III β-tubulin, glial fibrillary acidic protein, vascular endothelial growth factor (VEGF), platelet endothelial cell adhesion molecule-1 (PECAM), and the angiostatin receptor αvβ3-integrin in the foveal, macular, and peripheral retina. Expression studies were performed using immunohistochemistry (IHC) on retinal whole-mount and paraffin sections, and Western blotting on frozen material. The complex network of the main retinal cell types was identified by IHC of retinal whole mounts., Results: In general, lifetime expression of angiostatin was found in all retinas. Colabeling with different markers revealed retinal ganglion cells as the main source of angiostatin expression in the primate retina, whereas PECAM-immunopositive blood capillaries expressed the angiostatin receptor αvβ3-integrin, and capillary-associated astrocytes expressed VEGF., Conclusions: This study provides the first evidence of angiostatin expression in the primate retina; the expression of angiostatin in the avascular foveal region and the peripheral retina suggests that angiostatin may play a role in the regulation of retinal vascularization, providing a possible explanation for the development and persistence of an avascular fovea.

Purpose: To determine the size and characteristics of the superficial and deep foveal avascular zone (FAZ) in healthy adults by using optical coherence tomography angiography (OCT-A), and to ascertain the effects of demographic and ocular parameters on the FAZ size., Methods: In a prospective cohort study of 117 healthy volunteers, foveal-centered 3 × 3-mm OCT-A scans were manually graded by certified graders to determine the size of the superficial and deep FAZ. Multiple linear regression analyses were performed to evaluate the impact of demographics and ocular factors, including central retinal thickness (CRT), choroidal thickness, axial length (AL), and spherical equivalent (SE) on superficial and deep FAZ areas., Results: The mean age of the participants was 22.5 years, with mean AL of 25.4 mm and mean SE of -4.3 diopters. The mean CRT was 262.8 μm (range, 220-316 μm). The mean superficial FAZ area was 0.24 mm2, while the deep FAZ area was 0.38 mm2 (P < 0.001). Females had a larger superficial (P < 0.001) and deep FAZ (P < 0.001). On univariate linear regression, both superficial and deep FAZ areas had significant correlations with CRT, sex, AL, and SE, but not with age. By multiple linear regression analysis, in normal eyes, superficial FAZ area varied significantly with CRT and sex. Among eyes with high myopia, both superficial and deep FAZ varied significantly with CRT, sex, and choroidal thickness., Conclusions: The superficial and deep FAZ areas varied significantly among healthy eyes. Factors such as CRT, sex, SE, AL, and choroidal thickness influence the size of the FAZ.

Purpose: To compare the use of optical coherence tomography angiography (OCTA) and adaptive optics scanning light ophthalmoscope fluorescein angiography (AOSLO FA) for characterizing the foveal microvasculature in healthy and vasculopathic eyes., Methods: Four healthy controls and 11 vasculopathic patients (4 diabetic retinopathy, 4 retinal vein occlusion, and 3 sickle cell retinopathy) were imaged with OCTA and AOSLO FA. Foveal perfusion maps were semiautomatically skeletonized for quantitative analysis, which included foveal avascular zone (FAZ) metrics (area, perimeter, acircularity index) and vessel density in three concentric annular regions of interest. On each set of OCTA and AOSLO FA images, matching vessel segments were used for lumen diameter measurement. Qualitative image comparisons were performed by visual identification of microaneurysms, vessel loops, leakage, and vessel segments., Results: Adaptive optics scanning light ophthalmoscope FA and OCTA showed no statistically significant differences in FAZ perimeter, acircularity index, and vessel densities. Foveal avascular zone area, however, showed a small but statistically significant difference of 1.8% (P = 0.004). Lumen diameter was significantly larger on OCTA (mean difference 5.7 μm, P < 0.001). Microaneurysms, fine structure of vessel loops, leakage, and some vessel segments were visible on AOSLO FA but not OCTA, while blood vessels obscured by leakage were visible only on OCTA., Conclusions: Optical coherence tomography angiography is comparable to AOSLO FA at imaging the foveal microvasculature except for differences in FAZ area, lumen diameter, and some qualitative features. These results, together with its ease of use, short acquisition time, and avoidance of potentially phototoxic blue light, support OCTA as a tool for monitoring ocular pathology and detecting early disease.

