M. F. Cordeiro

Alzheimer's Disease and Retinal Neurodegeneration

Alzheimers disease (AD) is the major cause of dementia in the world. Although the entorhinal cortex and hippocampal complex are best known as the sites of early pathology in AD, increasing evidence shows that the eye, particularly the retina, is also affected. The AD-related changes in the retina are associated with degeneration and loss of neurons, reduction of the retinal nerve fibres, increase in optic disc cupping, retinal vascular tortusity and thinning, and visual functional impairment. Given the fact that evaluating pathologic changes in the brain during life has always been an indirect process, largely shielded from view by the barrier of the skull, the eye can be used as a window into diseases of the brain. Using modern techniques, the changes in the retina can be visualized in real-time. In addition to the changes in the eyes of AD patients, similar mechanisms of neurodegeneration in the brain have also been demonstrated in the eye. Targeting AD-liked changes in the retina has been recently shown to be effective in the reduction of retinal neuronal degeneration and loss in eye diseases. This review will cover recent findings on retinal degeneration in AD, pathological similarities between AD and eye diseases, and highlight the potential of modern technologies for the detection of prospective biomarkers in the eye in early AD.

Modulation of wound healing after glaucoma surgery.

The healing process after glaucoma filtration is the main determinant of surgical failure and, even more important, the final intraocular pressure. The ability to fully control wound healing may ultimately give us the ability to set the intraocular pressure in the low teens for all patients undergoing glaucoma filtration surgery. The authors review the changes in how to use antimetabolites to improve safety, and many of the exciting new areas of progress, including growth factor neutralization and future molecular therapies to control wound healing.

Imaging multiple phases of neurodegeneration: a novel approach to assessing cell death in vivo.

Nerve cell death is the key event in all neurodegenerative disorders, with apoptosis and necrosis being central to both acute and chronic degenerative processes. However, until now, it has not been possible to study these dynamically and in real time. In this study, we use spectrally distinct, well-recognised fluorescent cell death markers to enable the temporal resolution and quantification of the early and late phases of apoptosis and necrosis of single nerve cells in different disease models. The tracking of single-cell death profiles in the same living eye over hours, days, weeks and months is a significant advancement on currently available techniques. We identified a numerical preponderance of late-phase versus early-phase apoptotic cells in chronic models, reinforcing the commonalities between cellular mechanisms in different disease models. We showed that MK801 effectively inhibited both apoptosis and necrosis, but our findings support the use of our technique to investigate more specific anti-apoptotic and anti-necrotic strategies with well-defined targets, with potentially greater clinical application. The optical properties of the eye provide compelling opportunities for the quantitative monitoring of disease mechanisms and dynamics in experimental neurodegeneration. Our findings also help to directly observe retinal nerve cell death in patients as an adjunct to refining diagnosis, tracking disease status and assessing therapeutic intervention.

Neuroprotection in glaucoma: Drug-based approaches

In recent years the focus of glaucoma research has shifted toward neuroprotection, as the traditional strategies of lowering intraocular pressure have been shown to be unable to prevent progressive vision loss in some glaucoma patients. As a result various neuroprotective drug-based approaches have been shown capable of reducing the death of retinal ganglion cells, which is the hallmark of glaucomatous optic neuropathy. There has been increasing evidence that glaucomatous neurodegeneration is analogous to other neurodegenerative diseases in the central nervous system, with recent work from our group elucidating a strong link between basic cellular processes in glaucoma and Alzheimer’s disease. Additionally, there is a growing trend for using existing neuroprotective strategies in central nervous system diseases for the treatment of glaucoma. In fact, a trial treating patients with primary open-angle glaucoma with memantine, a drug approved for the treatment of Alzheimer’s disease, has recently been completed. Results of this trial are not yet available. In this review, we will examine currently advocated neuroprotective drug-based strategies in the potential management of glaucoma.

