Healthy Human Eyes Research Articles

Robust and accurate corneal interfaces segmentation in 2D and 3D OCT images.

Segmentation of corneal layer interfaces in optical coherence tomography (OCT) images is important for diagnostic and surgical purposes, while manual segmentation is a time-consuming and tedious process. This paper presents a novel technique for the automatic segmentation of corneal layer interfaces using customized initial layer estimation and a gradient-based segmentation method. The proposed method was also extended to three-dimensional OCT images. Validation was performed on two corneal datasets, one with 37 B-scan images of healthy human eyes and the other with a 3D volume scan of a porcine eye. The approach showed robustness in extracting different layer boundaries in the low-SNR region with lower computational cost but higher accuracy compared to existing techniques. It achieved segmentation errors below 2.1 pixels for both the anterior and posterior layer boundaries in terms of mean unsigned surface positioning error for the first dataset and 2.6 pixels (5.2 μm) for segmenting all three layers that can be resolved in the second dataset. On average, it takes 0.7 and 0.4 seconds to process a cross-sectional B-scan image for datasets one and two, respectively. Our comparative study also showed that it outperforms state-of-the-art methods for quantifying layer interfaces in terms of accuracy and time efficiency.

Extended and adjustable field-of-view of variable interscan time analysis by ammonite-scanning swept-source optical coherence tomography angiography.

A novel scanning protocol, ammonite scan, is proposed for widefield optical coherence tomography angiography (OCTA) and relative retinal blood flow velocity imaging in the human retina using variable interscan time analysis (VISTA). A repeated circle scan using a 400 kHz swept-source was employed to achieve an interscan time of 1.28 ms. The center of the repeated circular scan continuously moved spirally towards the peripheral region, ensuring an extended and adjustable scan range while preserving the short interscan time. Image artifacts due to eye movement were eliminated via extra motion-correction processing using data redundancy. The relative blood flow velocity in superficial and deep plexus layers was calculated from the VISTA image, and their ratio was used to explore the microvascular flow parameter in the healthy human eye.

Exploiting multi-granularity visual features for retinal layer segmentation in human eyes.

Accurate segmentation of retinal layer boundaries can facilitate the detection of patients with early ophthalmic disease. Typical segmentation algorithms operate at low resolutions without fully exploiting multi-granularity visual features. Moreover, several related studies do not release their datasets that are key for the research on deep learning-based solutions. We propose a novel end-to-end retinal layer segmentation network based on ConvNeXt, which can retain more feature map details by using a new depth-efficient attention module and multi-scale structures. In addition, we provide a semantic segmentation dataset containing 206 retinal images of healthy human eyes (named NR206 dataset), which is easy to use as it does not require any additional transcoding processing. We experimentally show that our segmentation approach outperforms state-of-the-art approaches on this new dataset, achieving, on average, a Dice score of 91.3% and mIoU of 84.4%. Moreover, our approach achieves state-of-the-art performance on a glaucoma dataset and a diabetic macular edema (DME) dataset, showing that our model is also suitable for other applications. We will make our source code and the NR206 dataset publicly available at (https://github.com/Medical-Image-Analysis/Retinal-layer-segmentation).

Whole-genome sequencing of Paracoccus species isolated from the healthy human eye and description of Paracoccus shanxieyensis sp. nov.

