Systemic epigallocatechin gallate protects against retinal degeneration and hepatic oxidative stress in the P23H-1 rat.

Retinitis pigmentosa (RP) is a group of inherited retinal disorders that lead to photoreceptor loss. RP has been reported to be related to oxidative stress, autophagy, and inflammation. (-)-Epigallocatechin gallate (EGCG), the most abundant catechin-based flavonoid in green tea leaves, has significant antioxidant, anti-carcinogenic, antimicrobial, and neuroprotective properties. EGCG, given its low molecular weight and hydrophilic properties, can cross the blood-retinal barrier and is able to reach different ocular tissues such as the lens, cornea, and retina. EGCG has been shown to provide retinal protection against ischemia; sodium nitroprusside-, N-methyl-D-aspartate-, lipopolysaccharide-, light-, sodium iodate-, or H2O2-induced damage and diabetic retinopathy. This suggests that systemic EGCG administration has the potential to protect against retinal degenerative or neurodegenerative diseases such as RP. The aim of this work was to investigate whether EGCG can protect against RP progression in the animal P23H line 1, the model of RP. Albino P23H rats were crossed with pigmented Long Evans rats to produce offspring exhibiting the clinical features of RP. Pigmented P23H rats were treated via intraperitoneal injection with saline or EGCG at a dose of 25 mg/kg every week from P100 to P160 and then compared to wild-type Long Evans rats. Rats treated with EGCG showed better visual and retinal electrical function with increased contrast sensitivity and b-wave values compared with those observed in P23H rats treated with vehicle. EGCG reduced lipid peroxidation and increased total antioxidant capacity and catalase and superoxide dismutase activities. No differences were observed in visual acuity, nitrate levels, nitrite levels or glutathione S-transferase activity. In conclusion, EGCG not only reduced the loss of visual function in P23H rats but also improved the levels of antioxidant enzymes and reduced oxidative damage. This study was approved by the Institutional Animal Care and Use Committee (CEICA) from the University of Zaragoza under project license PI12/14 on July 11, 2014.

Effects of Daily Melatonin Supplementation on Visual Loss, Circadian Rhythms, and Hepatic Oxidative Damage in a Rodent Model of Retinitis Pigmentosa.

Retinitis pigmentosa (RP) is a group of inherited neurodegenerative diseases characterized by a progressive loss of visual function that primarily affect photoreceptors, resulting in the complete disorganization and remodeling of the retina. Progression of the disease is enhanced by increased oxidative stress in the retina, aqueous humor, plasma, and liver of RP animal models and patients. Melatonin has beneficial effects against age-related macular degeneration, glaucoma, and diabetic retinopathy, in which oxidative stress plays a key role. In the present study, we used the P23HxLE rat as an animal model of RP. Melatonin treatment (10 mg/kg b.w. daily in drinking water for 6 months) improved the parameters of visual function and decreased the rate of desynchronization of the circadian rhythm, both in P23HxLE and wild-type rats. Melatonin reduced oxidative stress and increased antioxidant defenses in P23HxLE animals. In wild-type animals, melatonin did not modify any of the oxidative stress markers analyzed and reduced the levels of total antioxidant defenses. Treatment with melatonin improved visual function, circadian synchronization, and hepatic oxidative stress in P23HxLE rats, an RP model, and had beneficial effects against age-related visual damage in wild-type rats.

Changes in retinal layers in type 1 diabetes mellitus without retinopathy measured by spectral domain and swept source OCTs.

To evaluate changes in inner retinal layer (IRL) thicknesses in patients with type 1 diabetes mellitus (DM1) with no diabetic retinopathy (DR) using two different optical coherence tomography (OCT) devices. Ninety DM1 and 60 healthy eyes were evaluated using spectral domain (SD)-OCT and swept source (SS)-OCT to measure changes in the retinal nerve fiber layer (RNFL), ganglion cell layer (GCL), inner plexiform layer (IPL) and inner nuclear layer (INL) thicknesses in all Early Treatment of Diabetic Retinopathy Study (ETDRS) macular areas. Functional tests were performed in both groups, including ETDRS with 100, 2.5 and 1.25% contrast, and color vision. The mean ages were 42.93 ± 13.62 and 41.52 ± 13.05 years in the diabetic and control groups, respectively. Visual acuity (VA) with ETDRS 1.25% was lower in the DM1 patients. Both ETDRS 2.5% and color vision were lower in the DM1 group but did not reach statistical significance. Retinal thicknesses in the central area and in the vertical outer areas were higher in the DM1 group. Differences were found in the IRL with no changes in the outer ones. Long-term DM1 patients with no DR maintained visual function, with a decrease in VA with 1.25% ETDRS contrast. Macular thickness measurements were higher using Spectralis SD-OCT than DRI Triton SS-OCT, and DM1 patients had a decrease in IRL thickness, especially in the GCL at the parafoveal level, generating thinning of the RNFL in the peripheral areas. There were no differences in outer retinal layer (ORL) thickness.

