The Curvature, Collagen Network Structure, and Their Relationship to the Pressure-Induced Strain Response of the Human Lamina Cribrosa in Normal and Glaucoma Eyes.

The lamina cribrosa (LC) is a connective tissue in the optic nerve head (ONH). The objective of this study was to measure the curvature and collagen microstructure of the human LC, compare the effects of glaucoma and glaucoma optic nerve damage, and investigate the relationship between the structure and pressure-induced strain response of the LC in glaucoma eyes. Previously, the posterior scleral cups of 10 normal eyes and 16 diagnosed glaucoma eyes were subjected to inflation testing with second harmonic generation (SHG) imaging of the LC and digital volume correlation (DVC) to calculate the strain field. In this study, we applied a custom microstructural analysis algorithm to the maximum intensity projection of SHG images to measure features of the LC beam and pore network. We also estimated the LC curvatures from the anterior surface of the DVC-correlated LC volume. Results showed that the LC in glaucoma eyes had larger curvatures p≤0.03), a smaller average pore area (p = 0.001), greater beam tortuosity (p < 0.0001), and more isotropic beam structure (p = 0.01) than in normal eyes. The difference measured between glaucoma and normal eyes may indicate remodeling of the LC with glaucoma or baseline differences that contribute to the development of glaucomatous axonal damage.

Mechanical strain in the mouse astrocytic lamina increases after exposure to recombinant trypsin.

The responses of astrocytes in the optic nerve head (ONH) to mechanical and biochemical stimuli are important to understanding the degeneration of retinal ganglion cell axons in glaucoma. The ONH in glaucoma is vulnerable to stress produced by the intraocular pressure (IOP). Notably, after three days of elevated IOP in a mouse model, the junctions between the astrocytic processes and the peripapillary sclera were altered and the structural compliance of the ONH increased. In order to simulate this aspect of glaucomatous remodeling, explanted mouse eyes were treated with TrypLE, a recombinant trypsin enzyme. Treatment with TrypLE caused the periphery of the astrocytic lamina to contract radially by 0.044 ± 0.038. Transmission electron microscopy showed that TrypLE caused a separation of the end-feet of the astrocyte processes from the basement membrane at the junction with the sclera. Inflation testing after treatment with TrypLE caused an increased strain response in the astrocytic lamina compared to the strain response before treatment. The greatest increase was in the radial Green-Lagrange strain, Err = 0.028 ± 0.009, which increased by 340%. The alterations in the microstructure and in the strain response of the astrocytic lamina reported in mouse experimental glaucoma were partially reproduced by experimental treatment of mouse eyes with TrypLE. The results herein suggest that separation of junctions between the astrocyte processes and the sclera may be instrumental in increasing the structural compliance of the ONH after a period of elevated IOP. STATEMENT OF SIGNIFICANCE: Astrocytes of the optic nerve of the eye spread out from edge to edge across the optic nerve in a region referred to as the astrocytic lamina. In an experimental model of glaucoma caused by elevated eye-pressure, there is disruption of the connections between astrocytes and the edge of the astrocytic lamina. We caused a similar event in the lamina by incubating explanted mouse eyes with an enzyme. Disruption of the astrocyte connections to the edge of their tissue caused the tissue to stretch more when we increased the eye-pressure, compared to the control tissue. This work is the first on the tissue of the optic nerve to demonstrate the importance of cell connections in preventing the over-stretching of the astrocytic lamina.

Ion-Complex Microcrystal Formulation Provides Sustained Delivery of a Multimodal Kinase Inhibitor from the Subconjunctival Space for Protection of Retinal Ganglion Cells.

Glaucoma is the leading cause of irreversible blindness worldwide. Elevated intraocular pressure (IOP) is one of the major risk factors for glaucoma onset and progression, and available pharmaceutical interventions are exclusively targeted at IOP lowering. However, degeneration of retinal ganglion cells (RGCs) may continue to progress despite extensive lowering of IOP. A complementary strategy to IOP reduction is the use of neuroprotective agents that interrupt the process of cell death by mechanisms independent of IOP. Here, we describe an ion complexation approach for formulating microcrystals containing ~50% loading of a protein kinase inhibitor, sunitinib, to enhance survival of RGCs with subconjunctival injection. A single subconjunctival injection of sunitinib-pamoate complex (SPC) microcrystals provided 20 weeks of sustained retina drug levels, leading to neuroprotection in a rat model of optic nerve injury. Furthermore, subconjunctival injection of SPC microcrystals also led to therapeutic effects in a rat model of corneal neovascularization. Importantly, therapeutically relevant retina drug concentrations were achieved with subconjunctival injection of SPC microcrystals in pigs. For a chronic disease such as glaucoma, a formulation that provides sustained therapeutic effects to complement IOP lowering therapies could provide improved disease management and promote patient quality of life.

