Various cross-linking methods inhibit the collagenase I degradation of rabbit scleral tissue.

BACKGROUND: Collagen cross-linking of the sclera is a promising approach to strengthen scleral rigidity and thus to inhibit eye growth in progressive myopia. Additionally, cross-linking might inhibit degrading processes in idiopathic melting or in ocular inflammatory diseases of the sclera. Different cross-linking treatments were tested to increase resistance to enzymatic degradation of the rabbit sclera. METHODS: Scleral patches from rabbit eyes were cross-linked using paraformaldehyde, glutaraldehyde or riboflavin combined with UV-A-light or with blue light. The patches were incubated with collagenase I (MMP1) for various durations up to 24 h to elucidate differences in scleral resistance to enzymatic degradation. Degraded protein components in the supernatant were detected and quantified using measurements of Fluoraldehyde o-Phthaldialdehyde (OPA) fluorescence. RESULTS: All cross-linking methods reduced the enzymatic degradation of rabbit scleral tissue by MMP1. Incubation with glutaraldehyde (1%) and paraformaldehyde (4%) caused nearly a complete inhibition of enzymatic degradation (down to 7% ± 2.8 of digested protein compared to control). Cross-linking with riboflavin/UV-A-light reduced the degradation by MMP1 to 62% ± 12.7 after 24 h. Cross-linking with riboflavin/blue light reduced the degradation by MMP1 to 77% ± 13.5 after 24 h. No significant differences could be detected comparing different light intensities, light exposure times or riboflavin concentrations. CONCLUSIONS: The application of all cross-linking methods increased the resistance of rabbit scleral tissue to MMP1-degradation. Especially, gentle cross-linking with riboflavin and UV-A or blue light might be a clinical approach in future.

Analysis of foveal characteristics and their asymmetries in the normal population.

The advance of optical coherence tomography (OCT) enables a detailed examination of the human retina in-vivo for clinical routine and experimental eye research. Only few investigations to date captured human foveal morphology in a large subject group on the basis of a detailed analysis employing mathematical models. However, even for important foveal characteristics unified terminology and clear definitions were not implemented so far. This might be a reason, why to this day the human fovea is considered to be a mostly symmetric and round structure. Therefore, the most important finding of this work is the detailed analysis of the asymmetric structure of the human fovea. We employed five clinically highly relevant foveal characteristics, which are derived from a previously published fovea model. For each, an accurate mathematical description is given. The presented properties include (1) mean retinal thickness inside a defined radius, (2) foveal bowl area, (3) a new, exact definition of foveal radius, (4) maximum foveal slope, and (5) the maximum height of the foveal rim. Furthermore, minimum retinal thickness was derived and analyzed. 220 strictly controlled healthy Caucasian subjects of European decent with an even distribution of age and gender were imaged with an Heidelberg Spectralis OCT. Detailed analysis demonstrated the following general results: (1) significant gender difference regarding the central foveal subfield thickness (CFST) but no significant differences for the minimum central retinal thickness, (2) a strong correlation between right and left eye of the same subject, and, as essential finding, (3) strong structural differences of the fovea form in the different anatomical directions (nasal, temporal, inferior and superior). In the analysis of the foveal asymmetry, it will be demonstrated that the foveal radius is larger in nasal and temporal direction compared to inferior and superior position. Furthermore, it will be shown that the circular fovea rather has an elliptic form with the larger axis along the nasal to temporal direction. Interestingly, the foveal slope shows a divergent behavior as the temporal direction has the smallest slope angle and both, inferior and superior angles are clearly larger than the others. The findings in this work can be used for an exact quantification of changes in early stages of various retinal diseases and as a marker for initial diagnosis.

Influence of macular oedema on the measurement of macular pigment optical density.

