Retinal Macroglial Responses in Health and Disease

Due to their permanent and close proximity to neurons, glial cells perform essential tasks for the normal physiology of the retina. Astrocytes andM¨uller cells (retinal macroglia) provide physical support to neurons and supplement them with several metabolites and growth factors.Macroglia are involved in maintaining the homeostasis of extracellular ions and neurotransmitters, are essential for information processing in neural circuits, participate in retinal glucose metabolism and in removing metabolic waste products, regulate local blood flow, induce the blood-retinal barrier (BRB), play fundamental roles in local immune response, and protect neurons from oxidative damage. In response to polyetiological insults, glia cells react with a process called reactive gliosis, seeking to maintain retinal homeostasis. When malfunctioning, macroglial cells can become primary pathogenic elements. A reactive gliosis has been described in different retinal pathologies, including age-related macular degeneration (AMD), diabetes, glaucoma, retinal detachment, or retinitis pigmentosa. A better understanding of the dual, neuroprotective, or cytotoxic effect of macroglial involvement in retinal pathologies would help in treating the physiopathology of these diseases.The extensive participation of the macroglia in retinal diseases points to these cells as innovative targets for new drug therapies.

Automatic Counting of Microglial Cells in Healthy and Glaucomatous Mouse Retinas

Proliferation of microglial cells has been considered a sign of glial activation and a hallmark of ongoing neurodegenerative diseases. Microglia activation is analyzed in animal models of different eye diseases. Numerous retinal samples are required for each of these studies to obtain relevant data of statistical significance. Because manual quantification of microglial cells is time consuming, the aim of this study was develop an algorithm for automatic identification of retinal microglia. Two groups of adult male Swiss mice were used: age-matched controls (naïve, n = 6) and mice subjected to unilateral laser-induced ocular hypertension (lasered; n = 9). In the latter group, both hypertensive eyes and contralateral untreated retinas were analyzed. Retinal whole mounts were immunostained with anti Iba-1 for detecting microglial cell populations. A new algorithm was developed in MATLAB for microglial quantification; it enabled the quantification of microglial cells in the inner and outer plexiform layers and evaluates the area of the retina occupied by Iba-1+ microglia in the nerve fiber-ganglion cell layer. The automatic method was applied to a set of 6,000 images. To validate the algorithm, mouse retinas were evaluated both manually and computationally; the program correctly assessed the number of cells (Pearson correlation R = 0.94 and R = 0.98 for the inner and outer plexiform layers respectively). Statistically significant differences in glial cell number were found between naïve, lasered eyes and contralateral eyes (P<0.05, naïve versus contralateral eyes; P<0.001, naïve versus lasered eyes and contralateral versus lasered eyes). The algorithm developed is a reliable and fast tool that can evaluate the number of microglial cells in naïve mouse retinas and in retinas exhibiting proliferation. The implementation of this new automatic method can enable faster quantification of microglial cells in retinal pathologies.

Effect of pharmacological pupil dilation on measurements and iol power calculation made using the new swept-source optical coherence tomography-based optical biometer

Purpose: to determine whether pupil dilation affects biometric measurements and intraocular lens (IOL) power calculation made using the new swept-source optical coherence tomography-based optical biometer (IOLMaster 700©; Carl Zeiss Meditec, Jena, Germany). Procedures: eighty-one eyes of 81 patients evaluated for cataract surgery were prospectively examined using the IOLMaster 700© before and after pupil dilation with tropicamide 1%. The measurements made were: axial length (AL), central corneal thickness (CCT), aqueous chamber depth (ACD), lens thickness (LT), mean keratometry (MK), white-to-white distance (WTW) and pupil diameter (PD). Holladay II and SRK/T formulas were used to calculate IOL power. Agreement between measurement modes (with and without dilation) was assessed through intraclass correlation coefficients (ICC) and Bland-Altman plots. Results: mean patient age was 75.17 ± 7.54 years (range: 57–92). Of the variables determined, CCT, ACD, LT and WTW varied significantly according to pupil dilation. Excellent intraobserver correlation was observed between measurements made before and after pupil dilation. Mean IOL power calculation using the Holladay 2 and SRK/T formulas were unmodified by pupil dilation. Conclusions: the use of pupil dilation produces statistical yet not clinically significant differences in some IOLMaster 700© measurements. However, it does not affect mean IOL power calculation.