Purpose: To demonstrate the utility of speckle variance optical coherence tomography (svOCT), a noninvasive angiographic technique, for evaluating the foveal vasculature., Methods: Twelve normal human eyes were imaged with svOCT (1060-nm, 100-kHz custom-built system) and fluorescein angiography (FA; Topcon TRC-50DX with 5.0 megapixel resolution camera). Manual tracing techniques were used to quantify the foveal vasculature, including foveal avascular zone (FAZ) metrics (area, perimeter, greatest diameter, and lowest diameter). Reproducibility of these measurements was determined. The FAZ was imaged in 25 normal eyes using svOCT and 15 donor eyes using confocal scanning laser microscopy. Retinal capillary plexuses in donor eyes were perfusion-labeled with phalloidin conjugated to Alexa Fluor 546., Results: Speckle variance OCT is able to stratify the foveal circulation into inner and deep capillary plexuses as well as reliably quantify and assess the morphometric dimensions of the human FAZ. Capillary density measurements were significantly greater in svOCT than FA (31.2 ± 1.6% vs. 19.3 ± 1.9% of total tissue area; P < 0.001). Measurements were highly reproducible (all P > 0.366). All FAZ metrics were significantly lower in histology than svOCT (all P < 0.001)., Conclusions: Speckle variance OCT permits precise, reproducible, and noninvasive visualization of the human foveal vasculature. Speckle variance OCT may become an important adjunct in evaluating patients with retinal vascular diseases.

Purpose: We characterized the foveal avascular zone (FAZ) and the parafoveal capillary network in healthy subjects using swept source OCT optical microangiography (OMAG)., Methods: We acquired OMAG images of the macula of 19 eyes (13 healthy individuals) using a prototype swept source laser OCT. En face images of the retinal vasculature were generated for superficial and deep inner retinal layers (SRL/DRL) in regions of interest 250 (ROI-250) and 500 (ROI-500) μm from the FAZ border., Results: The mean area (mm2) of the FAZ was 0.304 ± 0.132 for the SRL and 0.486 ± 0.162 for the DRL (P < 0.001). Mean vessel density (%) was 67.3 ± 6.4 for the SRL and 34.5 ± 8.6 for the DRL in the ROI-250 (P < 0.001), and 74.2 ± 3.9 for the SRL and 72.3 ± 4.9 for the DRL in the ROI-500 (P = 0.160)., Conclusions: Swept source OMAG images of healthy subjects allowed analysis of the FAZ and the density of the parafoveal capillary network at different retinal layers.

Purpose: To investigate macular perfusion in healthy Chinese individuals and examine its dependence on age and sex., Methods: Healthy adult Chinese individuals were recruited. Macular perfusion was measured by spectral-domain optical coherence tomography (OCT) using the split-spectrum amplitude-decorrelation angiography (SSADA) algorithm. The parafoveal flow index and vessel area density as well as the area of the foveal capillary-free zone (CFZ) were quantified., Results: A total of 76 eyes in 45 subjects were included (20 males and 25 females, mean age 36 ± 11 years). The mean parafoveal flow index was 0.099 ± 0.013; the mean vessel area density was 0.891 ± 0.073; and the mean CFZ area was 0.474 ± 0.172 mm2. All three parameters were significantly correlated with age (flow index: P = 0.00; vessel area density: P = 0.00; CFZ area: P = 0.02). The flow index and vessel area density decreased annually by 0.6% and 0.4%, respectively, and CFZ area increased by 1.48% annually. The CFZ area was larger in females than in males, while all three parameters seemed to change more rapidly with age in males than in females., Conclusions: In healthy Chinese eyes, macular perfusion decreased with increasing age, and decreased more rapidly in males than in females. The application of OCT angiograms may provide a useful approach for monitoring macular perfusion, although caution must be exercised with regard to age- and sex-related variations.

