[Are There Different Phenotypes in Geographic Atrophy of AMD? - Pilot Study on Differentiation Using Multimodal Imaging]. / Gibt es unterschiedliche Phänotypen bei der geografischen Atrophie der AMD? ­ Pilotstudie zur Differenzierung mittels multimodaler Bildgebung.

BACKGROUND: Geographic atrophy (GA) in patients with age-related macular degeneration (AMD) involves a loss of photoreceptors (PR), retinal pigment epithelium (RPE) and choriocapillaris (CC). For treatment decisions, it is crucial to discern which of these layers the damage originates, subsequently spreading to the others. It has long been thought that the RPE, with its accumulation of lipofuscin, is the site of primary damage in the development of GA. However, histological studies have shown that in some patients, the PR are affected first, followed by secondary damage to the RPE and CC, and in others regression of the CC is the first manifestation. The aim of this study was to use multimodal imaging to determine the extent of the damage at the levels of the PR, RPE and CC, to characterise the individual phenotypic variations of GA and to investigate the corresponding functional impairment. PATIENTS AND METHODS: Twenty eyes of 20 patients (mean age 78 years; 14 female, 6 male) with the clinical diagnosis of GA were examined by means of fundus autofluorescence (FAF) to evaluate the damage to the RPE, en face SD-OCT at the level of the PR to characterise the area of cell loss in this layer and OCT angiography (OCT-A, AngioVue, Optovue; 50 µm CC-segmentation with localization below the RPE) to assess regression of the CC. The affected area of each layer was measured. Best-corrected visual acuity (BCVA) test and fundus correlated automated 10° microperimetry (MAIA Microperimetry, CENTERVUE; 4-2 strategy, 68 stimuli) were performed in all patients. The results of these examinations were evaluated and correlated. RESULTS: All eyes showed a different extent of the areas of atrophy in the PR, RPE and CC. The layer with the largest area of atrophy was the RPE in 13 eyes (65%), the PR in 3 eyes (15%) and the CC in 4 eyes (20%). While the visual loss depended entirely on the presence of foveal sparing, microperimetry revealed a correlation between the extent of detectable functional deficit and the largest atrophic area. CONCLUSIONS: Multimodal imaging with FAF, en face OCT, OCT-A and a correlation with microperimetry enables a clinical phenotypic differentiation in GA as well as a more precise characterisation of the associated functional impairment. This confirms clinically the histologically demonstrated diversity of the damaged structure (PR, RPE or CC) in patients with GA. However, the variations identified in this pilot study must be confirmed in Reading Center-based larger cohorts. The approach described here may lead to differentiated consideration of the anatomical and functional aspects of the disease and turn out to be helpful in patient selection as well as in identifying and monitoring future therapeutic approaches.

[Analysis of the "Portal" Care Model - Examination of the Outcome Quality of IVOM Therapy with Regard to Latency Periods in Exudative AMD]. / Analyse des Versorgungsmodells "Portal" ­ Untersuchung der Ergebnisqualität der IVOM-Therapie im Hinblick auf Latenzzeiten bei exsudativer AMD.

BACKGROUND: For many maculopathies, the management of intravitreal injection (IVOM) presents a logistical challenge. To ensure contemporary and timely treatment, patients have to organise their rides to the surgery, and the clinic has to provide enough short term resources. The objective of this study is an evaluation of the IVOM therapy for patients with exudative AMD according to four quality indicators a) latency time within the treatment and monitoring cycle, b) the treatment and monitoring frequency, c) the adherence and d) the medical outcome. MATERIALS AND METHODS: For more than seven years, patients with exudative AMD have been treated by many ophthalmologists using a networked portal system. Therefore, conservative doctors and surgical eye centres exchange treatment-relevant data. In total there are documented 2283 eyes of 1850 patients. We evaluate these electronic medical records retrospectively according to the mentioned quality indicators. RESULTS: This evaluation results in a latency time from OCT monitoring and the start of a new IVOM series of 8.1 working days. Within the first two treatment years, we achieve 10.5 injections and 8.2 monitoring visits in average. After two years, 72.9% of the cases were still in treatment or monitoring. We observed stabilisation of mean visual accuracy of about 0.05 logMAR. CONCLUSIONS: To improve the visual acuity, it is essential to achieve consistent therapy over a long period of time, especially in the case of treatment-relevant exudative AMD. The evaluation of our treatment system demonstrated that the PRN-scheme can be implemented by a cooperatively organised IVOM therapy. It is possible to achieve rapid retreatment and good adherence over many treatment years. For treatment-relevant exudative AMD it is essential for the improvement of the visual accuracy to implement consistent therapy over a long period of time. The evaluation of our treatment system demonstrates that the PRN scheme can be implemented in a cooperatively organised IVOM therapy. It is possible to achieve rapid retreatment and good patients' adherence over many treatment years.