Cell Pattern in Adult Human Corneal Endothelium

A review of the current data on the cell density of normal adult human endothelial cells was carried out in order to establish some common parameters appearing in the different considered populations. From the analysis of cell growth patterns, it is inferred that the cell aging rate is similar for each of the different considered populations. Also, the morphology, the cell distribution and the tendency to hexagonallity are studied. The results are consistent with the hypothesis that this phenomenon is analogous with cell behavior in other structures such as dry foams and grains in polycrystalline materials. Therefore, its driving force may be controlled by the surface tension and the mobility of the boundaries.

Agreement and clinical comparison between a new swept-source optical coherence tomography-based optical biometer and an optical low-coherence reflectometry biometer

Purpose To compare measurements taken using a swept-source optical coherence tomography-based optical biometer (IOLmaster 700) and an optical low-coherence reflectometry biometer (Lenstar 900), and to determine the clinical impacts of differences in their measurements on intraocular lens (IOL) power predictions. Methods Eighty eyes of 80 patients scheduled to undergo cataract surgery were examined with both biometers. The measurements made using each device were axial length (AL), central corneal thickness (CCT), aqueous depth (AQD), lens thickness (LT), mean keratometry (MK), white-to-white distance (WTW), and pupil diameter (PD). Holladay 2 and SRK/T formulas were used to calculate IOL power. Differences in measurement between the two biometers were determined using the paired t-test. Agreement was assessed through intraclass correlation coefficients (ICC) and Bland–Altman plots. Results Mean patient age was 76.3±6.8 years (range 59–89). Using the Lenstar, AL and PD could not be measured in 12.5 and 5.25% of eyes, respectively, while IOLMaster 700 took all measurements in all eyes. The variables CCT, AQD, LT, and MK varied significantly between the two biometers. According to ICCs, correlation between measurements made with both devices was excellent except for WTW and PD. Using the SRK/T formula, IOL power prediction based on the data from the two devices were statistically different, but differences were not clinically significant. Conclusions No clinically relevant differences were detected between the biometers in terms of their measurements and IOL power predictions. Using the IOLMaster 700, it was easier to obtain biometric measurements in eyes with less transparent ocular media or longer AL.

Melatonin Makes Me Feel Awake! Seven Years of Lab Experience (2000-2007)

The research on the effect of melatonin on intraocular pressure (IOP) is reviewed from the hystorical point of view of our laboratory. The original idea of melatonin modulating intraocular pressure has been improved by using selective compounds for MT2 and specially melatonin MT3 receptors. The selective compound 5-methoxyamino N-acetyltryptamine (5-MCA-NAT) has been an attractive compound due to its ability to reduce IOP about 40%, therefore being a good candidate to the treatment of the ocular hypertension linked to glaucoma. More compounds have been developed and tested permitting us to have a more accurate panorama of those receptors controlling the relevant process of intraocular pressure.

A Diagnostic Calculator for Detecting Glaucoma on the Basis of Retinal Nerve Fiber Layer, Optic Disc, and Retinal Ganglion Cell Analysis by Optical Coherence Tomography

Purpose: The purpose of this study was to develop and validate a multivariate predictive model to detect glaucoma by using a combination of retinal nerve fiber layer (RNFL), retinal ganglion cell-inner plexiform (GCIPL), and optic disc parameters measured using spectral-domain optical coherence tomography (OCT). Methods: Five hundred eyes from 500 participants and 187 eyes of another 187 participants were included in the study and validation groups, respectively. Patients with glaucoma were classified in five groups based on visual field damage. Sensitivity and specificity of all glaucoma OCT parameters were analyzed. Receiver operating characteristic curves (ROC) and areas under the ROC (AUC) were compared. Three predictive multivariate models (quantitative, qualitative, and combined) that used a combination of the best OCT parameters were constructed. A diagnostic calculator was created using the combined multivariate model. Results: The best AUC parameters were: inferior RNFL, average RNFL, vertical cup/disc ratio, minimal GCIPL, and inferior-temporal GCIPL. Comparisons among the parameters did not show that the GCIPL parameters were better than those of the RNFL in early and advanced glaucoma. The highest AUC was in the combined predictive model (0.937; 95% confidence interval, 0.911–0.957) and was significantly (P = 0.0001) higher than the other isolated parameters considered in early and advanced glaucoma. The validation group displayed similar results to those of the study group. Conclusions: Best GCIPL, RNFL, and optic disc parameters showed a similar ability to detect glaucoma. The combined predictive formula improved the glaucoma detection compared to the best isolated parameters evaluated. The diagnostic calculator obtained good classification from participants in both the study and validation groups.