Purpose: To determine if patients with cardiovascular comorbidities but no clinically detectable retinal disease demonstrate quantitative alterations to perifoveal capillary networks., Methods: Comparisons were made between 10 eyes from patients with vascular comorbidities and 17 control eyes. All eyes were absent of clinically evident ocular disease. Microcannulation techniques were used to label the retinal microvasculature. Retinae were flat mounted, and the peripapillary region 2 mm nasal to the fovea was imaged using confocal laser scanning microscopy. Two- and three-dimensional image reconstructions were used to perform quantitative measurements of individual capillary networks within the perifovea. Parameters measured included capillary diameter, capillary loop area, capillary loop length, capillary density, and capillary surface area., Results: Capillary diameter was increased in the retinal ganglion cell and superficial inner plexiform layer capillary network in patients with vascular comorbidities. Capillary loop area and capillary loop length were reduced in all capillary networks in patients with vascular comorbidities except the deep capillary network of the inner nuclear layer. Capillary density was reduced in the nerve fiber layer capillary network in patients with vascular comorbidities. There was no difference in the relative occupied capillary surface area between control and diseased eyes., Conclusions: The results in this study suggest that the quantitative characteristics of perifoveal capillary networks are nonuniformly altered in patients with vascular comorbidities, before the onset of clinically identifiable eye diseases. These findings may be important for understanding pathophysiological mechanisms involved in retinal vascular diseases.

Purpose: To investigate erythrocyte aggregates in parafoveal capillaries by adaptive optics scanning laser ophthalmoscopy (AO-SLO)., Methods: AO-SLO videos were acquired from the parafoveal areas of one eye in 10 healthy subjects. Erythrocyte aggregates were detected as "dark tails" that were darker regions than vessel shadows. The lengths of the dark tails were measured in target capillaries, and their time-dependent changes in length were analyzed using spatiotemporal images. The dark tail elongation rate was calculated as the change of dark tail length per unit length of the target capillary., Results: The overall average dark tail length was 112.1 ± 36.9 μm. The dark tail became longer in a time-dependent manner in every monitored capillary (P < 0.0001). The dark tail elongation rate and average velocity were 0.51 ± 0.37 and 1.49 ± 0.36 mm/s, respectively., Conclusions: AO-SLO can be used for noninvasive and direct monitoring of blood dynamics in the retinal microvasculature without dying agents. Erythrocyte aggregates were detected as dark tails and were elongated in a time-dependent manner in the parafoveal capillaries of normal subjects. Monitoring the characteristics of dark tails has promising potential for evaluating retinal hemodynamics.

Purpose: To investigate the relationship between enlargement of the foveal avascular zone (FAZ) and structural changes on spectral-domain optical coherence tomography (SD-OCT) in patients with ischemic diabetic macular edema (DME)., Methods: This was a retrospective, cross-sectional study including 35 eyes of 33 patients with ischemic DME as determined by irregular margins of the FAZ in fluorescein angiography. We measured the best-corrected visual acuity (BCVA), central subfield thickness (CST), subfield volume, subfoveal choroidal thickness, length of the photoreceptor outer segment (PR-OS), and the lateral extent of inner segment-outer segment (IS-OS) disruption on SD-OCT images, as well as the FAZ to optic nerve head (FAZ/ONH) area ratio by using the ImageJ program., Results: The mean FAZ/ONH area ratio was 0.28 ± 0.15. The FAZ/ONH area ratio was positively correlated with the logarithm of the minimum angle of resolution (LogMAR) BCVA (P = 0.001, r = 0.529). The mean CST was 391 ± 110 μm, and the subfield volume was 0.31 ± 0.09 mm(3). The mean length of the PR-OS and the horizontal and vertical extent of IS-OS disruption were 40 ± 8 μm, 169 ± 294 μm, and 114 ± 170 μm, respectively. The mean length of the PR-OS and the horizontal and vertical extent of IS-OS disruption were significantly correlated with FAZ/ONH area ratio (P = 0.020, P = 0.001, and P = 0.049, respectively). The horizontal and vertical extent of IS-OS disruption showed a positive correlation with LogMAR BCVA (P = 0.027 and P = 0.049, respectively)., Conclusions: Foveal ischemia in DME appears to cause PR-OS shortening and IS-OS disruption resulting in outer retinal layer atrophic changes and subsequent visual loss.