Biomechanics of the optic nerve head and peripapillary sclera in a mouse model of glaucoma.

The deformation of the mouse astrocytic lamina (AL) and adjacent peripapillary sclera (PPS) was measured in response to elevated intraocular pressure. We subjected explanted mouse eyes to inflation testing, comparing control eyes to those 3 days and 6 weeks after induction of ocular hypertension (OHT) via ocular microbead injection. Laser scanning microscopy was used with second harmonic generation to image the collagenous PPS and two-photon fluorescence to image transgenic fluorescent astrocytes in the AL. Digital volume correlation was applied to calculate strains in the PPS and AL. The specimen-averaged strains were biaxial in the AL and PPS, with greater strain overall in the x- than y-direction in the AL and greater strain in the θ- than the r-direction in the PPS. Strains increased after 3-day OHT, with greater strain overall in the 3-day AL than control AL, and greater circumferential strain in the 3-day PPS than control PPS. In the 6-week OHT eyes, AL and PPS strains were similar overall to controls. This experimental glaucoma model demonstrated a dynamic change in the mechanical behaviour of the AL and PPS over time at the site of neuronal injury and remodelling in glaucoma.

Pressure-Induced Changes in Astrocyte GFAP, Actin, and Nuclear Morphology in Mouse Optic Nerve.

Purpose: To conduct quantitative analysis of astrocytic glial fibrillary acidic protein (GFAP), actin and nuclei distribution in mouse optic nerve (ON) and investigate changes in the measured features after 3 days of ocular hypertension (OHT). Method: Serial cross-sections of 3-day microbead-induced OHT and control ONs were fluorescently labelled and imaged using confocal microscope. Eighteen structural features were measured from the acquired images, including GFAP coverage, actin area fraction, process thickness, and aspect ratio of cell nucleus. The measured features were analyzed for variations with axial locations along ON and radial zones transverse to ON, as well as for the correlations with degree of intraocular pressure (IOP) change. Results: The most significant changes in structural features after 3-day OHT occurred in the unmyelinated ON region (R1), and the changes were greater with greater IOP elevation. Although the GFAP, actin, axonal, and ON areas all increased in 3-day OHT ONs in R1 (P ≤ 0.004 for all), the area fraction of GFAP actually decreased (P = 0.02), the actin area fraction was stable and individual axon compartments were unchanged in size. Within R1, the number of nuclear clusters increased (P < 0.001), but the mean size of nuclear clusters was smaller (P = 0.02) and the clusters became rounder (P < 0.001). In all cross-sections of control ONs, astrocytic processes were thickest in the rim zone compared with the central and peripheral zones (P ≤ 0.002 for both), whereas the overall process width in R1 decreased after 3 days of OHT (P < 0.001). Conclusions: The changes in structure elucidated IOP-generated alterations that underlie astrocyte mechanotranslational responses relevant to glaucoma.

The Effects of Glaucoma on the Pressure-Induced Strain Response of the Human Lamina Cribrosa.