Microperimetry and Optical Coherence Tomography Changes in Type-1 Diabetes Mellitus without Retinopathy.

BACKGROUND: We aimed to measure and correlate inner retinal layer (IRL) thickness and macular sensitivity by optical coherence tomography (OCT) and by microperimetry, respectively, in type 1 diabetes mellitus patients (DM1) without diabetic retinopathy (DR). METHODS: Fifty-one DM1 patients and 81 age-matched healthy subjects underwent measurement of the axial length (AL), retinal thickness in the macular ETDRS areas by swept source (SS)-OCT and macular sensitivity by microperimeter. RESULTS: The total retinal and IRL thicknesses were thicker in the DM1 group (p < 0.05) in practically all ETDRS areas, and they had a generalized decrease in sensitivity (p < 0.05) in 9 areas between both groups. There was a significant negative correlation between retinal sensitivity and age in all areas and in visual acuity (VA) in 5 out of the 9 areas for DM1 patients. Only a mild negative correlation was observed between retinal sensitivity in the 5° nasal inner (5NI) area and in IRL thickness in the temporal inner (TI) area (-0.309 with p = 0.029) in the DM1 group. CONCLUSION: Aging and disease evolution in DM1 patients without DR signs generate a decrease in retinal sensitivity. There was a direct relationship between retinal sensitivity and macular thickness in the DM1 group.

Choroidal Thickness and Volume Modifications Induced by Aerobic Exercise in Healthy Young Adults.

INTRODUCTION: Our aim was to evaluate the changes in choroidal thickness (CT) and volume (CV) following aerobic physical exercise in healthy young adults. METHODS: This study included 72 eyes from healthy volunteers between 22 and 37 years old. Using the International Physical Activity Questionnaire, total physical activity was computed. Measurements using an autorefractometer, ocular biometry, and spectral-domain optical coherence tomography using the Enhanced Depth Imaging protocol were taken. OCT was performed as a baseline measurement and after performing 10 min of dynamic physical exercise (3 and 10 min post-exercise). The choroidal layer was manually segmented, and the CT and CV in different areas from the Early Treatment Diabetic Retinopathy Study grid were obtained. RESULTS: In healthy adults, at 3 min post-exercise, CT was higher in the subfoveal, the 3-mm nasal, and the 6-mm superior areas. Between 3 and 10 min post-exercise, the CT was reduced in all areas, and in some areas, the values were even smaller than the baseline measurements. The CV values showed changes after exercise similar to those of thickness. The total CV recovery after exercise was related to sex and physical activity level. CONCLUSION: Individuals with higher physical activity habits had greater CV at rest than those with lower physical activity levels. During exercise, healthy young people adjust CT and CV. At 3 min post-exercise, CT and CV increase. Women and individuals with greater physical activity levels reduce their total CV more than others during recovery.

Choroidal Differences between Spectral and Swept-source Domain Technologies.

PURPOSE OF THE STUDY: To assess differences in healthy eyes in volume (CV) and choroidal thicknesses (CT) across all macular Early Treatment Diabetic Retinopathy Study (ETDRS) areas measured by swept source (SS)-OCT and spectral domain (SD)-OCT with enhanced depth imaging (EDI). MATERIALS AND METHODS: One hundred and fifty healthy eyes were studied in patients aged between 21-68 years. All of these patients underwent an ophthalmic examination that included axial length (AL), a fast macula protocol with EDI by Spectralis SD-OCT, and a 3D macular cube by DRI-Triton SS-OCT. CT and CV values were measured after a manual segmentation with SD-OCT and automatically with SS-OCT. RESULTS: The values obtained by SD-OCT were statistically higher than the values obtained by SS-OCT in all ETDRS areas. To determine the differences that were related to total CV or AL, the eyes were divided depending on their total CV and AL values into two groups. There were persistent differences between the two devices that were not related to the total CV. In subjects with a higher AL, these differences were not present in either the total CV or the subfoveal CT. In longer eyes, the differences diminished, and there were no statistical differences between the devices in the subfoveal area, temporal quadrants, inferior inner ring, or CV. CONCLUSIONS: The choroidal values obtained with the SD-OCT are statistically higher than those measured by SS-OCT. Both techniques are able to clearly visualize the choroidoscleral interface and provide different values that are highly correlated. These differences are smaller in eyes with a longer AL and are not related to the total CV.