NEGATIVE EFFECTS OF VITAL DYES AFTER UNEVENTFUL VITREOMACULAR SURGERY.

PURPOSE: Vital dyes have become a clinical standard during chromovitrectomy but toxicity remains an issue. We compared the clinical outcome of one supposedly toxic vital dye (AV 17 with 5% mannitol) with a standard vital dye (MBB Dual) and performed a power analysis for future comparative studies. METHODS: Retrospective analysis of 270 eyes after chromovitrectomy with internal limiting membrane peeling because of macular holes. Primary endpoint was loss in BCVA >2 lines and photoreceptor atrophy as seen on optical coherence tomography examination. RESULTS: In 173 eyes, staining of the epiretinal membrane and extracellular matrix was performed using MBB (Group A), and in 97 using AV 17-M (Group B). The mean BCVA was not significantly different after more than 3 months and also not in the early postoperative period after surgery between Group A and Group B. The number of patients suffering from a decline in BCVA of two lines and more was not significantly higher in patients of Group B. There was not a significantly higher percentage of patients with an inner segment/outer segment defect. CONCLUSION: Our rather homogeneous study showed no significant difference between both dyes. Thousand five hundred patients would need to be examined to find a significant difference in future studies.

Characteristics of type 1 and 2 CNV in exudative AMD in OCT-Angiography.

BACKGROUND: With FA and SD-OCT, different types of CNV in exudative AMD may be differentiated: type 1 CNV (within the sub-RPE space, typically corresponding to angiographically occult CNV), type 2 CNV (within the subretinal space, typically corresponding to angiographically classic CNV) and type 3 NV (intraretinal retinal angiomatous proliferation). OCT-angiography (OCT-A) is a new method to visualize vasculature based on flow characteristics. A correlation of type 1 and 2 CNV was performed. METHODS: Thirty-six eyes (17 type 1 CNV, 9 combined type 1 and 2 CNV, and 10 type 2 CNV) of 36 patients were examined by FA, SD-OCT and OCT-A. Standardized OCT-A segmentations were performed at the level of mid-choroid, choriocapillaris (CC), RPE and outer retina. On these images the size and demarcation of CNV lesions were classified: "not distinguishable", "minor" or "sharp" demarcation. Furthermore, the size of the different CNV subtypes was determined and compared. RESULTS: Both types of CNV were visible in OCT-A. They could be detected on the slabs "mid-choroid", "CC" and "RPE". While type 1 CNV showed most often a minor demarcation from the surrounding vasculature, type 2 CNV showed nearly always a sharp demarcation. In addition, type 2 CNV extended into the slab "outer retina" and were much smaller than type 1 CNV. CONCLUSIONS: Different types of CNV in exudative AMD can be visualized and differentiated with OCT-A. Type 1 CNV were larger with minor demarcation from the surrounding vasculature and were visible on the slab "mid-choroid", "CC" and "RPE". In contrast, type 2 CNV demonstrated a sharp demarcation from the surrounding vasculature reaching the slab "outer retina".

[Exudative AMD in OCT Angiography]. / Exsudative AMD in der OCT-Angiografie.

OCT angiography provides a combination of vascular and structural information. In contrast to conventional fluorescein angiography, there is no need for dye injection to give an image of choroidal neovascularisation. Although there is currently no classification of OCT angiography, different neovascular patterns can be observed, with criteria for activity. In the future, OCT angiography may help us to understand pathophysiological mechanisms and to develop therapeutic strategies.

[Changes in OCT Angiography of Type 2 CNV in Neovascular AMD during Anti-VEGF Treatment]. / OCT-angiografische Veränderungen von der Typ-2-CNV bei exsudativer AMD unter Anti-VEGF-Therapie.