Purpose: To quantify the distribution and morphometric characteristics of capillary networks in the human perifovea. To determine correlations between the location of neuronal subcellular compartments and the morphometric features of regional capillary networks in the layered retina., Methods: The perifoveal region, located 2 mm nasal to the fovea, was studied in 17 human donor eyes. Novel micropipette technology was used to cannulate the central retinal artery and label the retinal microcirculation using a phalloidin perfusate. γ-synuclein, Goα, and parvalbumin antibodies were also used to co-localize the nerve fiber layer (NFL), retinal ganglion cell layer (RGCL), inner plexiform layer (IPL), and inner nuclear layer (INL). Confocal scanning laser microscopy was used for capillary imaging. Capillary diameter, capillary density, and capillary loop area measurements were compared between networks., Results: Four capillary networks were identified in the following retinal layers: (1) NFL, (2) RGCL and superficial portion of IPL, (3) deep portion of IPL and superficial portion of INL, and (4) deep portion of INL. Laminar configurations were present in NFL and deep INL networks. Remaining networks demonstrated three-dimensional configurations. Capillary density was greatest in the networks serving the IPL. Capillary loop area was smallest in the two innermost networks. There was no difference in capillary diameter between networks., Conclusions: Capillary networks in the human perifovea are morphometrically heterogeneous. Morphometric features of regional capillary networks in the layered retina may serve a critical role in supporting neuronal homeostasis. Improved knowledge of these features may be important for understanding pathogenic mechanisms underlying retinal vascular diseases.

Purpose: To assess the relationship between foveal pit morphology and size of the foveal avascular zone (FAZ)., Methods: Forty-two subjects were recruited. Volumetric images of the macula were obtained using spectral domain optical coherence tomography. Images of the FAZ were obtained using either a modified fundus camera or an adaptive optics scanning light ophthalmoscope. Foveal pit metrics (depth, diameter, slope, volume, and area) were automatically extracted from retinal thickness data, whereas the FAZ was manually segmented by two observers to extract estimates of FAZ diameter and area., Results: Consistent with previous reports, the authors observed significant variation in foveal pit morphology. The average foveal pit volume was 0.081 mm(3) (range, 0.022 to 0.190 mm(3)). The size of the FAZ was also highly variable between persons, with FAZ area ranging from 0.05 to 1.05 mm(2) and FAZ diameter ranging from 0.20 to 1.08 mm. FAZ area was significantly correlated with foveal pit area, depth, and volume; deeper and broader foveal pits were associated with larger FAZs., Conclusions: Although these results are consistent with predictions from existing models of foveal development, more work is needed to confirm the developmental link between the size of the FAZ and the degree of foveal pit excavation. In addition, more work is needed to understand the relationship between these and other anatomic features of the human foveal region, including peak cone density, rod-free zone diameter, and Henle fiber layer.

Purpose: To report confocal red-free imaging as a novel, noninvasive imaging modality for the detection of the foveal avascular zone (FAZ) and to compare its effectiveness to that of fluorescein angiography (FA) in diabetic retinopathy (DR) and retinal vein occlusion (RVO)., Methods: This was a retrospective, observational, cross-sectional study. The authors enrolled 50 eyes with DR and 20 eyes with RVO. All subjects underwent confocal red-free imaging and FA using a scanning laser ophthalmoscope. For all subjects, the FAZ was identified on both confocal red-free and corresponding FA images and was delineated by two independent readers. The authors evaluated the concordance between the two imaging methods by comparing the sizes of the delineated areas and determining the overlapping ratio after image processing., Results: The image analysis showed a high correlation (r > 0.9) in the mean size of the FAZ between the red-free and corresponding FA images with DR or RVO. Reliable agreement between the two methods was confirmed by the comparison of size (P = 0.925 on independent t-test) and overlapping correspondence (overlapping ratio, 0.77) of the delineated area., Conclusions: The findings suggest that confocal red-free imaging is a simple, reliable, safe, and noninvasive method for effectively imaging the FAZ. This procedure has the potential to be used for the noninvasive detection and quantification of FAZ in screening, initial evaluation, and follow-up observation of progressive ischemic retinopathies such as DR and RVO.