INTRODUCTION: The purpose of this study was to determine macular pigment optical density (MPOD) in patients with macular degeneration as well as in patients with non-proliferative diabetic retinopathy. METHODS: Fifty-one phakic patients with either age-related macular degeneration (60 eyes of 30 patients; average age, 70.9 years) or non-proliferative diabetic retinopathy (42 eyes of 21 patients; average age, 61.7 years) were included in this cross-sectional study. Within the groups, patients were divided into those suffering from macular oedema and those with no oedema. An intra-subject comparison between eyes was carried out in both groups. Data were investigated on the basis of the coefficient of determination (R (2)). Macular pigment optical density was measured by fundus reflectometry using the one-wavelength reflection method (Visucam 500; Carl Zeiss Meditec AG, Jena, Germany), in conformity with the method described by Schweitzer et al. (2010). We evaluated the maximum optical density in the measurement area (max OD) and the average optical density across the reference area in the measurement area (mean OD). Specifically, the influence of macular oedema on macular pigment optical density was examined. The subsequent measurement of retinal thickness was carried out by spectral-domain optical coherence tomography (Spectralis SD-OCT, Heidelberg Engineering GmbH, Heidelberg, Germany). RESULTS: The current study included two groups. The first group consisted of patients with non-proliferative diabetic retinopathy, as follows: no macular oedema on either side (max OD: R (2) = 43.2 %, p = 0.16; mean OD: R (2) = 68.7 %, p = 0.04); one-sided macular oedema (max OD: R(2) = 16 %, p = 0.60; mean OD: R(2) = 100 %, p = 0.04); or macular oedema in both eyes (max OD: R(2) = 79.7 %, p < 0.01; mean OD: R(2) = 81.4 %, p < 0.01). The second group comprised patients with age-related macular degeneration (AMD), as follows: non-exudative changes on both sides (max OD: R(2) = 64.0 %, p = 0.20; mean OD: R (2) = 16 %, p = 0.60); one-sided exudative macular changes (max OD: R (2) = 50.6 %, p < 0.01; mean OD: R (2) = 20.8 %, p = 0.04); or exudative macular degeneration on both sides (max OD: 3 R (2) = 6.0 %, p = 0.29; mean OD: R (2)= 81.0 %, p = 0.04). The data available presented a correlation of MPOD values of both eyes of an individual within the groups investigated. In this respect, the data of the partner eyes within the group of patients with diabetic retinopathy were more highly correlated with each other than the values of both eyes of patients suffering from age-related macular degeneration. CONCLUSIONS: The present study showed that macular oedema did not seem to have an influence on a valid measurement of MPOD by one-wavelength fundus reflectometry. Thus, meaningful data could also be obtained on patients with exudative retinal changes.

Macular pigment optical density measurements by one-wavelength reflection photometry--influence of cataract surgery on the measurement results.

PURPOSE: The main objective of the present study was the investigation of possible influence of lens opacification on macular pigment optical density (MPOD) measurements. METHODS: Eighty-six eyes of 64 patients (mean age 73.4 ± 8.3 years) were included in the study. MPOD was prospectively measured using the one-wavelength reflection method (Visucam500, Carl Zeiss Meditec AG) before and after cataract extraction, with implantation of a blue-light filtering intraocular lens (AlconSN60WF). The median of the maximum optical density (MaxOD) and the median of the mean optical density (MeanOD) measurements of macular pigment across the subject group were evaluated. RESULTS: Statistically significant differences were noticed between pre-operative and post-operative measurements, the absolute values were generally lower after cataract extraction. The following median (lower/upper quartile) differences across the group were determined: MaxOD -33.8 % (-46.2 to -19.1 %), MeanOD -44.0 % (-54.6 to -26.6 %). Larger changes were observed in elderly patients [<70 years of age (n = 25 eyes): MaxOD -13.4 % (-20.5 to 3.6 %), MeanOD -23.6 % (-30.5 to -15.3 %) versus patients ≥70 years (n = 61 eyes) MaxOD -40.5 % (-53.2 to -30.1 %), MeanOD -47.2 % (-57.8 to -40.1 %)] and in patients with progressed stage of cataract. MaxOD for lens opacification grade 1 (n = 9 eyes): -27.4 % (-42.1 to -19.6 %), grade 2 (n = 26 eyes): -35.0 % (-44.2 to -25.3 %), grade 3 (n = 21 eyes): -34.4 % (-45.4 to -11.4 %), grade 4 (n = 25 eyes): -32.6 % (-53.2 to -6.4 %), and grade 5 (n = 5 eyes): -53.5 % (-61.7 to -38.7 %) and MeanOD for cataract stage 1 (n = 9 eyes): -42.6 % (-46.0 to -26.0 %), stage 2 (n = 26 eyes): -44.1 % (-51.8 to -26.2 %), stage 3 (n = 21 eyes): -45.7 % (-54.7 to -24.7 %), stage 4 (n = 25 eyes): -39.5 % (-59.4 to -26.1 %), and stage 5 (n = 5 eyes): -57.0 % (-66.1 to -51.4 %). CONCLUSIONS: As established by comparison of pre- to post-operative measurements, cataract presented a strong effect on MPOD measured by one-wavelength reflection method. Particular care should therefore be taken when evaluating MPOD using this method in elderly patients with progressed stage of cataract. Future optimization of correcting parameters of scattered light and consideration of cataract influence may allow more precise evaluation of MPOD.