Purpose OCT-A is a new method to visualise the 2D and 3D structures of neovascular complexes in exudative AMD. The aim of the present study was to characterise type 2 CNV in different 2D segmentations and in 3D imaging and to investigate changes during anti-VEGF treatment. Methods 12 patients with type 2 CNV in FA and SD-OCT were selected. OCT-A (Avanti, Optovue) was obtained initially and after the first three injections and thereafter, if "new activity" (increase in sub- or intraretinal fluid) occurred. The characteristics of the type 2 CNV were classified initially and during follow-up in different segmentations (outer retina, RPE, CC, choroidea), in respect to the size of the CNV, the flow area within the CNV and flow index (% of flow area within the total lesion). Results Comparison of the vessel characteristics before and after anti-VEGF treatment showed a significant reduction in the size of CNV at every level (p < 0.05). This was most significant at the RPE level (p < 0.005). After new activity, a significant increase in size was only recognised at the CC level (p < 0.05). Similarly, the most significant changes in the flow area were measured at the RPE level before and after treatment (p < 0.01) and at the CC level after new activity (p < 0.05). Demarcation from type 2 CNV of the bordering tissue was much better when activity occurred. Conclusions OCT-A provides a new opportunity for the assessment of vascular characteristics of type 2 CNV, and quantifies CNV size and vascularisation under anti-VEGF therapy. This may be used in further studies in combination with SD-OCT scans to describe characteristics of this type of CNV under treatment. OCT-A is an additional medical imaging procedure to SD-OCT and FA, but more experience is needed in distinguishing CNV in the active and non-active stages.

[OCT Angiography in Exudative AMD with Detachment of Vascularised Retinal Pigment Epithelium]. / OCT-Angiografie bei exsudativer AMD mit vaskularisierter Pigmentepithelabhebung.

Background The aim of the following extended case study was to analyse whether choroidal neovascularisation (CNV) in vascularised epithelial detachments (PED) in OCT angiography (OCT-A) can be better visualised in OCT-A than in the established angiographic methods during the course of anti-VEGF therapy and if possible used to quantify the CNV size and flow area. These findings were compared with other SD-OCT characteristics of the lesion (PED height, retinal thickness). Patients and Methods 8 patients with PED and associated CNV were diagnosed with multimodal imaging and additionally OCT angiography was performed. The CNV region in the B-scan of the OCT-A was detected with a fine segmentation setting (20 µm) parallel and just below the retinal pigment epithelium (RPE). The CNV area was manually marked, and the size of the CNV and the vessel section (flow area) were analysed with the evaluation tool of the device. This measurement was performed both initially and after anti-VEGF therapy (3 injections). At the same time, the visual acuity (logMAR) and the SD-OCT parameters of PED height and retinal thickness were determined before and after therapy and also statistically compared. Results Initially, the size of CNV in OCT-A showed a large phenotypic range of variation (0.33 - 1.35 mm2, mean 0.71 mm2). This decreased significantly under therapy (after therapy 0.44 - 0.84 mm2, mean 0.57 mm2, p = 0.02). The proportion of the vessels analysed within the CNV (flow area) varied as well (0.21 - 0.88 mm2, mean 0.45) and decreased under therapy (0.08 - 0.44 mm2 after therapy), mean 0.27 mm2, p = 0.07). The height of PED in SD-OCT was initially different (initially 274 - 1459 µm, mean 607 µm), but showed only small changes (132 - 1317 µm, mean 524 µm, p = 0.09) under therapy. This also applied to the mean retinal thickness (before therapy 315 µm, after therapy 294 µm, p = 0.5). Mean visual acuity also improved only slightly (p = 0.7) after therapy. from initially 0.51 to 0.45 logMAR. Conclusions The combination of SD-OCT and OCT-A offers significantly improved visualisation and quantification of CNV in a vascularised PED. With the help of OCT-A imaging, changes in the perfusion/size of the CNV can be quantified. Together with the retinal activity signs, this allows a second activity assessment of the CNV under anti-VEGF therapy. Due to its three-dimensional structure, especially for this subtype of the exudative AMD, it is of the utmost importance to develop three-dimensional imaging for both structural SD-OCT and the OCT-A.

[OCT Angiography of RPE Tears in Exudative AMD: Morpohological Analysis of the Choriocapillaris and the RPE]. / OCT-Angiografie bei RPE-Einrissen im Rahmen einer exsudativen AMD: morphologische Analysen der Choriokapillaris und des RPE.