Purpose: To measure the ex vivo pressure-induced strain response of the human optic nerve head and analyze for variations with glaucoma diagnosis and optic nerve axon damage. Methods: The posterior sclera of 16 eyes from 8 diagnosed glaucoma donors and 10 eyes from 6 donors with no history of glaucoma were inflation tested between 5 and 45 mm Hg. The optic nerve from each donor was examined for degree of axon loss. The posterior volume of the lamina cribrosa (LC) was imaged with second harmonic generation and analyzed using volume correlation to calculate LC strains between 5 and 10 and 5 and 45 mm Hg. Results: Eye length and LC area were larger in eyes diagnosed with glaucoma (P ≤ 0.03). Nasal-temporal EXX and circumferential Eθθ strains were lower in the LC of diagnosed glaucoma eyes at 10 mm Hg (P ≤ 0.05) and 45 mm Hg (P ≤ 0.07). EXX was smaller in the LC of glaucoma eyes with <25% axon loss compared with undamaged normal eyes (P = 0.01, 45 mm Hg). In general, the strains were larger in the peripheral than central LC. The ratio of the maximum principal strain Emax in the peripheral to central LC was larger in glaucoma eyes with >25% axon loss than in glaucoma eyes with milder damage (P = 0.004, 10 mm Hg). Conclusions: The stiffness of the LC pressure-strain response was greater in diagnosed glaucoma eyes and varied with glaucomatous axon damage. Lower LC strains in glaucoma eyes with milder damage may represent baseline biomechanical behavior that contributes to axon loss, whereas greater LC strain and altered radial LC strain variation in glaucoma eyes with more severe damage may be caused by glaucoma-related remodeling.

The inflation response of the human lamina cribrosa and sclera: Analysis of deformation and interaction.

This study investigated the inflation response of the lamina cribrosa (LC) and adjacent peripapillary sclera (PPS) in post-mortem human eyes with no history of glaucoma. The posterior sclera of 13 human eyes from 7 donors was subjected to controlled pressurization between 5-45 mmHg. A laser-scanning microscope (LSM) was used to image the second harmonic generation (SHG) response of collagen and the two-photon fluorescent (TPF) response of elastin within the volume of the LC and PPS at each pressure. Image volumes were analyzed using digital volume correlation (DVC) to calculate the three-dimensional (3D) deformation field between pressures. The LC exhibited larger radial strain, Err, and maximum principal strain, Emax, (p < 0.0001) and greater posterior displacement (p=0.0007) compared to the PPS between 5-45 mmHg, but had similar average circumferential strain, Eθθ, and maximum shear strain, Γmax. The Emax and Γmax were highest near the LC-PPS interface and lowest in the nasal quadrant of both tissues. Larger LC area was associated with smaller Emax in the peripheral LC and larger Emax in the central LC (p ≤ 0.01). The Emax, Γmax, and Eθθ in the inner PPS increased with increasing strain in adjacent LC regions (p ≤ 0.001). Smaller strains in the PPS were associated with a larger difference in the posterior displacement between the PPS and central LC (p < 0.0001 for Emax and Err), indicating that a stiffer pressure-strain response of the PPS is associated with greater posterior bowing of the LC. STATEMENT OF SIGNIFICANCE: Glaucoma causes vision loss through progressive damage of the retinal ganglion axons at the lamina cribrosa (LC), a connective tissue structure that supports the axons as they pass through the eye wall. It is hypothesized that strains caused by intraocular pressure may initiate this damage and that these strains are modulated by the combined deformation of the LC and adjacent peripapillary sclera (PPS). In this study we present a method to measure the pressure-induced 3D displacement and strain field in the LC and PPS simultaneously. Regional strain variation in the LC and PPS was investigated and compared and strains were analyzed for associations with age, LC area, LC strain magnitude, and LC posterior motion relative to the PPS.

Characterizing the Collagen Network Structure and Pressure-Induced Strains of the Human Lamina Cribrosa.

Purpose: The purpose of this study was to measure the 2D collagen network structure of the human lamina cribrosa (LC), analyze for the correlations with age, region, and LC size, as well as the correlations with pressure-induced strains. Methods: The posterior scleral cups of 10 enucleated human eyes with no known ocular disease were subjected to ex vivo inflation testing from 5 to 45 mm Hg. The optic nerve head was imaged by using second harmonic generation imaging (SHG) to identify the LC collagen structure at both pressures. Displacements and strains were calculated by using digital volume correlation of the SHG volumes. Nine structural features were measured by using a custom Matlab image analysis program, including the pore area fraction, node density, and beam connectivity, tortuosity, and anisotropy. Results: All strain measures increased significantly with higher pore area fraction, and all but the radial-circumferential shear strain (Erθ) decreased with higher node density. The maximum principal strain (Emax) and maximum shear strain (Γmax) also increased with larger beam aspect ratio and tortuosity, respectively, and decreased with higher connectivity. The peripheral regions had lower node density and connectivity, and higher pore area fraction, tortuosity, and strains (except for Erθ) than the central regions. The peripheral nasal region had the lowest Emax, Γmax, radial strain, and pore area fraction. Conclusions: Features of LC beam network microstructure that are indicative of greater collagen density and connectivity are associated with lower pressure-induced LC strain, potentially contributing to resistance to glaucomatous damage.