Tracking Longitudinal Retinal Changes in Experimental Ocular Hypertension Using the cSLO and Spectral Domain-OCT

PURPOSE Involvement of the outer retina is controversial in glaucoma. The aim of this study was to test, first, whether the outer retina is affected in experimental ocular hypertension (OHT) and, second, whether whole retinal thickness can be used as a surrogate marker of glaucomatous change. METHODS OHT was surgically induced in 20 Dark Agouti rats. Animals were imaged using a modified Spectralis OCT (Heidelberg Engineering, Heidelberg, Germany) at baseline and at 3 and 8 weeks after OHT induction. Measurements were recorded for whole and individual retinal layer thickness in four regions-temporal, superior, nasal, and inferior-around the optic nerve head. RESULTS Whole retinal thickness in normal eyes was 172.19 ± 5.17 μm, with no significant regional differences. OHT caused a significant reduction in whole retinal thickness and the outer nuclear layer (ONL) at 3 and 8 weeks (P < 0.05), along with the expected thinning of the retinal nerve fiber layer (RNFL). Whole retinal thickness correlated well with RNFL (P = 0.035) and ONL (P ≤ 0.001) changes. Sensitivity of RNFL and ONL to IOP exposure appeared greater at 3 than at 8 weeks. In addition, regional profiles were significantly altered in the ONL and RNFL after OHT induction. CONCLUSIONS Adaptation of the Spectralis OCT enables tracking of structural damage in experimental rat OHT. Here the authors show evidence of glaucomatous damage in the outer retinal layers of this model with significant regional changes and highlight whole retinal thickness in the rat as a useful surrogate marker of inner and outer retinal changes. The authors believe that the OCT data can provide useful information with regard to clinical management.

Modulating conjunctival wound healing

Advances in molecular and cell biology have led to an expansion in our knowledge and understanding of the processes involved in wound healing. We review existing and potential therapies modulating the conjunctival scarring response, with particular reference to glaucoma filtration surgery. We discuss how the refinement of present antimetabolite regimens can minimise complications and improve surgical results, and advocate their use in carefully selected patient groups. Perhaps the most promising approach is targeting biological molecules. Hence, use of fully human neutralising monoclonal antibodies to the growth factor TGFfβ has potential as a useful strategy for modifying conjunctival scarring. Combination therapies may also afford an improved therapeutic index. It is hoped that future therapies can offer safer, more specific, focal and titratable treatment, with far-reaching clinical applications.

Imaging apoptosis in the eye

Apoptosis is a form of programmed cell death that is implicated in both pathological and physiological processes throughout the body. Its imaging in vivo with intravenous radiolabelled-annexin V has been heralded as an important advance, with around 30 clinical trials demonstrating its application in the early detection and monitoring of disease, and the assessment of efficacy of potential and existing therapies. A recent development has been the use of fluorescently labeled annexin V to visualize single retinal cells undergoing the process of apoptosis in vivo with ophthalmoscopy. This has been given the acronym DARC (Detection of Apoptosing Retinal Cells). DARC so far has only been used experimentally, but clinical trials are starting shortly in glaucoma patients. Results suggest that DARC may provide a direct assessment of retinal ganglion cell health. By enabling early assessment and quantitative analysis of cellular degeneration in glaucoma, it is hoped that DARC can identify patients before the onset of irreversible vision loss. Furthermore, in addition to aiding the tracking of disease, it may provide a rapid and objective assessment of potential and effective therapies, providing a new and meaningful clinical endpoint in glaucomatous disease that is so badly needed.