Purpose: To investigate the source of moving particles in parafoveal capillaries detected by adaptive optics scanning laser ophthalmoscopy (AO-SLO)., Methods: AO-SLO videos were acquired from the parafoveal areas of eyes of healthy subjects. The gray-scale values inside and outside the moving particles were measured and compared. Thereafter, successive frames of the captured videos were analyzed under higher magnification to detect changes in the gray values of bright spots inside the capillaries, before and during passage of the particles. Simultaneously, changes in the gray values of areas without the bright spots were measured for comparison. Then, the authors analyzed the packing arrangements of the bright spots in the particles, and measured the particle velocity using spatiotemporal images of the target capillary., Results: There were no significant differences in the gray values between the moving particles and the cone mosaic outside the parafoveal capillaries adjacent to the particles. The gray value of the bright spots in the dark shadow of the vessels increased when the particles passed through, while the dark areas without bright spots remained dark. There were no significant differences in the packing arrangements between the bright spots and surrounding cone mosaic. Further, the concordance rate of packing arrangements of bright spots between two consecutive moving particles in the capillary was 95.8%. The mean particle velocity was 1.34 ± 0.42 mm/s., Conclusions: The particles moving in the capillaries are suggested to be reflections of photoreceptor aggregates that pass through circulating transparent objects such as leukocytes or plasma gaps.

Purpose: To demonstrate the application of phase-variance optical coherence tomography (pvOCT) for contrast agent-free in vivo imaging of volumetric retinal microcirculation in the human foveal region and for extraction of foveal avascular zone dimensions., Methods: A custom-built, high-speed Fourier-domain OCT retinal imaging system was used to image retinas of two healthy subjects and eight diabetic patients. Through the acquisition of multiple B-scans for each scan location, phase differences between consecutive scans were extracted and used for phase-variance contrast, identifying motion signals from within blood vessels and capillaries. The en face projection view of the inner retinal layers segmented out from volumetric pvOCT data sets allowed visualization of a perfusion network with the foveal avascular zone (FAZ). In addition, the authors presented 2D retinal perfusion maps with pseudo color-coded depth positions of capillaries., Results: Retinal vascular imaging with pvOCT provides accurate measurements of the FAZ area and its morphology and a volumetric perfusion map of microcapillaries. In this study using two images from each fundus fluorescein angiography (FA) and pvOCT, the measured average areas of the FAZ from two healthy subjects were below 0.22 mm(2), and each of eight diabetic patients had an enlarged FAZ area, larger than 0.22 mm(2). Moreover, the FAZ areas demonstrated a significant correlation (r = 0.91) between measurements from FA and pvOCT., Conclusions: The high-speed pvOCT allows contrast agent-free visualization of capillary networks in the human foveal region that is analogous to fundus FA imaging. This could allow for noninvasive diagnosis and progression monitoring of diabetic retinopathy in clinical settings.

Purpose: To establish, using adaptive optics scanning laser ophthalmoscopy (AOSLO), that the retinal parafoveal capillary network is altered before the onset of diabetic retinopathy in adult patients with type 2 diabetes., Methods: AOSLO videos were acquired in the parafoveal region of one eye from control subjects and from patients with type 2 diabetes and no retinopathy. Detailed images of the parafoveal capillary network were generated with custom motion contrast enhancement algorithms. The combination of AOSLO images and videos enabled the simultaneous assessment of several features of the parafoveal capillary network. Arteriovenous (AV) channels were identified by finding the least tortuous capillary channels connecting terminal arterioles to postcapillary venules. Measures of capillary dropout and capillary hemodynamics were also quantified., Results: The average tortuosity of AV channels was 26% higher in patients with type 2 diabetes when compared with controls, even though there were no signs of diabetic retinopathy in any of the eyes that were assessed (P < 0.05). In addition, the metrics of capillary dropout showed small changes (between 3% and 7%), leukocyte speed 14% lower, and pulsatility 25% higher, but none of these differences was statistically significant., Conclusions: It is often difficult to find consistent changes in the retinal microvasculature due to large intersubject variability. However, with a novel application of AOSLO imaging, it is possible to visualize parafoveal capillaries and identify AV channels noninvasively. AV channels are disrupted in type 2 diabetes, even before the onset of diabetic retinopathy.