Detection of Age-Related Macular Degeneration by Portable Optical Coherence Tomography Operated by Nonexpert Personnel: Potential Use for Screenings.

PURPOSE: The purpose of this study is to determine the sensitivity and specificity of detecting age-related macular degeneration (AMD) using portable optical coherence tomography (OCT) operated by nonexpert photographers on undilated patients. METHODS: In this case-control study, 92 individuals were recruited from the glaucoma and retina clinics at the Wilmer Eye Institute (Johns Hopkins University, Baltimore, MD). Using the portable iVue (Optovue, Inc, Fremont, CA) spectral-domain OCT (SD-OCT), 2 nonexpert photographers acquired retina map scans on undilated eyes of all participants. In total, 33 AMD eyes and 105 control eyes were evaluated and graded by ophthalmologists masked to the diagnoses. RESULTS: Detection of AMD on the portable OCT by ophthalmologists exhibited sensitivities of 0.91 and 0.94 and specificities of 0.88 and 0.89, for graders 1 and 2, respectively. A strong intergrader agreement was observed (κ = 0.87). CONCLUSIONS: Nonexpert photographers can use a portable OCT on undilated eyes to acquire images for the detection of AMD. These findings present the potential utility of implementing a portable OCT in community screenings for earlier detection and treatment of disease.

Evaluation of Automated Segmentation Algorithms for Optic Nerve Head Structures in Optical Coherence Tomography Images.

Purpose: To compare the identification of optic nerve head (ONH) structures in optical coherence tomography images by observers and automated algorithms. Methods: ONH images in 24 radial scan sets by optical coherence tomography were obtained in 51 eyes of 29 glaucoma patients and suspects. Masked intraobserver and interobserver comparisons were made of marked endpoints of Bruch's membrane opening (BMO) and the anterior lamina cribrosa (LC). BMO and LC positional markings were compared between observer and automated algorithm. Repeated analysis on 20 eyes by the algorithm was compared. Regional ONH data were derived from the algorithms. Results: Intraobserver difference in BMO width was not significantly different from zero (P ≥ 0.32) and the difference in LC position was less than 1% different (P = 0.04). Interobserver were slightly larger than intraobserver differences, but interobserver BMO width difference was 0.36% (P = 0.63). Mean interobserver difference in LC position was 14.74 µm (P = 0.004), 3% of the typical anterior lamina depth (ALD). Between observer and algorithm, BMO width differed by 1.85% (P = 0.23) and mean LC position was not significantly different (3.77 µm, P = 0.77). Repeat algorithmic analysis had a mean difference in BMO area of 0.38% (P = 0.47) and mean ALD difference of 0.54 ± 0.72%. Regional ALD had greater variability in the horizontal ONH regions. Some individual outlier images were not validly marked by either observers or algorithm. Conclusions: Automated identification of ONH structures is comparable to observer markings for BMO and anterior LC position, making BMO a practical reference plane for algorithmic analysis.

The Contribution of Sulfated Glycosaminoglycans to the Inflation Response of the Human Optic Nerve Head.

Purpose: In this study, we measured the effect of the removal of sulfated glycosaminoglycans (sGAGs) on the pressure-induced strains of the human lamina cribrosa (LC). Methods: We applied an ex vivo inflation method to measure the three-dimensional (3D) deformation response of six human LCs to pressure, before and after the degradation of chondroitin and dermatan sulfates. The experiment used a laser-scanning microscope (LSM) to acquire the second harmonic generation (SHG) signal of the collagen structure in the LC. Digital volume correlation (DVC) was used to calculate the deformation in the LC after a change in pressure from 5 to 45 mm Hg. Results: The average strains between 5 and 45 mm Hg in the LC decreased significantly after sGAG degradation (P ≤ 0.03), with the greatest change occurring in regions of previously high strain (P ≤ 0.003) and the peripheral regions of the LC (P ≤ 0.02). The stiffening effect was greater in the LC of middle-aged (42-49 years) donors compared with those of older (64-88 years) donors (P < 0.0001). Conclusions: The LC experienced less strain at the same pressures after most sGAGs were removed. These results suggest that the natural decrease in sGAGs within the LC with age may contribute to the stiffer inflation response of older LC to IOP. Likewise, the increase in the amount of sGAGs observed in the LC of glaucomatous eyes, may contribute to a more compliant LC, which may affect the susceptibility and progression of axon damage.