Localisation and significance of in vivo near-infrared autofluorescent signal in retinal imaging

Aim To evaluate near-infrared (NIR) autofluorescence (AF) in patients with geographic atrophy (GA) secondary to age-related macular degeneration and to investigate the origin of the signal by in vivo and histological analysis in rats and in a human donor eye. Methods Confocal scanning laser ophthalmoscopy in vivo imaging, including blue (excitation: 488 nm, emission 500–700 nm) and NIR (excitation: 790 nm, emission >810 nm) AF was performed in 21 eyes of 18 GA patients. Pigmented and albino rats underwent with the same device both in vivo and post-mortem imaging. For the latter, cryostat prepared retinal cross-sections were imaged using an additional customised magnification lens. Finally, cross-sections of a 49-year old human donor eye were recorded. Results Atrophic areas in GA were characterised by low NIR AF intensities. In the junctional zone of atrophy, focal areas of increased intensity were seen which appeared to seldom correlate to blue AF findings. Confocal live scanning in pigmented rats identified the maximum of the NIR AF signal in the outer retina, with histological confirmation of the signal origin localised to the retinal pigment epithelium and sclera in both animals and human donor eye. No NIR AF was found in the retina of young non-pigmented rats. Discussion This study further underscores the assumption that melanin is the main source of NIR AF in the healthy retina. Increased NIR AF intensities in the junctional zone in GA may represent accumulation of melanolipofuscin, which may reflect disease activity and thus may allow for early identification of patients at high-risk of GA enlargement.

Molecular therapy in ocular wound healing

Recent advances in molecular cell biology have revolutionised our understanding of medical diseases and provided new and alternative strategies for developing treatments. Here we review the spectrum of molecular therapies that are either currently available or have potential application as agents that are able to modulate the wound healing response in the eye. For the purposes of this review, we define molecular therapy as the targeting of specific molecules known to be involved in the processes of wound healing. This may be at the level of either protein or gene expression. The process of wound healing is involved in either the pathogenesis or failure of treatment of many of the major blinding or visually disabling conditions in the world today. It is implicated in scarring diseases throughout the eye, some examples of which are described below. The conjunctival wound healing response is important in many ocular conditions such as pemphigoid, trachoma, and chronic cicatrisation, where the development of complications arises from the disruption of the ocular surface.1 It is also a major determinant of outcome following glaucoma filtration and squint surgery.2The severity and extent of clinical disease are closely related to the degree of conjunctival scarring. Another example of scarring is that occurring in the cornea after excimer laser photorefractive keratectomy (PRK), giving rise to symptoms of haze and resulting, in some cases, in a reduction of the best corrected visual acuity.3 4 The scarring condition of proliferative vitreoretinopathy (PVR) accounts for 7–10% of surgical failures in primary retinal detachment repair procedures5 6 and is responsible for producing significant visual morbidity and blindness. It is characterised by the development of fibrosing and proliferating cellular membranes on the vitreous and retinal surfaces that contract and cause irreparable tractional retinal detachments. The complex process of wound healing …

En face optical coherence tomography: a new method to analyse structural changes of the optic nerve head in rat glaucoma.

Aim: To investigate en face optical coherence tomography (eOCT) and its use as an effective objective technique for assessing changes in the glaucomatous rat optic nerve head (ONH) in vivo, and compare it with confocal scanning laser ophthalmoscopy (cSLO). Methods: 18 Dark Agouti (DA) rats with surgically induced ocular hypertension were imaged with eOCT and cSLO at regular intervals. Assessment included three dimensional (3D) topographic reconstructions, intensity z-profile plots, a new method of depth analysis to define a “multilayered” structure, and scleral canal measurements, in relation to the degree of intraocular pressure (IOP) exposure. Results: The increased depth resolution of the eOCT compared to the cSLO was apparent in all methods of analysis, with better discrimination of tissue planes. This was validated histologically. eOCT demonstrated several significant changes in imaged rat ONH which correlated with IOP exposure, including the area of ONH (p<0.01), separation between retinal vessel and scleral layers (p<0.05), and anterior scleral canal opening expansion (p<0.05). Conclusion: eOCT appears to be effective in assessing rat ONH, allowing detailed structural analysis of the multilayered ONH structure. As far as the authors are aware, this is the first report of scleral canal expansion in a rat model. They suggest eOCT as a novel method for the detection of early changes in the ONH in glaucoma.