Background Retinal pigment epithelium (RPE) tears are a typical complication of vascular pigment epithelium detachment in age-related macular degeneration (AMD). During proactive intense anti-VEGF therapy, stabilisation or improvement of function may occur. With the new method of OCT angiography (OCT-A), retinal vessels and flow density can be quantified. This pilot study investigates changes in the choriocapillars (CC) in areas with increasing FAF in OCT following an RPE-tear. Methods In six eyes with an RPE-tear, prospectively initially and every three months thereafter, multimodal imaging was performed, including fundus autofluorescence (FAF) (HRA2, Heidelberg Engineering, Heidelberg, Deutschland) and OCT-A (RTVue XR Avanti, SSDA-Modus, Angiovue, Optovue, Freemont, CA, USA). With interactive MATLAB-software (MATLAB, MathWorks, Natick, MA, USA), FAF and OCT were geometrically superimposed. With the help of the Fiji software (National Institutes of Health, Bethesda, MD, USA), areas with increasing FAF flow intensity in OCT-A with CC-segmentation were measured during an average follow-up period of 12 months. Results We measured a reduction in the RPE-free area - due to an increase in autofluorescence tissue - of an average of 2.94 mm2 (SD 2.1 mm2; 42.1% of initial RPE-free area) in the boundary area of RPE-tears. At the end of the different follow-ups, some patients exhibited lower flow density in areas of regenerated autofluorescence than the initial findings. On the other hand, in some follow-ups, the same or increased flow density was seen. Conclusion In this pilot study, OCT-A was tested to analyse the structure of CC in areas of regenerated FAF after RPE-tears. Using external image editing software, FAF and OCT-A were compared during the follow-up. Thus apparent initial regression of the CC in the area mentioned above could be observed. During the follow-up and development of autofluorescent SHT, CC also regenerates up to the level of the initial findings of CC.

RETINAL PIGMENT EPITHELIAL TEAR AND ANTI-VASCULAR ENDOTHELIAL GROWTH FACTOR THERAPY IN EXUDATIVE AGE-RELATED MACULAR DEGENERATION: Clinical Course and Long-Term Prognosis.

BACKGROUND: To document the long-term outcome in cases of retinal pigment epithelial (RPE) tears after treatment of vascularized pigment epithelial detachments with anti-vascular endothelial growth factor therapy. METHODS: A retrospective analysis of the long-term outcome of a consecutive series of eyes with RPE tear developed during anti-vascular endothelial growth factor therapy for pigment epithelial detachment associated with choroidal neovascularization or retinal angiomatous proliferation (vascularized pigment epithelial detachment) was performed. Best-corrected visual acuity (BCVA), spectral domain optical coherence tomography, and autofluorescence images and also fluorescein angiograms were analyzed to determine the functional and morphologic development over time. RESULTS: The long-term outcome of 22 eyes (21 patients, 13 women and 8 men; 65-85 years; mean: 76 years) with RPE tear was performed with minimal follow-up of 3 years (range: 3-5 years, mean: 44 months) and re-treatment with different therapeutic strategies. The eyes were differentiated in 2 groups according to the course of BCVA after the first 2 years of follow-up: Group 1 (11 eyes) demonstrated a stabilized or improved BCVA after 2 years and Group 2 (11 eyes) demonstrated a decrease in BCVA after 2 years. The initial BCVA between both groups was comparable. Also the mean initial size of the RPE tear was the same between the 2 groups, the area of the RPE tear decreased continuously during follow-up in Group 1, whereas this was the case in Group 2 only at the beginning of treatment with a further increase of the size of the RPE tear with longer follow-up. This corresponded with a different morphologic development between the two groups. In Group 1, increasing recovery of autofluorescence at the RPE-free area was visible beginning from the outer border, whereas in Group 2, further growth of the neovascular complex in the area of the RPE tear was observed resulting in larger fibrovascular scars. In addition, in both groups, the development of hyperreflective tissue was seen on spectral domain optical coherence tomography in the RPE-free area. The major therapeutic difference between the 2 groups was a significantly larger number of injections especially during the first year in Group 1. CONCLUSION: The development of RPE tear after anti-vascular endothelial growth factor therapy for vascularized pigment epithelial detachment in exudative age-related macular degeneration does not necessarily result in large disciform scars and functional loss, but multiple injections seem to be beneficial especially in the first year. With this strategy, RPE tears seem to be covered by autofluorescent and hyperreflective tissue and a regrowth of the neovascular complex can be prohibited. As a result, photoreceptor cells regain their metabolic support with functional recovery.

Relationship Between Nerve Fiber Layer Hemorrhages and Outcomes in Central Retinal Vein Occlusion.