Currently, the genus Paracoccus comprises 76 recognized species. Members of Paracoccus are mostly isolated from environmental, animal, and plant sources. This report describes and proposes a novel species of Paracoccus isolated from clinical specimens of the human ocular surface. We isolated two aerobic, Gram-stain-negative, non-spore-forming, coccoid or short rod-shaped, and non-motile strains (designated DK398T and DK608) from conjunctival sac swabs of two healthy volunteers. The results showed that the strains grew best under the conditions of 28°C, pH 7.0, and 1.0 % (w/v) NaCl. Sequence analysis based on the 16S rRNA gene showed that strains DK398T and DK608 were members of Paracoccus, most similar to Paracoccus laeviglucosivorans 43PT (98.54 and 98.62 %), Paracoccus litorisediminis GHD-05T (98.34 and 98.41 %), and Paracoccus limmosus NB88T (98.21 and 98.29 %). Phenotypic analysis showed that DK398T and DK608 were positive for catalase and oxidase, negative for producing N-acetyl-β-glucosaminic acid, arginine dihydrolase, and β-glucuronidase but positive for leucine arylamidase. The predominant isoprenoid quinone was Q-10, and the major polar lipids included phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, and an unidentified glycolipid. The major fatty acids (>10%) were summed feature 8 (C18 : 1 ω7c and/or C18 : 1 ω6c) and C16 : 0. The meso-diaminopimelic acid was found in the cell wall peptidoglycan of DK398T. The major cell wall sugars were ribose and galactose. Based on the results of phylogenetic analyses, low (<83.22 %) average nucleotide identity, digital DNA-DNA hybridization (<26.0%), chemotaxonomic analysis, and physiological properties, strain DK398T represents a novel species of the genus Paracoccus, for which the name Paracoccus shanxieyensis sp. nov. is proposed. The type strain is DK398T (=CGMCC 1.17227T=JCM 33719T).

Typical localised element-specific finite element anterior eye model

PurposeThe study presents an averaged anterior eye geometry model combined with a localised material model that is straightforward, appropriate and amenable for implementation in finite element (FE) modelling. MethodsBoth right and left eye profile data of 118 subjects (63 females and 55 males) aged 22–67 years (38.5 ± 7.6) were used to build an averaged geometry model. Parametric representation of the averaged geometry model was achieved through two polynomials dividing the eye into three smoothly connected volumes. This study utilised the collagen microstructure x-ray data of 6 ex-vivo healthy human eyes, 3 right eyes and 3 left eyes in pairs from 3 donors, 1 male and 2 females aged between 60 and 80 years, to build a localised element-specific material model for the eye. ResultsFitting the cornea and the posterior sclera sections to a 5th-order Zernike polynomial resulted in 21 coefficients. The averaged anterior eye geometry model recorded a limbus tangent angle of 37° at a radius of 6.6 mm from the corneal apex. In terms of material models, the difference between the stresses generated in the inflation simulation up to 15 mmHg in the ring-segmented material model and localised element-specific material model were significantly different (p < 0.001) with the ring-segmented material model recording average Von-Mises stress 0.0168 ± 0.0046 MPa and the localised element-specific material model recording average Von-Mises stress 0.0144 ± 0.0025 MPa. ConclusionsThe study illustrates an averaged geometry model of the anterior human eye that is easy to generate through two parametric equations. This model is combined with a localised material model that can be used either parametrically through a Zernike fitted polynomial or non-parametrically as a function of the azimuth angle and the elevation angle of the eye globe. Both averaged geometry and localised material models were built in a way that makes them easy to implement in FE analysis without additional computation cost compared to the limbal discontinuity so-called idealised eye geometry model or ring-segmented material model.

Exploiting Multi-Granularity Visual Features for Retinal Layer Segmentation in Healthy Human Eyes

Exploiting Multi-Granularity Visual Features for Retinal Layer Segmentation in Healthy Human Eyes

Modeling the Endothelial Glycocalyx Layer in the Human Conventional Aqueous Outflow Pathway.

A layer of proteoglycans and glycoproteins known as glycocalyx covers the surface of the trabecular meshwork (TM), juxtacanalicular tissue (JCT), and Schlemm's canal (SC) inner wall of the conventional aqueous outflow pathway in the eye. This has been shown to play a role in the mechanotransduction of fluid shear stress and in the regulation of the outflow resistance. The outflow resistance in the conventional outflow pathway is the main determinant of the intraocular pressure (IOP) through an active, two-way, fluid-structure interaction coupling between the outflow tissues and aqueous humor. A 3D microstructural finite element (FE) model of a healthy human eye TM/JCT/SC complex with interspersed aqueous humor was constructed. A very thin charged double layer that represents the endothelial glycocalyx layer covered the surface of the elastic outflow tissues. The aqueous humor was modeled as electroosmotic flow that is charged when it is in contact with the outflow tissues. The electrical-fluid-structure interaction (EFSI) method was used to couple the charged double layer (glycocalyx), fluid (aqueous humor), and solid (outflow tissues). When the IOP was elevated to 15 mmHg, the maximum aqueous humor velocity in the EFSI model was decreased by 2.35 mm/s (9%) compared to the fluid-structure interaction (FSI) model. The charge or electricity in the living human conventional outflow pathway generated by the charged endothelial glycocalyx layer plays a minor biomechanical role in the resultant stresses and strains as well as the hydrodynamics of the aqueous humor.