The effects of age on mitochondria, axonal transport, and axonal degeneration after chronic IOP elevation using a murine ocular explant model.

The purpose of this study was to compare younger and older mice after chronic intraocular pressure (IOP) elevation lasting up to 4 days with respect to mitochondrial density, structure, and movement, as well as axonal integrity, in an ex vivo explant model. We studied 2 transgenic mouse strains, both on a C57BL/6J background, one expressing yellow fluorescent protein (YFP) in selected axons and one expressing cyan fluorescent protein (CFP) in all mitochondria. Mice of 4 months or 14 months of age were exposed to chronic IOP by anterior chamber microbead injection for 14 h, 1, 3, or 4 days. The optic nerve head of globe--optic nerve explants were examined by laser scanning microscopy. Mitochondrial density, structure, and movement were quantified in the CFP explants, and axonal integrity was quantified in YFP explants. In control mice, there was a trend towards decreased mitochondrial density (# per mm2) with age when comparing younger to older, control mice, but this was not significant (1947 ±â€¯653 vs 1412 ±â€¯356; p = 0.19). Mitochondrial density decreased after IOP elevation, significantly, by 31%, in younger mice (p = 0.04) but trending towards a decrease, by 22%, in older mice (p = 0.82) compared to age matched controls. Mitochondrial mean size was not altered after chronic IOP elevation for 14 h or more (p ≥ 0.16). When assessing mitochondrial movement, in younger mice, 5% were mobile at any given time; 4% in the anterograde direction and 1% retrograde. In younger untreated tissue, only 75% of explants had moving mitochondria (mean = 15.8 moving/explant), while after glaucoma induction only 24% of explants had moving mitochondria (mean = 4.2 moving/explant; difference from control, p = 0.03). The distance mitochondria traveled in younger mice was unchanged after glaucoma exposure, but in older glaucoma explants the distance traveled was less than half of older controls (p < 0.0003). In younger mice, mitochondrial speed increased after 14 h of elevated IOP (p = 0.006); however, in older glaucoma explants, movement was actually slower than controls (p = 0.02). In RGC-YFP explants, axonal integrity declined significantly after 4 days of IOP elevation to a similar degree in both younger and older mice. Older mice underwent greater loss of mitochondrial movement with chronic IOP elevation than younger mice, but suffered similar short-term axonal fragmentation in C57BL/6J mice. These transgenic strains, studied in explants, permit observations of alterations in intracellular structure and organelle activity in experimental glaucoma.

Use of Optical Coherence Tomography by Nonexpert Personnel as a Screening Approach for Glaucoma.

PURPOSE: This pilot study was conducted to assess optical coherence tomography (OCT) as a screening tool for glaucoma when used by nonexpert personnel. METHODS: This prospective case-control study included 54 patients with open-angle glaucoma and 54 age-matched comparison individuals. Optovue iVue SD-OCT imaging was performed by nonprofessional photographers on undilated patients. The sensitivity, specificity, negative predictive value, and positive predictive value of iVue scan parameters for detecting open-angle glaucoma were evaluated. RESULTS: The iVue cup to disc vertical ratio had a sensitivity of 0.96 [95% confidence interval (CI), 0.90-1.00] at 90% specificity and was strongly correlated with both the Cirrus HD-OCT cup to disc vertical ratio (Pearson coefficient=0.84) and the cup to disc ratio observed on dilated clinical examination by faculty ophthalmologists (Pearson coefficient=0.80). The retinal nerve fiber layer (RNFL) parameters performed robustly, but the ganglion cell complex parameters showed limited diagnostic value. The inferior quadrant thickness was among the best performing RNFL parameters, with a sensitivity of 0.87 (95% CI, 0.78-0.96) and a specificity of 0.88 (95% CI, 0.80-0.97) using the iVue normative database thresholds for abnormality. CONCLUSIONS: OCT imaging may be performed by nonprofessional photographers on undilated patients, and quantitative parameters derived from the resultant images, particularly the vertical cup to disc ratio and the RNFL inferior quadrant thickness, demonstrate sensitivities and specificities that may be adequately robust for glaucoma screening in the community setting.

