Showing papers by "Harry A. Quigley published in 2018"

Rho-Kinase Inhibition Reduces Myofibroblast Differentiation and Proliferation of Scleral Fibroblasts Induced by Transforming Growth Factor β and Experimental Glaucoma.

Abstract: Purpose We evaluated prevention of transforming growth factor β (TGFβ)-induced transdifferentiation of cultured scleral fibroblasts to myofibroblasts by rho-associated protein kinase (ROCK) inhibitors. Additionally, we tested whether local delivery of ROCK inhibitors reduced scleral fibroblast proliferation in response to chronic intraocular pressure (IOP) elevation. Methods Primary human peripapillary sclera (PPS) fibroblasts were cultured and treated with TGFβ to induce myofibroblast transdifferentiation, as determined by immunoblot assessment of α smooth muscle actin (SMA) levels and collagen gel contraction. Cells were treated with the ROCK inhibitors Y27632, fasudil, and H1152 before TGFβ treatment. ROCK activity in TGFβ-treated fibroblasts and sclera from ocular hypertensive mice was assessed by measuring phosphorylation of the ROCK substrate MYPT1 at Thr696. Fibroblast proliferation following IOP elevation and ROCK inhibitor treatment was assessed by an enzyme-linked immunosorbent (ELISA) assay. Results ROCK inhibitors H1152 (10μM), Y27632 (10 μM), and fasudil (5μM) reduced SMA expression 72%, 85%, and 68%, respectively. Collagen gel contraction was reduced by 36% (P < 0.001), 27% (P = 0.0003), and 33% (P = 0.0019) following treatment with fasudil (25 μM), Y27632 (10 μM), and H1152 (10μM). ROCK activity induced by TGFβ rose 4.74 ± 1.9 times over control at 4 hours (P = 0.0004) and 2.4 ± 0.47-fold (P = 0.0016) in sclera after IOP elevation. Proliferation of scleral fibroblasts after chronic IOP elevation was reduced 77% by Y27632 (P = 0.001) and 84% by fasudil (P = 0.0049). Conclusions ROCK inhibitors reduce TGFβ-induced myofibroblast transdifferentiation and glaucoma-induced scleral cell proliferation. Translational Relevance These findings suggest altered fibroblast activity promoted by ROCK inhibitors could modify scleral biomechanics and be relevant to glaucoma treatment.

Factors Influencing Central Lamina Cribrosa Depth: A Multicenter Study.

Abstract: Purpose:To quantify the influence of ocular and demographic factors on central laminar depth (LD) in healthy participants. Methods:A total of 362 normal subjects underwent optical coherence tomography (OCT) enhanced depth imaging of the optic nerve head (ONH) with a 24 radial B-scan pattern aligned to the fovea-to-Bruch's membrane opening (BMO) axis. BMO, anterior lamina, anterior scleral canal opening (ASCO), Bruch's membrane (BM), and the peripapillary scleral surface were manually segmented. The extent of laminar segmentation was quantified within 72 ASCO subsectors. Central LD was quantified relative to four reference planes: BMO, ASCO, BM, and scleral. The effects of age, sex, ethnicity, IOP, BMO area, ASCO area, and axial length on LD were assessed. Results:Laminar visibility was most consistent within the central ASCO (median 89%, range, 69%-95%). LDBMO and LDBM were significantly shallower in eyes with greater age, BMO area, and axial length and in females. LDASCO was shallower in eyes with greater ASCO area and axial length and in European and Hispanic descent compared to African descent eyes. LDSclera behaved similarly, but was not associated with axial length. BMO and ASCO area were not different between African descent and European descent eyes. Conclusions:Central LD was deeper in African descent eyes and influenced least by age, axial length, and sex, but more by ASCO area, when measured relative to the ASCO and sclera. However, the magnitude of these effects for all four reference planes was small, and their clinical importance in the detection of glaucoma and its progression remains to be determined.

Use of Animal Models and Techniques in Glaucoma Research: Introduction.

Abstract: This chapter provides an overview over the use of animal models in glaucoma research from a historical perspective. Strengths and limitations of various models are considered, and the challenges of translating results in animal experiments to successful clinical trials in humans are discussed.

Improved Outcomes for Transscleral Cyclophotocoagulation Through Optimized Treatment Parameters.

Abstract: PURPOSE To compare outcomes of transscleral diode cyclophotocoagulation with the treatment parameters used. PATIENTS AND METHODS This was a retrospective chart review of a random, 50% sample of diode procedures using the G-probe over 10 years for uncontrolled glaucoma. The main outcome measure was intraocular pressure (IOP) reduction by 20% and final IOP≤21 mm Hg. RESULTS In 236 eyes (persons) treated by 5 glaucoma specialists, most eyes had severe glaucoma, with 75% having <20/200 acuity. Median follow-up was 2.7 years. In eyes receiving only 1 treatment, IOP success criterion was met in 72% (129/180). Success was significantly related to power per delivery and median total joules per treatment (successes=135 J, failures=98 J; P=0.0009), but not to number of deliveries, nor to extent of circumference treated. Greater success was associated with 3 or 4 second duration per delivery, power level based on audible cues, and firm pressure on the sclera. Using a standard 2000 mW, 2 seconds, 20 deliveries in each eye had the lowest success (49%). Of those with no preoperative pain, 40 persons (57%) had no postoperative pain, whereas 20 reported pain of 1-3/10 (29%). Phthisis occurred in 7 eyes (3%), 5 of which had severe secondary eye disease. Nine eyes had no light perception preoperatively, whereas 50 eyes were no light perception at last follow-up, many after additional surgeries for other conditions. CONCLUSIONS Diode cyclophotocoagulation achieved reasonable IOP lowering, often without severe postoperative pain or complication. Greater success was achieved when audible effects were used to tailor the power settings to individual responses. Diode treatments with no intraoperative effect adjustment or using standardized protocols may not achieve optimal success.

