Andreas W. A. Weinberger

Classification of abnormal fundus autofluorescence patterns in the junctional zone of geographic atrophy in patients with age related macular degeneration

Aim: To describe and classify patterns of abnormal fundus autofluorescence (FAF) in the junctional zone of geographic atrophy (GA) in patients with age related macular degeneration. Methods: Digital FAF images were recorded in 164 eyes of 107 patients using a confocal scanning laser ophthalmoscope (cSLO; excitation 488 nm, detection above 500 nm) as part of a prospective multicentre natural history study (FAM Study). FAF images were obtained in accordance with a standardised protocol for digital image acquisition and generation of mean images after automated alignment. Results: Image quality was sufficient for classification of FAF patterns in 149 eyes (90.9%) with lens opacities being the most common reason for insufficient image quality. Abnormal FAF outside GA in 149 eyes was classified into four patterns: focal (12.1%), banded (12.8%), patchy (2.0%), and diffuse (57.0%), whereby 12.1% had normal background FAF in the junctional zone. In 4% there was no predominant pattern. The diffuse pattern was subdivided into four groups including reticular (4.7%), branching (27.5%), fine granular (18.1%), and fine granular with peripheral punctate spots (6.7%). Conclusions: Different phenotypic patterns of abnormal FAF in the junctional zone of GA can be identified with cSLO FAF imaging. These distinct patterns may reflect heterogeneity at a cellular and molecular level in contrast with a non-specific ageing process. A refined phenotypic classification may be helpful to identify prognostic determinants for the spread of atrophy and visual loss, for identification of genetic risk factors as well as for the design of future interventional trials.

Macular translocation for surgical management of subfoveal choroidal neovascularizations in patients with AMD: first results

Abstract · Background: At present no satisfying treatment for subfoveal choroidal neovascularization (CNV) secondary to age-related macular degeneration (AMD) is available. Visual results after successful surgical removal of subfoveal CNV are disappointing. This has been explained by a primary dysfunction of the retinal pigment epithelium (RPE) in the macular region and the surgical trauma to the RPE in patients with AMD. Therefore, Machemer and Steinhorst developed a technique for macular translocation after surgical removal of subfoveal CNV. We report our first experiences with this technique in patients with subfoveal CNV secondary to AMD. · Methods: Seven patients aged between 71 and 83 years with subfoveal CNV were included in the study. Visual acuity of the fellow eyes was below 20/400. All patients underwent pars plana vitrectomy. Retinal detachment was produced by subretinal infusion of balanced salt solution and a 360° retinotomy at the base of the vitreous was performed. After removal of the CNV, retinal rotation and reattachment, the retina bordering the retinotomy was coagulated with endolaser photocoagulation. Silicone oil was used as temporary tamponade. · Results: In all patients the subfoveal CNV was removed and the macula was translocated by a 15°–45° rotation onto functional RPE. The mean duration of follow-up was 11±3 months. Initial visual acuity ranged from 20/80 to hand movements. Final visual acuity was 20/100 to 20/400. Initially all patients complained of tilted vision. During follow-up the rotation of the image regressed and was well tolerated by all patients. Complications included the development of retinal detachment in three patients after silicone oil removal, development of a macula pucker, and a significant increase of lens opacity in the phakic eyes. · Conclusion: In our series rapid improvement of visual function was observed in one patient only, even if the macula appeared ophthalmoscopically and angiographically normal. Vitreoretinal complications occurred frequently during follow-up.

CFH, C3 and ARMS2 are significant risk loci for susceptibility but not for disease progression of geographic atrophy due to AMD

Background Age-related macular degeneration (AMD) is a prevalent cause of blindness in Western societies. Variants in the genes encoding complement factor H (CFH), complement component 3 (C3) and age-related maculopathy susceptibility 2 (ARMS2) have repeatedly been shown to confer significant risks for AMD; however, their role in disease progression and thus their potential relevance for interventional therapeutic approaches remains unknown. Methodology/Principal Findings Here, we analyzed association between variants in CFH, C3 and ARMS2 and disease progression of geographic atrophy (GA) due to AMD. A quantitative phenotype of disease progression was computed based on longitudinal observations by fundus autofluorescence imaging. In a subset of 99 cases with pure bilateral GA, variants in CFH (Y402H), C3 (R102G), and ARMS2 (A69S) are associated with disease (P = 1.6×10−9, 3.2×10−3, and P = 2.6×10−12, respectively) when compared to 612 unrelated healthy control individuals. In cases, median progression rate of GA over a mean follow-up period of 3.0 years was 1.61 mm2/year with high concordance between fellow eyes. No association between the progression rate and any of the genetic risk variants at the three loci was observed (P>0.13). Conclusions/Significance This study confirms that variants at CFH, C3, and ARMS2 confer significant risks for GA due to AMD. In contrast, our data indicate no association of these variants with disease progression which may have important implications for future treatment strategies. Other, as yet unknown susceptibilities may influence disease progression.

Iris pigment epithelial cell translocation in exudative age-related macular degeneration. A pilot study in patients.