Parametric model for the 3D reconstruction of individual fovea shape from OCT data.

As revealed by optical coherence tomography (OCT), the shape of the fovea may vary greatly among individuals. However, none of the hitherto available mathematical descriptions comprehensively reproduces all individual characteristics such as foveal depth, slope, naso-temporal asymmetry, and others. Here, a novel mathematical approach is presented to obtain a very accurate model of the complete 3D foveal surface of an individual, by utilizing recent developments in OCT. For this purpose, a new formula was developed serving as a simple but very flexible way to represent a given fovea. An extensive description of the used model parameters, as well as, of the complete method of reconstructing a foveal surface from OCT data, is presented. Noteworthy, the formula analytically provides characteristic foveal parameters and thus allows for extensive quantification. The present approach was verified on 432 OCT scans and has proved to be able to capture the whole range of asymmetric foveal shapes with high accuracy (i.e. a mean fit error of 1.40 µm).

Optical coherence tomography as a diagnostic tool for retinal pathologies in avian ophthalmology.

PURPOSE: Optical coherence tomography (OCT) is an established diagnostic tool for retinal pathologies in human eyes and has been adapted to small animal models. However, there have been only a few attempts to use OCT for examination of avian eyes, and little is known about structural details of healthy or pathologically affected retinas in living birds. METHODS: We used SD-OCT (high-resolution spectral domain OCT) to investigate eyes of various avian species including birds of prey. The birds were anesthetized by isoflurane application during OCT examination. Eyes of a common buzzard (Buteo buteo) could be used for a comparative analysis of OCT images and histologic/immunohistochemical examinations. RESULTS: We investigated 45 wild and domestic birds (25 different species, 40 g-7.7 kg body mass) without and with diverse pathologic indications (e.g., body or head trauma). Animals were generally and ophthalmologically examined, and the diagnostic findings of direct ophthalmoscopy and OCT were compared. The OCT examination revealed an increased number of animals with clinical findings and allowed a detailed assessment of structural changes in retinal and choroidal tissue compared to simple direct ophthalmoscopy. Common findings were retinal and choroidal degeneration, retinal detachment, choroidal schisis, drusen, and drusenoid changes. Histologic and immunohistochemical analysis of retinal tissue confirmed the findings of the OCT examination. CONCLUSIONS: Spectral domain OCT of eyes in living birds is applicable and useful as a diagnostic tool in veterinary clinical practices and for vision research in general. Optical coherence tomography improves the quality of the common assessment methods in avian ophthalmology, and expands the diagnostic possibilities with respect to identification and prognosis of diseases. This will be particularly important for hereditary retinal defects, especially of precious breeding individuals, or estimation of treatment success in traumatized wild birds with the aim of release back into the wild.

Optical properties of retinal tissue and the potential of adaptive optics to visualize retinal ganglion cells in vivo.

Many efforts have been made to improve the diagnostic tools used to identify and to estimate the progress of ganglion cell and nerve fibre degeneration in glaucoma. Imaging by optical coherence tomography and measurements of the dimensions of the optic nerve head and the nerve fibre layer in central retinal areas is currently used to estimate the grade of pathological changes. The visualization and quantification of ganglion cells and nerve fibres directly in patients would dramatically improve glaucoma diagnostics. We have investigated the optical properties of cellular structures of retinal tissue in order to establish a means of visualizing and quantifying ganglion cells in the living retina without staining. We have characterized the optical properties of retinal tissue in several species including humans. Nerve fibres, blood vessels, ganglion cells and their cell processes have been visualized at high image resolution by means of the reflection mode of a confocal laser scanning microscope. The potential of adaptive optics in current imaging systems and the possibilities of imaging single ganglion cells non-invasively in patients are discussed.