The Effect of Intraocular Pressure Load Boundary on the Biomechanics of the Human Conventional Aqueous Outflow Pathway.

Aqueous humor outflow resistance in the trabecular meshwork (TM), juxtacanalicular connective tissue (JCT), and Schlemm's canal (SC) endothelium of the conventional outflow pathway actively contribute to intraocular pressure (IOP) regulation. Outflow resistance is actively affected by the dynamic outflow pressure gradient across the TM, JCT, and SC inner wall tissues. The resistance effect implies the presence of a fluid-structure interaction (FSI) coupling between the outflow tissues and the aqueous humor. However, the biomechanical interactions between viscoelastic outflow tissues and aqueous humor dynamics are largely unknown. A 3D microstructural finite element (FE) model of a healthy human eye TM/JCT/SC complex was constructed with elastic and viscoelastic material properties for the bulk extracellular matrix and embedded elastic cable elements. The FE models were subjected to both idealized and a physiologic IOP load boundary using the FSI method. The elastic material model for both the idealized and physiologic IOP load boundary at equal IOPs showed similar stresses and strains in the outflow tissues as well as pressure in the aqueous humor. However, outflow tissues with viscoelastic material properties were sensitive to the IOP load rate, resulting in different mechanical and hydrodynamic responses in the tissues and aqueous humor. Transient IOP fluctuations may cause a relatively large IOP difference of ~20 mmHg in a very short time frame of ~0.1 s, resulting in a rate stiffening in the outflow tissues. Rate stiffening reduces strains and causes a rate-dependent pressure gradient across the outflow tissues. Thus, the results suggest it is necessary to use a viscoelastic material model in outflow tissues that includes the important role of IOP load rate.

Viscoelastic Biomechanical Properties of the Conventional Aqueous Outflow Pathway Tissues in Healthy and Glaucoma Human Eyes.

Background: Although the tissues comprising the ocular conventional outflow pathway have shown strong viscoelastic mechanical response to aqueous humor pressure dynamics, the viscoelastic mechanical properties of the trabecular meshwork (TM), juxtacanalicular connective tissue (JCT), and Schlemm’s canal (SC) inner wall are largely unknown. Methods: A quadrant of the anterior segment from two human donor eyes at low- and high-flow (LF and HF) outflow regions was pressurized and imaged using optical coherence tomography (OCT). A finite element (FE) model of the TM, the adjacent JCT, and the SC inner wall was constructed and viscoelastic beam elements were distributed in the extracellular matrix (ECM) of the TM and JCT to represent anisotropic collagen. An inverse FE-optimization algorithm was used to calculate the viscoelastic properties of the ECM/beam elements such that the TM/JCT/SC model and OCT imaging data best matched over time. Results: The ECM of the glaucoma tissues showed significantly larger time-dependent shear moduli compared to the heathy tissues. Significantly larger shear moduli were also observed in the LF regions of both the healthy and glaucoma eyes compared to the HF regions. Conclusions: The outflow tissues in both glaucoma eyes and HF regions are stiffer and less able to respond to dynamic IOP.

Identifying Collector Channel Orifices In Vivo with Phase-Sensitive Optical Coherence Tomography: A Preliminary Study