Purpose: Ultrahigh-resolution optical coherence tomography (UHR-OCT) with adaptive optics (AO) provides micrometer-scale 3D resolution that is attractive for imaging the retinal microvasculature. Such imaging may be useful for early detection of pathologic changes as in diabetic retinopathy. Here the authors investigate this potential for detecting individual capillaries in healthy subjects., Methods: UHR-AO-OCT volumes centered on the fovea were acquired from seven subjects (age range, 25-61 years) with three preselected with no foveal avascular zone (FAZ). Images were compared with entoptic diagrams using the capillaries at the rim of the FAZ. Methods of comparison were testing for the presence of a FAZ, noting distinct features in the capillary pattern, and measuring the size of the FAZ. Additional analysis included measurements of capillary diameter and depth range with retinal eccentricity., Results: UHR-AO-OCT results are consistent with entoptic observations for all three methods of comparison. FAZ diameters measured by UHR-AO-OCT and entoptic imaging are strongly correlated (R(2) = 0.86). Average capillary diameter near the FAZ rim is 5.1 (4.6) ± 1.4 μm, with the value in parentheses accounting for axial image blur. This is consistent with histology (average, ~4.7 μm). Depth range of the capillaries increases monotonically with eccentricity (0°-1.25°) and is larger and more variable for subjects without FAZ., Conclusions: UHR-AO-OCT permits observation of many of the capillaries proximal to the FAZ, including those of average size based on published histology. This supports the view that the vast majority of capillaries in the retina are likely detectable with UHR-AO-OCT.

Purpose: The shape of the human fovea presents important but still poorly characterized variations. In this study, the variability of the shape and structure of normal foveae were examined., Methods: In a group of 110 eyes of 57 healthy adults, the shape and structure of the fovea were analyzed by automated segmentation of retinal layer on high-resolution optical coherence tomography scans. In an additional group of 10 normal eyes of 10 patients undergoing fluorescein angiography, the size of the foveal avascular zone (FAZ) was correlated to foveal shape., Results: From the thickest to the thinnest fovea, there was a structural continuum ranging from a shallow pit with continuity of the inner nuclear layer (INL) over the center (seven eyes; 6.7%), to a complete separation of inner layers overlying a flat and thinner central outer nuclear layer (ONL; eight eyes; 7.3%). Central foveal thickness correlated inversely to the degree of inner layer separation and to the surface of the FAZ., Conclusions: Foveal structure strongly correlates with its neurovascular organization. The findings support a developmental model in which the size of the FAZ determines the extent of centrifugal migration of inner retinal layers, which counteracts in some way the centripetal packing of cone photoreceptors.

Purpose: To demonstrate noninvasive imaging of human foveal capillary networks with a high-resolution, wide-field, dual-conjugate adaptive optics (DCAO) imaging instrument., Methods: The foveal capillary networks of five healthy subjects with no previous history of ocular or neurologic disease or surgery were imaged with a novel high-resolution, wide-field DCAO instrument. The foveal avascular zone (FAZ) in each image was defined using a manual procedure. An automated algorithm based on publicly available and custom-written software was used to identify vessels and extract morphologic FAZ and vessel parameters. Capillary densities were calculated in two annular regions of interest (ROIs) outside the FAZ (500 μm and 750 μm outer radius from the foveal center) and in the superior, inferior, temporal, and nasal quadrants within the two ROIs., Results: Mean FAZ area was 0.302 ± 0.100 mm(2), and mean capillary density (length/area) in the inner ROI was 38.0 ± 4.0 mm(-1) and 36.4 ± 4.0 mm(-1) in the outer ROI. The difference in ROI capillary density was not significant. There was no significant difference in quadrant capillary density within the two ROIs or between quadrants irrespective of ROI., Conclusions: The authors have demonstrated a technique for noninvasive imaging and semiautomated detection and analysis of foveal capillaries. In comparison with other studies, their method yielded lower capillary densities than histology but similar results to the current clinical gold standard, fluorescein angiography. The increased field of view of the DCAO instrument opens up new possibilities for high-resolution noninvasive clinical imaging of foveal capillaries.