Abstract Background: This prospective, non-controlled pilot study investigates the practicability of IPE translocation and functional outcome in ARMD patients. Removal of submacular choroidal neovascularization (CNV) in age-related macular degeneration (ARMD) is usually associated with RPE damage and poor visual prognosis. Homologous RPE transplants fail to preserve macular function, possibly due to immune rejection. Instead of homologous RPE, we suggest translocating autologous iris pigment epithelium (IPE), building on earlier evidence from animal and in vitro investigations that IPE can substitute RPE functions in the experimental animal. Immunological cell rejection is avoided. Methods: Four eyes with well- defined and eight eyes with ill- defined subfoveal CNV were submitted to operation and followed up for a minimum of 6 months. IPE cells were harvested from a peripheral iridectomy. A vitrectomy was performed. Submacular membranes were removed, and isolated IPE cells were injected into the subretinal space. Examinations included ETDRS visual acuity, fluorescein angiography, and SLO microperimetry. Results: All patients underwent successful surgical removal of CNV and subretinal IPE injection. Compared to preoperative visual acuity (20/400–20/100) no significant change was observed after 6 months (20/320–16/80). A change of more than two ETDRS chart lines was defined as significant. One eye with preoperative ill-defined CNV developed a recurrence, leading to reduced visual acuity. In all patients, postoperative fluorescence angiography revealed early hyperfluorescence (window defect) in the surgically denuded area. Central fixation was demonstrated in 50% of eyes. Conclusions: Preliminary data suggests that IPE translocation in submacular surgery for ARMD can preserve but not improve preoperative visual acuity over 6 months. Functional results are promising compared to submacular membrane extraction alone and RPE transplantation. Continued research on improvement of IPE translocation seems justified.

Translocation of iris pigment epithelium in patients with exudative age-related macular degeneration: long-term results

PurposeTo report the practicability and efficacy of autologous iris pigment epithelium (IPE) translocation in exudative age-related macular degeneration (ARMD) over 1 year.MethodsThe consecutive interventional case series included 56 patients with exudative ARMD. During vitrectomy the submacular neovascular membrane (CNV) was removed and IPE cells, harvested from a peripheral iridectomy, were injected into the submacular space. Included were patients with subfoveal occult CNV (11 eyes), classic CNV (10 eyes), mixed CNV (17 eyes), CNV with a pigment epithelial detachment (13 eyes) or CNV with a hemorrhage (5 eyes). Outcome measures were visual acuity, foveal fixation, size of CNV and rate of recurrence based on fluorescence angiographic imaging.ResultsAll patients underwent successful surgical removal of the CNV with consecutive subretinal IPE injection. Visual acuity was better than 20/100 in 19 patients preoperatively and in 18 patients postoperatively. A visual acuity of 20/100 or less was found in 37 patients preoperatively and in 38 patients postoperatively. Mean preoperative visual acuity (1.0±0.3 logMAR units) did not change significantly after 1 year (1.0±0.3 logMAR units). Ten eyes (18%) developed a recurrence. Fixation within the surgically denuded area could be demonstrated in 25 eyes (45%).ConclusionsAutologous IPE translocation for ARMD over one year can preserve foveal function on a low level, but cannot improve visual acuity. IPE translocation is technically feasible with a low rate of complications. Continued research seems justified to improve functional outcome.

Effects of Bevacizumab (Avastin) on Retinal Cells in Organotypic Culture

PURPOSE Repetitive intravitreal injections of bevacizumab are a successful treatment option for exudative age-related macular degeneration (AMD). The aim of this study was to evaluate the toxicity of bevacizumab in the adult mammalian neurosensory retina in culture. METHODS Adult porcine neurosensory retinas were cultured adjoined to the retinal pigment epithelium-choroid layer (retina-RPE-choroid complex) in static culture for 3 days, whereas neural retinas alone were cultured in a perfusion chamber for 3 days. Bevacizumab was added to the culture and perfusion medium at three concentrations (0.25 mg/mL [n = 6], 0.5 mg/mL [n = 6], and 1.25 mg/mL [n = 6]). Retina-RPE-choroid complex and neural retinas alone cultured without bevacizumab were used as controls. After 3 days in culture, the neural retinas alone and the retina-RPE-choroid complexes were analyzed histologically and immunohistochemically for the expression of glial fibrillary acidic protein (GFAP), vimentin, glutamine synthetase, rhodopsin, smooth muscle actin (SMA), and apoptosis. RESULTS No toxic effects on ganglion or photoreceptor cells were observed at any concentration of bevacizumab. The expression of GFAP and vimentin was slightly increased in Müller cells, whereas glutamine synthetase and rhodopsin were unaffected by bevacizumab. However, significantly enhanced SMA expression in retina blood vessels was observed in retinas cultured in the presence of bevacizumab. CONCLUSIONS Bevacizumab was well tolerated by ganglion and photoreceptor cells even at concentrations fivefold higher than those used clinically. The increased expression of SMA is an indication of the loss of functional VEGF modulating smooth muscle cells in mature vessels.