Collector channels are openings located in the trabecular meshwork (TM) of the human eye that function as conduits, connecting the anterior chamber to the episcleral veins. Identifying the positions of collector channel orifices (CCOs) is essential for positioning implants in microinvasive canal-based glaucoma surgery, which is still not possible in vivo currently. Considerable evidence indicates that aqueous outflow becomes more active near the CCOs. Because the TM movement regulates the aqueous outflow, identification of the TM motion signal has the potential to locate the CCOs. Phase-sensitive optical coherence tomography (PhS-OCT) is an effective tool for the instantaneous detection of TM motion in vivo with sensitivity at the nanometer scale. However, the downside of this method is that phase measurement is prone to mix noises that negatively distinguish between biomedical signals. The TM motion was considered initially to be set up by the cardiac pulse. In this paper, a signal quality index related to blood pressure monitoring was applied to assess the validity of the TM motion signal. Measurements were carried out on two pairs of healthy human eyes. Quantitative measurements of the TM motion signal region such as size and frequency were recorded as the judgment indicator for CCOs. These results demonstrate that the PhS-OCT is a valuable tool capable of revealing the aqueous outflow pathway in vivo, offering a novel alternative to optimize glaucoma surgery.

Correlation Between Laser Speckle Flowgraphy and OCT-Derived Retinal and Choroidal Metrics in Healthy Human Eye.

PurposeTo investigate the correlation between laser speckle flowgraphy (LSFG) signals and the quantitative metrics derived from optical coherence tomography (OCT) in normal eyes.MethodsLSFG, OCT, and OCT angiography (OCTA) imaging were performed on normal participants using a custom-designed LSFG system and a commercial swept-source OCT system. Mean (PWM) and amplitude (PWA) of the LSFG pulse waveform were selected to quantify the LSFG signals. Retinal and choroidal maps were obtained using the standard 6 × 6 mm OCT and OCTA scans. Structural and vascular metrics maps, including thickness, vessel area density, vessel skeleton density, and vessel diameter index of the retina, and choroidal thickness (CT), choroidal vessel volume (CVV) and choroidal vessel index (CVI), were employed to quantify the retinal and choroidal properties. Correlation analysis was then performed between the LSFG, retinal, and choroidal metrics maps.ResultsTwelve healthy participants aged 23 to 36 years were enrolled in this study. The spatial distribution of the PWM and PWA values was highly correlated with that of the CT and CVV metrics. On average, Spearman correlation coefficients (ρ) were 0.80 and 0.78 (all P < 0.001) for the correlations between PWM and CT and CVV, respectively, and were 0.61 and 0.63 (all P < 0.05) for the correlations between PWA and CT and CVV, respectively. In comparison, both PWM and PWA were generally weak or not correlated with all the retinal metrics and CVI.ConclusionsLSFG signals were positively correlated with the choroidal thickness and vessel volume, suggesting choroidal blood flows dominate the LSFG signals at the area absent of large retinal vessels.Translational RelevanceThis study illustrates the dominant source of the LSFG signals in the eye.

The Human Eye Transcriptome Atlas: A searchable comparative transcriptome database for healthy and diseased human eye tissue.

The applications of deep sequencing technologies in life science research and clinical diagnostics have increased rapidly over the last decade. Although fast algorithms for data processing exist, intuitive, portable solutions for data analysis are still rare. For this purpose, we developed a web-based transcriptome database, which provides a platform-independent, intuitive solution to easily explore and compare ocular gene expression of 100 diseased and healthy human tissue samples from 15 different tissue types collected at the Eye Center of the University of Freiburg. To ensure comparability of expression between different tissues, reads were normalized across all 100 samples. Differentially expressed genes were calculated between each tissue type to determine tissue-specific genes. Unsupervised analysis of all 100 samples revealed an accurate clustering according to different tissue types and a high tissue specificity by analyzing known tissue-specific marker genes. Bioinformatic cell type deconvolution using xCell provided detailed insights into the cellular profiles of each tissue type. Several new tissue-specific marker genes were identified. These genes were involved in tissue- or disease-specific processes, such as myelination for the optic nerve, visual perception for retina, keratinocyte differentiation for conjunctival carcinoma, as well as endothelial cell migration for choroidal neovascularization membranes. The results are accessible at the Human Eye Transcriptome Atlas website at https://www.eye-transcriptome.com. In summary, this searchable transcriptome database enables easy exploration of ocular gene expression in healthy and diseased human ocular tissues without bioinformatics expertise. Thus, it provides rapid access to detailed insights into the molecular mechanisms of various ocular tissues and diseases, as well as the rapid retrieval of potential new diagnostic and therapeutic targets.