A Protective Eye Shield Reduces Limbal Strain and Its Variability During Simulated Sleep in Adults With Glaucoma.

PURPOSE: To determine the effect of wearing a protective eye shield (mask) on limbal strain magnitude and variability in glaucoma eyes when sleeping with 1 side of the face down (FD) against a pillow. METHODS: A prospective, randomized, interventional trial was conducted at the Wilmer Eye Institute with 36 glaucoma patients. A contact lens sensor measured limbal strain (output in equivalent millivolts) during intervals of up to 60 minutes in lateral decubitus, FD, and supine positions. Eighteen subjects wore a mask during 1 of 2 FD intervals, with randomized assignment of the interval. Data from additional trials with no mask were included in some analyses. In addition, some facial-feature dimensions from 3D scanned images of 23 subjects were compared with limbal strain data. RESULTS: Wearing a mask trends toward a reduced mean change in contact lens sensor output (limbal strain) on moving to a FD positions [+34.1 mVeq, P=0.01 reduced by -22.3 mVeq, P=0.09 (n=36)]. Mask wearing reduced variability in strain while FD [-22.8 mVeq, P=0.04 (n=18)]. In eyes with past progressive visual field loss, the effect of the mask reduced mean strain change when moving to FD [-44.8 mVeq, P=0.02 (n=31)]. Longer corneal apex to nose-tip and to temple lengths were associated with reduced variability while FD [P=0.02 and 0.04, respectively (n=23)]. Treating both lengths as confounding factors increased statistical significance, particularly for analysis of the no-mask change in strain data moving to and from the FD position [P=0.004 to 0.002 and P=0.03 to 0.01 (n=23)]. CONCLUSION AND RELEVANCE: Wearing a mask reduced limbal strain and variation in limbal strain during simulated FD sleep, particularly in eyes with past field worsening, as did some facial features.

Regional Retinal Ganglion Cell Axon Loss in a Murine Glaucoma Model.

Purpose: To determine if retinal ganglion cell (RGC) axon loss in experimental mouse glaucoma is uniform in the optic nerve. Methods: Experimental glaucoma was induced for 6 weeks with a microbead injection model in CD1 (n = 78) and C57BL/6 (B6, n = 68) mice. From epoxy-embedded sections of optic nerve 1 to 2 mm posterior to the globe, total nerve area and regional axon density (axons/1600 µm2) were measured in superior, inferior, nasal, and temporal zones. Results: Control eyes of CD1 mice have higher axon density and more total RGCs than control B6 mice eyes. There were no significant differences in control regional axon density in all mice or by strain (all P > 0.2, mixed model). Exposure to elevated IOP caused loss of RGC in both strains. In CD1 mice, axon density declined without significant loss of nerve area, while B6 mice had less density loss, but greater decrease in nerve area. Axon density loss in glaucoma eyes was not significantly greater in any region in either mouse strain (both P > 0.2, mixed model). In moderately damaged CD1 glaucoma eyes, and CD1 eyes with the greatest IOP elevation exposure, density loss differed by region (P = 0.05, P = 0.03, mixed model) with the greatest loss in the temporal and superior regions, while in severely injured B6 nerves superior loss was greater than inferior loss (P = 0.01, mixed model, Bonferroni corrected). Conclusions: There was selectively greater loss of superior and temporal optic nerve axons of RGCs in mouse glaucoma at certain stages of damage. Differences in nerve area change suggest non-RGC responses differ between mouse strains.

Visual function quality of life measure changes upon conversion to neovascular age-related macular degeneration in second eyes.