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

Abstract: 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.

Age-Related Changes in Quantitative Strain of Mouse Astrocytic Lamina Cribrosa and Peripapillary Sclera Using Confocal Microscopy in an Explant Model.

Abstract: Purpose The purpose of this study was to measure the full-field deformation response to IOP change in the peripapillary sclera (PPS) and astrocytic lamina cribrosa (ALC) of young and old mouse eyes ex vivo. Methods Thirty-eight transgenic reporter mice with green fluorescent protein-expressing astrocytes were studied at 2 to 4 months and 13 to 15 months old. The ALC and PPS of the explant eyes were imaged using laser scanning microscopy under controlled inflation from 10 to 30 mm Hg. Strains were estimated for the ALC and PPS from imaged volumes using digital volume correlation. Results ALC strains were significantly greater than zero nasal-temporally for both age groups (mean = 4.3% and 4.0%; each P ≤ 0.004) and significantly greater than zero in the inferior-superior direction for younger mice (P = 0.0004). Younger mice had larger ALC inferior-superior strains than older mice (P = 0.002). The ALC area and perimeter enlarged with inflation in both age groups, with a greater increase in younger than in older mice (all P ≤ 0.004). The ALC nasal-temporal diameter change was greater than inferior-superiorly, and younger mice had greater enlargement nasal-temporally than older. PPS maximum shear strain was greater in the older mice (P = 0.002). The axial lengths of older mice were 14% longer and the PPS was 16% thinner than younger mice (both P = 0.0003). Conclusions The behavior of the ALC in younger mice with inflation exhibited greater strains and enlargement of ALC area than older mice. Some strain measures in the PPS were greater in older mice, likely related to their longer axial length and thinner PPS.

Sustained Dorzolamide Release Prevents Axonal and Retinal Ganglion Cell Loss in a Rat Model of IOP-Glaucoma.

Abstract: Purpose To determine if one injection of a sustained release formulation of dorzolamide in biodegradable microparticles (DPP) reduces retinal ganglion cell (RGC) loss in a rat model of glaucoma. Methods We injected either DPP or control microparticles intravitreally in rats. Two days later, unilateral ocular hypertension was induced by translimbal, diode laser treatment by a surgeon masked to treatment group. IOP and clinical exams were performed until sacrifice 6 weeks after laser treatment. RGC loss was measured by masked observers in both optic nerve cross-sections and RGC layer counts from retinal whole mounts. Results Cumulative IOP exposure was significantly reduced by DPP injection (49 ± 48 mm Hg × days in treated versus 227 ± 191 mm Hg × days in control microparticle eyes; P = 0.012, t-test). While control-injected eyes increased in axial length by 2.4 ± 1.7%, DPP eyes did not significantly enlarge (0.3 ± 2.2%, difference from control, P = 0.03, t-test). RGC loss was significantly less in DPP eyes compared with control microparticle injection alone (RGC axon count reduction: 21% vs. 52%; RGC body reduction: 25% vs. 50% [beta tubulin labeling]; P = 0.02, t-test). Conclusions A single injection of sustained release DPP protected against RGC loss and axial elongation in a rat model of IOP glaucoma. Translational relevance Sustained release IOP-lowering medications have the potential to stop glaucoma progression.

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

Abstract: 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.

Protruded retinal layers within the optic nerve head neuroretinal rim.

Abstract: PURPOSE To determine the frequency with which retinal tissues other than the nerve fibre layer, hereafter referred to as protruded retinal layers (PRL), are a component of optical coherence tomography (OCT) neuroretinal rim measurements. METHODS Ninety healthy (30 White, Black and Japanese, respectively) subjects were included in the study. A radial scan pattern (24 B-scans centred on Bruch's membrane opening [BMO]) was used. For each of the 48 minimum rim width (MRW) measurement points, we determined whether PRL were present, absent or indeterminate. When present, the proportion of PRL within the MRW was quantified. RESULTS Protruded retinal layers were present in 503 (11.6%), absent in 3805 (88.1%) and indeterminate in 12 (0.3%) measurement points. Overall, 69 (76.6%) subjects had ≥1 points with PRL, with White subjects having the highest frequency and Japanese the lowest (29 [97%] and 18 [60%], respectively; p < 0.01). PRL were present in one-third of points in the temporal sector, but ≤5% in other sectors. When present, the median PRL thickness was 53.0 (interquartile range [IQR]: 33.0 to 78.5) μm, representing 20.6 (IQR: 13.0 to 28.5)% of MRW. Globally, the median PRL thickness comprised 1.3 (IQR: 0.2 to 3.5)% of the MRW; however, in the temporal sector, it exceeded 30% of MRW in some subjects. CONCLUSIONS Protruded retinal layers are a component of MRW measurements in most normal subjects, occurring in almost 12% of all measurement points analysed. There were racial variations in the presence of PRL and a significantly higher frequency of PRL in the temporal sector.