Effects of Fibrinogen and α2-Macroglobulin and Their Apheretic Elimination on General Blood Rheology and Rheological Characteristics of Red Blood Cell Aggregates

Methods of therapeutic apheresis, such as plasma exchange or rheopheresis eliminate moderately aggregating macromolecules like fibrinogen, as well as strongly aggregating substances like α2‐macroglobulin from blood. In order to examine the specific effect of eliminating α2‐macroglobulin as a highly aggregating macromolecule, this study aimed to analyze the different rheological properties of: (i) moderately aggregating red blood cells (RBCs; inducible by fibrinogen); and (ii) strongly aggregating RBCs (inducible by α2‐macroglobulin). In vitro, RBC aggregate geometry was determined in the presence of strong and moderate aggregation inducing macromolecules. In vivo, flow behavior of RBC aggregates was analyzed by intravital microscopy. Using network scanning, the number of perfused and non‐perfused microvessels was determined. In vitro, the higher adhesive forces of strongly aggregating RBCs led to both a higher packing density of single RBCs within aggregates, expressed as a significantly reduced thickness of individual RBCs, and greater deformation, expressed as a significantly diminished offset between RBCs and an increased curvature of RBCs at the ends of the aggregates. In vivo rheoscopy showed that only high aggregating RBCs persisted in the precapillary bed and led to the absence of RBCs in up to 40% of nutritive capillaries. These novel findings are of importance regarding recent developments in clinical hemorheology, specifically the clinical use of hemapheretic therapies for diseases in which impaired microcirculation plays a role in either their development or progression, such as age‐related macular degeneration and complications of diabetes mellitus. Our data support that procedures reducing the concentration of α2‐macroglobulin in blood by extracorporeal elimination might provide a more efficient improvement of overall blood fluidity in microcirculatory beds.

Improving the quality of multifocal visual evoked potential results by calculating multiple virtual channels.

PurposeTo introduce a method for improvement of multifocal VEP (mfVEP) recordings by prediction of waveforms at multiple positions on the surface of the skull.MethodsFifteen healthy participants (mean age 24 ± 3.8 years) underwent mfVEP recordings from 3 surface positions. Two methods of a best-of-mfVEP approach were used and compared. In the first, a standard procedure, further data from 3 calculated channels were used. In the second approach, mfVEPs were obtained by using data derived from 40 virtual electrode positions on the basis of predictions from dipole source calculations.ResultsThe mean signal-to-noise ratios (SNRs) of the best-of-mfVEPs of both methods were compared. The SNR was significantly higher for mfVEP data using additional virtual recordings revealed by dipole source determination (2.87 vs. 3.36; P < 0.035).ConclusionWe conclude that multichannel prediction of mfVEP responses based on dipole source calculation significantly improves the quality of the examination results compared with the currently prevalent standard method.

Surface-modified silicone foils for intraocular implantation

BackgroundImplantation of silicone materials like iris diaphragms into the eye can be complicated by cell migration and attachment. We studied polydimethylsiloxane (PDMS) foils coated with isocyanate terminated, star-shaped poly(ethylene glycol-stat-propylene glycol) (NCO-sP(EO-stat-PO)) equipped with heparin towards the inhibition of cell attachment without influencing cell viability.MethodsMouse fibroblasts L929 were cultured and seeded onto sterilized pieces of either uncoated NCO-sP(EO-stat-PO) or heparin-NCO-sP(EO-stat-PO) loaded foils. Polyvinylchloride (PVC) foils served as the positive control and biomembranes as the negative control. The cultured cells were examined after 24 h for cell viability and adhesion by fluorescence microscopy; morphological cell changes were documented after hemalaun staining. Cell density was measured and quantification of cell proliferation was assessed by a BrdU test; quantification of cell activity was analyzed by a WST-1 test.ResultsThe fibroblasts’ cell viability was excellent on all tested foils except the toxic PVC foil. NCO-sP(EO-stat-PO) coating provided significantly reduced cell activity. On heparin-loaded coatings, cells were viable and less dense but showed almost the same cell proliferation and cell activity as on the negative control. NCO-sP(EO-stat-PO) coated, heparin loaded foils proved high biocompatibility and reduced cell adhesion.ConclusionsBoth NCO-sP(EO-stat-PO)-coated foils with and without heparin seemed to be a viable implantation material for less cell migration, attachment, and reduced implant complications. Conclusive we give a recommendation for further studies on the intraocular implantation in particular for the NCO-sP(EO-stat-PO)-coated foils.

Fluoreszenzangiographie bei altersabhängiger Makuladegeneration

ZusammenfassungDa weder eine kurative Therapie noch eine sichere Prophylaxe bekannt ist, wird gegenwärtig versucht, das Fortschreiten der Erkrankung aufzuhalten. Ein frühzeitiger Therapiebeginn ist daher angezeigt. In diesem Zusammenhang kommt der Klassifikation der verschiedenen Formen der altersabhängigen Makuladegeneration eine wichtige Rolle zu. In den folgenden Kapiteln soll anhand von klinischen Beispielen dargestellt werden, wie fluoreszenzangiographische Befunde bei AMD klassifiziert und interpretiert werden.