Epigallocatechin Gallate Slows Retinal Degeneration, Reduces Oxidative Damage, and Modifies Circadian Rhythms in P23H Rats.

Retinitis pigmentosa (RP) includes a group of genetic disorders that involve the loss of visual function due to mutations mainly in photoreceptors but also in other retinal cells. Apoptosis, retinal disorganization, and inflammation are common in the progression of the disease. Epigallocatechin gallate (EGCG) has been proved as beneficial in different eye diseases. Pigmented heterozygous P23H rat was used as an animal model of RP. Visual function was assessed by optomotor and electroretinogram (ERG) and circadian rhythms were evaluated by telemetry. Hepatic oxidative damage and antioxidant defenses were assessed using biochemical tests. The visual function of the EGCG P23H group was preserved, with a deterioration in the activity period and lower values in the interdaily stability parameter. Control rats treated with EGCG were less active than the sham group. EGCG increased antioxidant levels in P23H rats but reduced total hepatic antioxidant capacity by almost 42% in control rats compared to the sham group. We conclude that treatment with EGCG improves visual function and antioxidant status in P23H rats but diminishes antioxidant defenses in wild-type control animals, and slightly worsens activity circadian rhythms. Further studies are necessary to clarify the beneficial effects in disease conditions and in healthy organisms.

Measurement method of optical properties of ex vivo biological tissues of rats in the near-infrared range.

An optical fiber-based supercontinuum setup and a custom-made spectrophotometer that can measure spectra from 1100 to 2300 nm, are used to describe attenuation properties from different ex vivo rat tissues. Our method is able to differentiate between scattering and absorption coefficients in biological tissues. Theoretical assumptions combined with experimental measurements demonstrate that, in this infrared range, tissue attenuation and absorption can be accurately measured, and scattering can be described as the difference between both magnitudes. Attenuation, absorption, and scattering spectral coefficients of heart, brain, spleen, retina, and kidney are given by applying these theoretical and experimental methods. Light through these tissues is affected by high scattering, resulting in multiple absorption events, and longer wavelengths should be used to obtain lower attenuation values. It can be observed that the absorption coefficient has a similar behavior in the samples under study, with two main zones of absorption due to the water absorption bands at 1450 and 1950 nm, and with different absolute absorption values depending on the constituents of each tissue. The scattering coefficient can be determined, showing slight differences between retina and brain samples, and among heart, spleen and kidney tissues.

Choroidal Changes of Long-Term Type 1 Diabetic Patients without Retinopathy.

The aim of the study is to assess choroidal thickness (CT) and choroidal volume (CV) in 90 type 1 diabetes mellitus (DM1) patients with no diabetic retinopathy (DR) and 60 control eyes using spectral domain optical coherence tomography (SD-OCT) and swept source (SS)-OCT in the areas of the Early Treatment Diabetic Retinopathy Study (ETDRS). Mean ages were 42.93 ± 13.62 and 41.52 ± 13.05 years in the diabetic and control groups, respectively. Significant differences were obtained between both groups with Spectralis SD-OCT in all ETDRS areas and in the total CV, excluding the temporal perifoveal one. With Triton SS-OCT, statistically significant differences were obtained in the subfoveal CT and in the vertical areas. CT showed the same tendency with both OCTs, with greater CT and CV in the DM1 group than the mean values of the control group. To assess the influence of DM1 evolution in the CT modifications, DM1 patients were divided into Group 1, with less than 24 years of diagnosis, and Group 2, with ≥24 years of DM1 evolution. Using both OCTs, seven of the nine ETDRS areas and the CV had lower values in Group 2. CT and CV measured by OCT were higher in DM1 without DR. There is a choroidal thinning related to disease evolution in DM1. In patients with DM evolution greater than 24 years, the CT is statistically lower than in patients with less evolution of the disease.

CHANGES IN TOTAL AND INNER RETINAL THICKNESSES IN TYPE 1 DIABETES WITH NO RETINOPATHY AFTER 8 YEARS OF FOLLOW-UP.