Visual photosensitivity threshold and objective photosensitivity luminance in healthy human eyes assessed using an automated ocular photosensitivity analyser: a step towards translation of a clinical tool for assessing photophobia.

To evaluate the visual photosensitivity threshold and objective photosensitivity luminance in healthy eyes, thereby providing a normative dataset that will lead to a better understanding of diseases causing photophobia. This was a prospective cross-sectional study. Emmetropes whose visual acuity was better than 0.18logMAR (6/9) with no other ocular abnormality were included. Headache Impact Test-6 and visual light sensitivity questionnaires were administered. Visual photosensitivity threshold was measured subjectively using the ocular photosensitivity analyser. Objective photosensitivity luminance was assessed manually by evaluating videos recorded using an infrared camera and noting the intensity of light at the first squeezing reflex. Seventy five normal subjects (age range, 7-71years) were included. Median age was 32.7years (inter-quartile range, 20.3-47.9years). Forty (53.3%) were males. Median Headache Impact Test score was 38 (inter-quartile range, 36-42) and visual light sensitivity questionnaire score was 11 (inter-quartile range, 8-15). Mean (standard deviation) right eye, left eye and binocular visual photosensitivity threshold was 3.34 (0.78), 3.33 (0.81) and 3.37 (0.78) loglux, respectively. There was a significant negative correlation of visual light sensitivity questionnaire scores with right eye, left eye and binocular visual photosensitivity thresholds, and positive correlation of age with binocular visual photosensitivity thresholds. Mean (standard deviation) right eye, left eye and binocular objective photosensitivity luminance was 3.25 (0.55), 3.35 (0.47) and 3.15 (0.52) loglux, respectively. Age was only positively correlated with binocular objective photosensitivity luminance, and there was no correlation between age and right eye or left eye objective photosensitivity luminance. The study characterised, for the first time, objective photosensitivity luminance and established normative data for both visual photosensitivity threshold and objective photosensitivity luminance. The data will help in understanding the pathophysiology of diseases causing photophobia, monitoring the disease progression and evaluating treatment modalities.

Ocular Surface Temperature: Characterization in a Large Cohort of Healthy Human Eyes and Correlations to Systemic Cardiovascular Risk Factors.

Purpose: To characterize ocular surface temperature (OST) in healthy eyes and its association with systemic risk factors of cardiovascular and ischemic heart disease. Methods: This prospective cross-sectional study included consenting subjects who were examined at the Institute for Medical Screening in Sheba Medical Center. A Therm-App™ thermal imaging camera (Opgal LTD, Israel) was used for OST acquisition, and the mean OST of the medial canthal, lateral canthal, and central cornea regions were measured. Room and body temperatures were also recorded. Past medical and ocular history as well as data from various clinical examinations performed at the same visit were obtained. Results: Thermographic images were obtained from 186 subjects, 150 of which were included in the final analysis. OST was significantly higher in the medial canthal, central cornea, and lateral canthal regions in people with a history of ischemic heart disease (p = 0.02, p = 0.02, and p = 0.03, respectively). There were no significant OST differences (ANOVA test) associated with the presence of hypertension, diabetes mellitus, or active smoking status. Conclusions: OST correlated positively with the presence of ischemic heart disease. This correlation, its pathophysiological base, and its clinical application warrants further investigation.

Comparative study of deep neural networks with unsupervised Noise2Noise strategy for noise reduction of optical coherence tomography images.

As a powerful diagnostic tool, optical coherence tomography (OCT) has been widely used in various clinical setting. However, OCT images are susceptible to inherent speckle noise that may contaminate subtle structure information, due to low-coherence interferometric imaging procedure. Many supervised learning-based models have achieved impressive performance in reducing speckle noise of OCT images trained with a large number of noisy-clean paired OCT images, which are not commonly feasible in clinical practice. In this article, we conducted a comparative study to investigate the denoising performance of OCT images over different deep neural networks through an unsupervised Noise2Noise (N2N) strategy, which only trained with noisy OCT samples. Four representative network architectures including U-shaped model, multi-information stream model, straight-information stream model and GAN-based model were investigated on an OCT image dataset acquired from healthy human eyes. The results demonstrated all four unsupervised N2N models offered denoised OCT images with a performance comparable with that of supervised learning models, illustrating the effectiveness of unsupervised N2N models in denoising OCT images. Furthermore, U-shaped models and GAN-based models using UNet network as generator are two preferred and suitable architectures for reducing speckle noise of OCT images and preserving fine structure information of retinal layers under unsupervised N2N circumstances.