PURPOSE: To determine changes in quality of life measures when choroidal neovascularization (CNV) developed in the second eye of patients with initially unilateral neovascular age-related macular degeneration (AMD). METHODS: We analyzed responses to the 39-item National Eye Institute Visual Function Questionnaire (NEI-VFQ), 36-item Short Form Health Survey (SF-36), and Hospital Anxiety and Depression Scale (HADS) at baseline, and prior to and following second eye CNV diagnosis in 92 participants enrolled in two Submacular Surgery Trials. Paired t-tests for sample sizes over 30 and Wilcoxon signed-rank tests for sample sizes <30 were performed to compare scores. RESULTS: CNV development resulted in statistically and clinically significant changes in responses to 20 of 39 NEI-VFQ items, indicating visual function decline during a mean interval of 25 months. Little difference was noted between baseline scores and prior to CNV diagnosis, which averaged 8.9 months duration. Subscales demonstrated a statistically significant decline in general vision, near activities, distance activities, social functioning, role difficulties, dependency, and driving. There were minimal changes in the HADS and SF-36 scales. CONCLUSION: CNV development in the second eye had a dramatic effect on visual functioning based on patient responses to the NEI-VFQ questionnaire. Our investigation is believed to be the first study using data collected prospectively to demonstrate vision-related quality of life changes that resulted from development of CNV in AMD patients.

Measuring Deformation in the Mouse Optic Nerve Head and Peripapillary Sclera.

Purpose: To develop an ex vivo explant system using multiphoton microscopy and digital volume correlation to measure the full-field deformation response to intraocular pressure (IOP) change in the peripapillary sclera (PPS) and in the optic nerve head (ONH) astrocytic structure. Methods: Green fluorescent protein (GFP)-glutamate transporter-GLT1 (GLT1/GFP) mouse eyes were explanted and imaged with a laser-scanning microscope under controlled inflation. Images were analyzed for regional strains and changes in astrocytic lamina and PPS shape. Astrocyte volume fraction in seven control GLT1/GFP mice was measured. The level of fluorescence of GFP fluorescent astrocytes was compared with glial fibrillary acidic protein (GFAP) labeled astrocytes using immunohistochemistry. Results: The ONH astrocytic structure remained stable during 3 hours in explants. Control strain-globally, in the central one-half or two-thirds of the astrocytic lamina-was significantly greater in the nasal-temporal direction than in the inferior-superior or anterior-posterior directions (each P≤ 0.03, mixed models). The PPS opening (perimeter) in normal eye explants also became wider nasal-temporally than superior-inferiorly during inflation from 10 to 30 mm Hg (P = 0.0005). After 1 to 3 days of chronic IOP elevation, PPS area was larger than in control eyes (P = 0.035), perimeter elongation was 37% less than controls, and global nasal-temporal strain was significantly less than controls (P = 0.007). Astrocyte orientation was altered by chronic IOP elevation, with processes redirected toward the longitudinal axis of the optic nerve. Conclusions: The explant inflation test measures the strain response of the mouse ONH to applied IOP. Initial studies indicate regional differences in response to both acute and chronic IOP elevation within the ONH region.

The pressure-induced deformation response of the human lamina cribrosa: Analysis of regional variations.

The objective of this study was to measure the pressure-induced deformation response of the human lamina cribrosa (LC) and analyze for variations with age and anatomical region. The posterior scleral cup of 8 eyes from 6 human donors was mounted onto a custom inflation chamber. A laser-scanning microscope was used for second harmonic generation (SHG) imaging of the collagen structure in the posterior volume of the LC at pressures from 5mmHg to 45mmHg. The SHG volumes were analyzed by the Fast-Fourier Iterative Digital Volume Correlation (DVC) algorithm for the three dimensional (3D) displacement field. The components of the Green-Lagrange strain tensor and the in-plane principal and maximum shear strains were evaluated from the DVC displacement field for the central and peripheral regions of the LC and the nasal, temporal, inferior, and superior quadrants surrounding the central retinal artery and vein. Among the major findings were that older age was associated with lower strains, the maximum shear strain was larger in the peripheral than central region, and the maximum principal strain was lower in the nasal quadrant. The elliptical shape of the LC was also predictive of the biaxial strain ratio. Age-related and structure-related variations in the pressure-induced strains of the LC may contribute to the susceptibility and severity of optic nerve damage in glaucoma, and regional variations may explain the progression of axonal damage and tissue remodeling observed in the LC in glaucoma. STATEMENT OF SIGNIFICANCE: Glaucoma causes vision loss through progressive damage of the retinal ganglion axons at the lamina cribrosa (LC), the connective tissue structure that supports the axons as they leave the eye. Mechanical characterization of the LC is challenging because of the complex 3D shape and inaccessibility of the tissue. We present a new method using digital volume correlation to map the 3D displacement and strain fields in the LC under inflation. We report for the first time significant regional variations in the strains that are consistent with the pattern of optic nerve damage in early glaucoma. Thus regional strain variations may be predictive of the progression of axonal damage in glaucoma.