PURPOSE: To evaluate changes in retinal layer thickness in patients with Type 1 diabetes with no diabetic retinopathy after 8 years of follow-up. METHODS: Ninety Type 1 diabetes and 60 control eyes were studied. Changes in the retinal nerve fiber layer, ganglion cell layer, and inner nuclear layer thicknesses in all Early Treatment Diabetic Retinopathy Study areas were evaluated. RESULTS: The mean ages were 42.93 ± 13.62 and 41.52 ± 13.05 years in the diabetic and control group, respectively. In 2009, total retinal thickness was higher in diabetic patients; differences were statistically significant in all except the nasal areas. In both groups, the mean foveal thickness remained the same during the 8 years. Among diabetic patients, there was a significant reduction in total retinal thickness in all areas excluding the outer temporal one; controls only in the inferior areas. The thickness loss was due to the thinning of the inner retinal layers (inner nuclear layer, ganglion cell layer, and retinal nerve fiber layer). The controls showed a significant diminution in the retinal nerve fiber layer and in the ganglion cell layer areas. The inner nuclear layer showed a diminution in the diabetes mellitus group. CONCLUSION: Before the onset of diabetic retinopathy, Type 1 diabetes patients experience a diminution of their inner retinal layer thicknesses over time, supporting the hypothesis of retinal neurodegeneration.

Hepatic oxidative stress in pigmented P23H rhodopsin transgenic rats with progressive retinal degeneration.

Retinitis pigmentosa (RP) comprises a group of inherited retinal degenerative conditions characterized by primary degeneration of the rod photoreceptors. Increased oxidative damage is observed in the retina, aqueous humor, and plasma of RP animal models and patients. The hepatic oxidative status may also be affected in RP due to oxidative damage influencing soluble macromolecules exiting the retina or to alterations in the melanopsin system resulting in chronic circadian desynchronization that negatively alters the oxidative stress defense system. P23H rats were crossed with pigmented Long Evans rats to produce offspring exhibiting the clinical conditions of RP. We measured hepatic malondialdehyde and 4-hydroxyalkenal concentrations as oxidative stress markers; nitrite level as a total nitrosative damage marker; total antioxidant capacity; and the activities of catalase, superoxide dismutase (SOD), and glutathione S-transferase. Retinal visual function was assessed based on optomotor and electroretinogram responses. P23H transgenic rats exhibited diminished visual acuity, contrast sensitivity, and electroretinographic responses according to the level of retinal degeneration. P23H rats at 30 days of age already demonstrated only 47% of the hepatic total antioxidant capacity of wild-type animals. Hepatic catalase and SOD activities were also reduced in P23H rats after 120 days, but we detected no difference in glutathione S-transferase activity. P23H rats had increased hepatic oxidative and nitrosative damage markers. GSH/GSSG ratio showed a significant diminution in P23H rats at P120 compared to WT. We conclude that the liver is under increased oxidative stress in P23H rats. Further studies are required, however, to clarify the contribution of systemic oxidative damage to the pathogenesis of RP.

Development of optokinetic tracking software for objective evaluation of visual function in rodents.

The aim of this study was to develop software that performs the optokinetic tracking assessment without the involvement of experimenters to increase the objectivity of the test. To check the effectiveness of the software, several videos were analyzed and the results were compared to those produced by two experimenters. Videos consisted of visual acuity and contrast sensitivity tests on normal animals and pigmented P23H rats (animal model of retinitis pigmentosa). Our software showed a reasonably high success rate: in approximately 78% of the cases, both the software program and the experimenters were in agreement, including the direction of rotation. The software detected 7% false positive cases, 10% false negative cases, and it was wrong in 5% of the cases. Decrease in visual function with age in pigmented P23H rats was observed from the first time interval, although minimum thresholds were found in visual parameters at advanced ages. We developed simple software based on current functions included in the Matlab image processing toolbox that was able to recognize, with a reasonably high percentage of success, the subtle head movements of the rodent produced when visual perception of the optokinetic optotype occurs.

Spectral attenuation of brain and retina tissues in the near-infrared range measured using a fiber-based supercontinuum device.

A novel setup for the efficient constant optical measurements of biological tissues in the near infrared is presented. The system combines the use of a fiber-based supercontinuum source with a simple optics fiber collimator. This configuration allows a wide spectral range of measurement and, at the same time, can efficiently filter the straightforward transmitted light while avoiding scattered light. As a performance example, the optical characterization of rat brain and retina tissues are shown. The attenuation coefficient for both tissues in the near infrared region is also obtained. This technique could be applied in clinical research as a noninvasive method with several potential practical applications.