Diurnal Variation of Retinal Vessel Density in Healthy Human Eyes.

A small increase in optic nerve head vessel density (VD), but not macular VD, in the evening compared with the morning is observed in healthy subjects. To evaluate the diurnal variation of the macular and optic nerve head (ONH) VD in healthy eyes as measured with optical coherence tomography angiography (OCT-A). In this prospective study of healthy individuals older than 18 years old, VD parameters, including macular whole image vessel density, parafoveal vessel density, ONH whole image vessel density, ONH whole image capillary density, circumpapillary vessel density (cpVD), and circumpapillary capillary density, were measured with OCT-A at 4 time points throughout the day (8 am, 12 pm, 4 pm, and 8 pm). Twenty-nine healthy eyes were included from 15 subjects (mean age: 30.9 y). After adjustment for age and mean ocular perfusion pressure, a significant positive rate of change was found for cpVD (0.05%/h; P=0.027). In contrast, macular VD changes were not significantly different. When comparing morning (8 am and 12 pm) and evening (4 pm and 8 pm) measurements, there were small, but statistically significant, increases for all ONH measurements. The greatest increase was found for cpVD (0.58%; P=0.022). Significant but small increases in mean arterial pressure and mean ocular perfusion pressure were also observed. There was a small increase in ONH VD, but not macular VD, in the evening compared with the morning in healthy human eyes. As the observed difference was within the measurement variability, our results suggest the timing of OCT-A examination does not affect retinal VD measurements.

Gaze-Dependent Simulation of Light Perception in Virtual Reality.

The perception of light is inherently different inside a virtual reality (VR) or augmented reality (AR) simulation when compared to the real world. Conventional head-worn displays (HWDs) are not able to display the same high dynamic range of brightness and color as the human eye can perceive in the real world. To mimic the perception of real-world scenes in virtual scenes, it is crucial to reproduce the effects of incident light on the human visual system. In order to advance virtual simulations towards perceptual realism, we present an eye-tracked VR/AR simulation comprising effects for gaze-dependent temporal eye adaption, perceptual glare, visual acuity reduction, and scotopic color vision. Our simulation is based on medical expert knowledge and medical studies of the healthy human eye. We conducted the first user study comparing the perception of light in a real-world low-light scene to a VR simulation. Our results show that the proposed combination of simulated visual effects is well received by users and also indicate that an individual adaptation is necessary, because perception of light is highly subjective.

Label-free adaptive optics imaging of human retinal macrophage distribution and dynamics

Microglia are resident central nervous system macrophages and the first responders to neural injury. Until recently, microglia have been studied only in animal models with exogenous or transgenic labeling. While these studies provided a wealth of information on the delicate balance between neuroprotection and neurotoxicity within which these cells operate, extrapolation to human immune function has remained an open question. Here we examine key characteristics of retinal macrophage cells in live human eyes, both healthy and diseased, with the unique capabilities of our adaptive optics-optical coherence tomography approach and owing to their propitious location above the inner limiting membrane (ILM), allowing direct visualization of cells. Our findings indicate that human ILM macrophage cells may be distributed distinctly, age differently, and have different dynamic characteristics than microglia in other animals. For example, we observed a macular pattern that was sparse centrally and peaked peripherally in healthy human eyes. Moreover, human ILM macrophage density decreased with age (∼2% of cells per year). Our results in glaucomatous eyes also indicate that ILM macrophage cells appear to play an early and regionally specific role of nerve fiber layer phagocytosis in areas of active disease. While we investigate ILM macrophage cells distinct from the larger sample of overall retinal microglia, the ability to visualize macrophage cells without fluorescent labeling in the live human eye represents an important advance for both ophthalmology and neuroscience, which may lead to novel disease biomarkers and new avenues of exploration in disease progression.