Measured Changes in Limbal Strain During Simulated Sleep in Face Down Position Using an Instrumented Contact Lens in Healthy Adults and Adults With Glaucoma.

IMPORTANCE: Eyes of patients with glaucoma may be damaged during sleep. OBJECTIVE: To measure strains in glaucoma eyes and control eyes produced by mechanical force or deformation of the eye from contact when one side of the face rests against a pillow. DESIGN, SETTING, AND PARTICIPANTS: This study took place in a clinic-based setting among 22 patients with glaucoma and 11 age-matched controls. The research was conducted at Wilmer Eye Institute between February 4, 2014, and December 2, 2014. Data analysis was done from June 3, 2014, to June 30, 2015. MAIN OUTCOMES AND MEASURES: We used a contact lens sensor (CLS) to measure change in limbal strain associated with placing one side of the face down (FD) on a pillow in simulated sleep. Baseline intraocular pressure (IOP) was measured with a tonometer. The CLS data were collected every 5 minutes during intervals of up to 60 minutes in various positions, including sitting, lateral decubitus, FD (with the CLS-instrumented eye toward the pillow), and supine. Measured changes in limbal strain were related to estimated changes in IOP and to modeled strain produced by changes in IOP. RESULTS: Among 22 patients with glaucoma and 11 controls, 17 were female. The mean age for the glaucoma group was 62.6 years, while the mean age for the control group was 61.4 years (P = .68). Baseline IOP was also similar for the 2 groups. The mean IOP sitting at the start was 13.7 mm Hg for the glaucoma group and 13.8 mm Hg for the control group (P = .73), and the mean IOP lying at the start was 17.5 mm Hg for the glaucoma group and 16.0 mm Hg for the control group (P = .88). By multivariable linear regression, FD position was associated with an increase in limbal strain in glaucoma eyes (mean [SE], 44.1 [20.4] mV Eq; P = .03) but not in control eyes (mean [SE], 13.6 [13.9] mV Eq, P = .33). While FD, the increased CLS values in patients with glaucoma did not decrease over time (slope, 0.275 mV Eq/min; P = .53 by univariable linear regression). Magnitudes of measured changes in limbal strain were greater in glaucoma eyes with past visual field worsening (P = .006 by multivariable linear modeling). The mean limbal strain increase among patients with glaucoma in FD position was equivalent to strain expected for a mean (SE) IOP increase of 2.5 (1.1) mm Hg from a baseline IOP of 14.2 mm Hg. CONCLUSIONS AND RELEVANCE: Contact with a pillow in FD position during simulated sleep produced a sustained strain increase in glaucoma eyes, particularly those eyes with past progressive visual field loss. The mean FD change in glaucoma eyes was equivalent to strain increase associated with a mean (SE) sustained IOP elevation of 2.5 (1.1) mm Hg. Further experiments are planned to determine if facial features or a protective eye shield prevents sleep position-induced limbal strains during a mean 8-hour sleep period.

Adherence With the Use of Target Intraocular Pressure for Glaucoma Patients in a Large University Practice.

PURPOSE: To determine how often glaucoma specialists set a target intraocular pressure (IOP) and how they respond when the target is not achieved. METHODS: We reviewed 250 randomly selected charts of glaucoma patients seen by 5 glaucoma specialists to identify whether a target IOP was specifically set and to detail the plan of action when the target was exceeded. RESULTS: A target IOP was present for at least 1 eye in 66% of patients (165/250). Among the patients of 5 physicians, the percentage with a target IOP for both eyes ranged from 90% to 34%. Half of eyes with no target had an explaining feature, whereas the other half did not. The study visit IOP exceeded the target in at least 1 eye in 29% (50/173) of patients. When the target was not met, 66% (33/50) had a notation of action taken, with no significant difference among physicians in whether any action was taken (P=0.64). The significant differences among the 5 physicians in the use of target IOP were potentially associated with patient demographic and clinical features, such as age, race, treatment intensity, and visual field damage, which differed among the 5 practices (all P<0.05). CONCLUSIONS: Target IOP was recorded in the large majority of patient charts, but its use varied by physician and patient characteristics. On nearly one third of visits, IOP exceeded the target, indicating the need for active decision-making and management changes.