Purpose: To assess the effects of intravitreal bevacizumab injections in the treatment of subfoveal choroidal neovascularization (CNV) associated with pattern dystrophy (PD) of the retinal pigment epithelium., Methods: The study was a prospective, nonrandomized, open-label, interventional clinical trial in which 12 patients were prospectively enrolled. Patients with a diagnosis of PD complicated by subfoveal CNV were considered for the study. All patients underwent a complete ophthalmic examination, including ETDRS visual acuity measurement, electroretinogram, electrooculogram, optical coherence tomography, and fluorescein angiography. The treatment protocol began with a loading dose of three consecutive injections at 1-month intervals, followed by injections administered as needed, according to OCT parameters and angiographic features observed during a 24-month follow-up period. The number of eyes with a visual acuity loss of fewer than 15 letters (<3 ETDRS lines), compared with baseline measures, was recorded at the 6-, 12-, and 24-month examinations., Results: Twelve patients completed the planned visits and were included in the study. A visual acuity loss of fewer than 15 letters was not registered in any case at the 6- and 12-month examinations and was found in only one (8%) patient at the 24-month examination. The mean best corrected visual acuity (BCVA) and the mean central macular thickness (CMT) at baseline were 0.73+/-0.34 (logMAR+/-SD) and 276+/-95 microm (SD), respectively. At the 3-month examination, the mean BCVA significantly improved to 0.48+/-0.27, whereas the mean CMT decreased to 220+/-71 microm. At the 12-month examination, the mean BCVA was 0.45+/-0.24, and the mean CMT was 209+/-53 microm. At the 24-month (last) follow-up, the BCVA showed substantial stabilization and the CMT decreased to 199+/-34 microm. No side effects or complications were registered., Conclusions: Intravitreal bevacizumab injection is a beneficial treatment for subfoveal CNV associated with PD. Further studies are warranted to confirm these initial results and to analyze the morphofunctional changes during the follow-up. (ClinicalTrials.gov number, NCT00391144.).

PURPOSE. To describe and evaluate the performance of a computerized automated segmentation technique for use in quantification of the foveal avascular zone (FAZ). METHODS. A computerized technique for automated segmentation of the FAZ using images from fundus fluorescein angiography (FFA) was applied to 26 transit-phase images obtained from patients with various grades of diabetic retinopathy. The area containing the FAZ zone was first extracted from the original image and smoothed by a Gaussian kernel (sigma = 1.5). An initializing contour was manually placed inside the FAZ of the smoothed image and iteratively moved by the segmentation program toward the FAZ boundary. Five tests with different initializing curves were run on each of 26 images to assess reproducibility. The accuracy of the program was also validated by comparing results obtained by the program with the FAZ boundaries manually delineated by medical retina specialists. Interobserver performance was then evaluated by comparing delineations from two of the experts. RESULTS. One-way analysis of variance indicated that the disparities between different tests were not statistically significant, signifying excellent reproducibility for the computer program. There was a statistically significant linear correlation between the results obtained by automation and manual delineations by experts. CONCLUSIONS. This automated segmentation program can produce highly reproducible results that are comparable to those made by clinical experts. It has the potential to assist in the detection and management of foveal ischemia and to be integrated into automated grading systems.

Purpose: To demonstrate a noninvasive method to visualize and analyze the parafoveal capillary network in humans., Methods: An adaptive optics scanning laser ophthalmoscope was used to acquire high resolution retinal videos on human subjects. Video processing tools that enhance motion contrast were developed and applied to the videos to generate montages of parafoveal retinal capillaries. The capillary network and foveal avascular zone (FAZ) were extracted using video and image analysis algorithms. The capillary densities in the zone immediately outside the FAZ were calculated and the variation in density as a function of direction was investigated. Extracted FAZ geometries were used to calculate area and effective diameters. The authors also compared their method against fluorescein angiography (FA) for one subject., Results: The parafoveal capillaries were clearly visible when the motion contrast in noninvasive videos was enhanced. There was a marked improvement in the contrast of the parafoveal capillaries when compared to the unprocessed videos. The average FAZ area was 0.323 mm(2), with an average effective diameter of 633 microm. There was no variation in capillary density near the FAZ in different directions., Conclusions: Using motion cues to enhance vessel contrast is a powerful tool for visualizing the capillary network, in the absence of contrast agents. The authors demonstrate a tool to study the microcirculation of healthy subjects noninvasively.