Macroglial and retinal changes in hypercholesterolemic rabbits after normalization of cholesterol levels

This study evaluates hypercholesterolemic rabbits, examining the retinal changes in Müller cells and astrocytes as well as their variations after a period of normal blood-cholesterol values induced by a standard diet. New Zealand rabbits were divided into three groups: G0, fed a standard diet; G1A, fed a 0.5% cholesterol-enriched diet for 8 months; and G1B, fed as G1A followed by standard diet for 6 months. Eyes were processed for transmission electron microscopy and immunohistochemistry (GFAP). While G1B resembled G0 more than did G1A, they shared alterations with G1A: a) as in G1A, Müller cells were GFAP+, filled spaces left by axonal degeneration, formed glial scars and their nuclei were displaced to the nerve-fibre layer. The area occupied by the astrocytes associated with the nerve-fibre bundles (AANFB) and by perivascular astrocytes (PVA) in G1A and G1B was significantly lower than in controls. However, no significant differences in PVA were found between G1A and G1B. In G1B, type I PVA was absent and replaced by hypertrophic type II cells; b) Bruch's membrane (BM) was thinner in G1B than in G1A; c) the retinal pigment epithelium (RPE) cytoplasm contained fewer lipids in G1B than in G1A; d) in G1A and G1B choriocapillaris and retinal vessel showed alterations with respect to G0; e) cell death and axonal degeneration in the retina were similar in G1A and G1B. The substitution of a hyperlipemic diet by a standard one normalizes blood-lipid levels. However, the persistence of damage at retinal vessels and BM-RPE could trigger chronic ischemia.

Tratamiento y seguimiento de la degeneración macular asociada a la edad (DMAE)

En la actualidad la degeneración macular asociada a la edad (DMAE) es una de las patologías más importantes debido a que es la principal causa de ceguera en los mayores de 50 años y por tanto produce una gran incapacidad. Factores como la edad, tabaco, ejercicio físico, aumentan la probabilidad de que se padezca DMAE; entre estos factores cabe destacar la edad ya que debido al aumento de la esperanza de vida su prevalencia es cada vez mayor, afectando tanto a la calidad de vida del paciente como a la independencia. El tratamiento de dicha enfermedad depende del tipo que se padezca, DMAE húmeda o seca, y aunque en ningún caso va a curar la enfermedad se puede paralizar y llegar a mejorar la calidad de vida de los pacientes. El tratamiento con mejores resultados es el correspondiente al uso de los inhibidores de la angiogénesis, aunque existen otros tratamientos como son la terapia fotodinámica, la fotocoagulación con láser, la ozonoterapia, o el tratamiento con Dobesilato que es una de las últimas novedades descubiertas para el tratamiento de la DMAE. Es de vital importancia llevar a cabo un seguimiento adecuado a los pacientes para ver cómo evoluciona la enfermedad y controlarla.

Visual outcomes after bilateral trifocal diffractive intraocular lens implantation