Acute Effects of Intraocular Pressure-Induced Changes in Schlemm's Canal Morphology on Outflow Facility in Healthy Human Eyes.

PurposeTo estimate the outflow facility coefficient (C) as a function of Schlemm's canal cross-sectional area (SCAR) in healthy subjects using noninvasive oculopression tonometry (OPT).MethodsIn 25 healthy volunteers, intraocular pressure (IOP) decay values were recorded by a ophthalmodynamometer, with a fixed external force (0.15 N) on the inferior-temporal eyelid, every 10 seconds, for four minutes, and again after a 30-minute rest. Schlemm's canal profile images and IOP were obtained pre-procedurally (baseline), immediately (T0), and at 1-minute intervals post-procedurally (T1, T2, T3, and T4). C was calculated for different IOPs. The SCAR, coronal, and the meridional diameter of Schlemm's canal were calculated.ResultsMean C0 for the maximum IOP was 0.020 ± 0.017 µL/min/mm Hg; mean C was 0.018 ± 0.0071 and 0.058 ± 0.0146 µL/min/mm Hg at 40 and 20 mm Hg, respectively. C was nonlinearly dependent on the IOP (R2 = 0.945). The SCAR was 5440 ± 3140.82, 3947.6 ± 2246.8, and 5375.7 ± 2662.7 µm2 at baseline, T0, and T4, respectively. The coronal diameter of SC decreased significantly from the baseline (33.02 ± 11.3 µm) to T0 (26.6 ± 9.37 µm) and recovered at T4 (32.3 ± 9.53 µm). The SCAR and IOP correlated significantly throughout (R2 = 0.9944; P < 0.001). C0 significantly correlated with the SCAR at baseline and with changes in the SCAR and IOP from T0 to T4.ConclusionsSchlemm's canal dimensions are responsible for the IOP-dependent mechanical forces, and these changes appear to directly affect outflow facility.

Retinal Distribution and Extracellular Activity of Granzyme B: A Serine Protease That Degrades Retinal Pigment Epithelial Tight Junctions and Extracellular Matrix Proteins.

Granzymes are a family of serine proteases first shown to be intracellular initiators of immune-mediated cell death in target pathogenic cells. In addition to its intracellular role, Granzyme B (GzmB) has important extracellular functions in immune regulation and extracellular matrix (ECM) degradation. Verified substrates of extracellular GzmB activity include tight junctional and ECM proteins. Interestingly, little is known about the activity of GzmB in the outer human retina, a tissue in which the degradation of the tight junctional contacts of retinal pigment epithelial (RPE) cells and within the external limiting membrane, as well as remodeling of the ECM in Bruch's membrane, cause the breakdown of the blood-retinal barrier and slowing of metabolite transport between neuroretina and choroidal blood supply. Such pathological changes in outer retina signal early events in the development of age-related macular degeneration (AMD), a multifactorial, chronic inflammatory eye disease. This study is the first to focus on the distribution of GzmB in the outer retina of the healthy and diseased post-mortem human eye. Our results revealed that GzmB is present in RPE and choroidal mast cells. More immunoreactive cells are present in older (>65 years) compared to younger (<55 years) donor eyes, and choroidal immunoreactive cells are more numerous in eyes with choroidal neovascularization (CNV), while RPE immunoreactive cells are more numerous in eyes with soft drusen, an early AMD event. In vitro studies demonstrated that RPE-derived tight junctional and ECM proteins are cleaved by exogenous GzmB stimulation. These results suggest that the increased presence of GzmB immunoreactive cells in outer retina of older (healthy) eyes as well as in diseased eyes with CNV (from AMD) and eyes with soft drusen exacerbate ECM remodeling in the Bruch's membrane and degradation of the blood-retinal barrier. Currently there are no treatments that prevent remodeling of the Bruch's membrane and/or the loss of function of the outer blood-retinal barrier, known to promote early AMD changes, such as drusen deposition, RPE dysfunction and pro-inflammation. Specific inhibitors of GzmB, already in preclinical studies for non-ocular diseases, may provide new strategies to stop these early events associated with the development of AMD.