Background In recent years new models of intraocular lenses are appearing on the market to reduce requirements for additional optical correction. The purpose of this study is to assess visual outcomes following bilateral cataract surgery and the implant of a FineVision® trifocal intraocular lens (IOL). Methods Prospective, nonrandomized, observational study. Vision was assessed in 44 eyes of 22 patients (mean age 68.4 ± 5.5 years) before and 3 months after surgery. Aberrations were determined using the Topcon KR-1 W wave-front analyzer. LogMAR visual acuity was measured at distance (corrected distance visual acuity, CDVA 4 m), intermediate (distance corrected intermediate visual acuity, DCIVA 60 cm) and near (distance corrected near visual acuity, DCNVA 40 cm). The Pelli-Robson letter chart and the CSV-1000 test were used to estimate contrast sensitivity (CS). Defocus curve testing was performed in photopic and mesopic conditions. Adverse photic phenomena were assessed using the Halo v1.0 program. Results Mean aberration values for a mesopic pupil diameter were: total HOA RMS: 0.41 ± 0.30 μm, coma: 0.32 ± 0.22 μm and spherical aberration: 0.21 ± 0.20 μm. Binocular logMAR measurements were: CDVA −0.05 ± 0.05, DCIVA 0.15 ± 0.10, and DCNVA 0.06 ± 0.10. Mean Pelli-Robson CS was 1.40 ± 0.14 log units. Mean CSV100 CS for the 4 frequencies examined (A: 3 cycles/degree (cpd), B: 6 cpd, C: 12 cpd, D: 18 cpd) were 1.64 ± 0.14, 1.77 ± 0.18, 1.44 ± 0.24 and 0.98 ± 0.24 log units, respectively. Significant differences were observed in defocus curves for photopic and mesopic conditions (p < 0.0001). A mean disturbance index of 0.28 ± 0.22 was obtained. Conclusions Bilateral FineVision IOL implant achieved a full range of adequate vision, satisfactory contrast sensitivity, and a lack of significant adverse photic phenomena. Trial registration Eudract Clinical Trials Registry Number: 2014-003266-2.

TRPV4 activation triggers the release of melatonin from human non-pigmented ciliary epithelial cells

Melatonin is a neurohormone mainly produced in the pineal gland; nevertheless, various ocular structures such as the ciliary body, lens and the retina produce it. One of the roles of melatonin in the eye is the modulation of intraocular pressure, although little is known about the mechanisms that causes its presence in the aqueous humour. TRPV4 is a membrane channel which is activated by both physical and chemical stimuli. Therefore, this channel is sensitive to osmotic and hydrostatic pressure. As a consequence, TRPV4 results as an interesting candidate to study the relation between the activation of the TRPV4 channel and the production of melatonin. In this sense we have studied the role of the TRPV4 agonist GSK1016790A to modulate the production of melatonin in a cell line derived from human non-pigmented ciliary epithelial cells. The stimulation of the TRPV4 produced an increase in the extracellular melatonin levels changing from 8.5 ± 0.6 nM/well/30 min (control) to 23.3 ± 2.1 nM/well/30 min after 10 nM GSK1016790A application, this action being blocked by the selective antagonist RN 1734. The activation of the TRPV4 by GSK1016790A permitted to observe a melatonin increase which was concentration-dependent, and provided a pD2 value of −8.5 ± 0.1 (EC50 of 3.0 nM). In conclusion, the activation of the TRPV4 present in human non-pigmented ciliary epithelial cells can modulate the presence of extracellular melatonin, this being of relevance since this substance controls the dynamics of the aqueous humour.

Accommodative changes produced in response to overnight orthokeratology

Background To evaluate short-term (3 months) and long-term (3 years) accommodative changes produced by overnight orthokeratology (OK). Methods A prospective, longitudinal study on young adult subjects with low to moderate myopia was carried out. A total of 93 patients took part in the study. Out of these, 72 were enrolled into the short-term follow-up: 21 were on a control group, 26 on a Paragon CRT contact lenses group, and 25 on a Seefree contact lenses group. The other 21 patients were old CRT wearers on long-term follow-up. Accommodative function was assessed by means of negative and positive relative accommodation (NRA / PRA), monocular accommodative amplitude (MAA), accommodative lag, and monocular accommodative facility (MAF). These values were compared among the three short-term groups at the follow-up visit. The long- and short-term follow-up data was compared among the CRT groups. Results Subjective accommodative results did not suffer any statistically significant changes in any of the accommodative tests for any of the short-term groups when compared to baseline. There were no statistically significant differences between the three short-term groups at the follow-up visit. When comparing the short- and long-term groups, only the NRA showed a significant difference (p = 0.0006) among all the accommodation tests. Conclusions OK does not induce changes in the ocular accommodative function for either short-term or long-term periods.