Purpose: To evaluate the depth and pattern of retinal hemorrhage in acute central retinal vein occlusion (CRVO) and to correlate these with visual and anatomic outcomes. Methods: Retinal hemorrhages were evaluated with color fundus photography and fluorescein angiography at baseline and follow-up. Snellen visual acuity (VA), central foveal thickness (CFT), extent of retinal ischemia, and development of neovascularization were analyzed. Results: 108 eyes from 108 patients were evaluated. Mean age was 63.6 ± 16.1 years with a predilection for the right eye (73.1%). Average follow-up was 17.2 ± 19.2 months. Mean VA at baseline was 20/126 and 20/80 at final follow-up. Baseline (P = 0.005) and final VA (P = 0.02) in eyes with perivascular nerve fiber layer (NFL) hemorrhages were significantly worse than in eyes with deep hemorrhages alone. Baseline CFT was greater in the group with perivascular hemorrhages (826 ± 394 µm) compared to the group with deep hemorrhages alone (455 ± 273 µm, P < 0.001). The 10 disc areas of retinal ischemia was more common in patients with perivascular (80.0%) and peripapillary (31.3%) versus deep hemorrhages alone (16.1%, P < 0.001). Neovascularization of the iris was more common, although this differrence was not significant, in the groups with peripapillary (14.3%) and perivascular (2.0%) NFL versus deep hemorrhages alone (0.0%). Conclusions: NFL retinal hemorrhages at baseline correlate with more severe forms of CRVO, with greater macular edema, poorer visual outcomes, and greater risk of ischemia and neovascularization. This may be related to the organization of the retinal capillary plexus. The depth and pattern of distribution of retinal hemorrhages in CRVO may provide an easily identifiable early biomarker of CRVO prognosis.

Neurological symptoms in patients with biopsy proven celiac disease.

In celiac disease (CD), the gut is the typical manifestation site but atypical neurological presentations are thought to occur in 6 to 10% with cerebellar ataxia being the most frequent symptom. Most studies in this field are focused on patients under primary neurological care. To exclude such an observation bias, patients with biopsy proven celiac disease were screened for neurological disease. A total of 72 patients with biopsy proven celiac disease (CD) (mean age 51 +/- 15 years, mean disease duration 8 +/- 11 years) were recruited through advertisements. All participants adhered to a gluten-free diet. Patients were interviewed following a standard questionnaire and examined clinically for neurological symptoms. Medical history revealed neurological disorders such as migraine (28%), carpal tunnel syndrome (20%), vestibular dysfunction (8%), seizures (6%), and myelitis (3%). Interestingly, 35% of patients with CD reported of a history of psychiatric disease including depression, personality changes, or even psychosis. Physical examination yielded stance and gait problems in about one third of patients that could be attributed to afferent ataxia in 26%, vestibular dysfunction in 6%, and cerebellar ataxia in 6%. Other motor features such as basal ganglia symptoms, pyramidal tract signs, tics, and myoclonus were infrequent. 35% of patients with CD showed deep sensory loss and reduced ankle reflexes in 14%. Gait disturbances in CD do not only result from cerebellar ataxia but also from proprioceptive or vestibular impairment. Neurological problems may even develop despite strict adherence to a gluten-free diet.

Changes in the OCT angiographic appearance of type 1 and type 2 CNV in exudative AMD during anti-VEGF treatment.

OBJECTIVE: Optical coherence tomography angiography (OCT-A) enables detailed visualisation of the vascular structure of choroidal neovascularisation (CNV). The aim of this study was to determine whether mathematically ascertained OCT-A vascular parameters of type 1 and type 2 CNV in exudative age-related macular degeneration (AMD) change during antivascular endothelial growth factor (anti-VEGF) treatment. The OCT-A vascular parameters were also compared with previously obtained activity parameters (fluid distribution on spectral domain OCT (SD-OCT)) to establish whether they could potentially be used as further 'activity parameters' for assessment of anti-VEGF treatment. METHODS AND ANALYSIS: We evaluated 27 eyes of 27 patients (mean follow-up 9.8 months) with type 1, type 2 or mixed CNV who had received anti-VEGF treatment (IVAN scheme). The parameters analysed were area (aCNV), total length of all vessels (tlCNV), overall number of vascular segments (nsCNV) and fractal dimension (FD) of the CNV. The changes in each of these parameters were correlated with the central foveal thickness (CFT). RESULTS: Regression and renewed perfusion of the CNV corresponded with the decrease or increase, respectively, of macular fluid distribution on SD-OCT. The increase and decrease of CFT during anti-VEGF treatment were highly significantly correlated with changes in FD (p<0.00001), aCNV (p<0.00001), tlCNV (p<0.00001) and nsCNV (p<0.00001). CONCLUSION: OCT-A enables detailed analysis of AMD with regard to FD, aCNV, tlCNV and nsCNV. As the changes in these parameters correlate closely with changes on SD-OCT, they can be used as new activity parameters, alongside fluid distribution, for assessment of treatment effect and as parameters of stabilisation or the need for repeated treatment.

Optical coherence tomography angiography of types 1 and 2 choroidal neovascularization in age-related macular degeneration during anti-VEGF therapy: evaluation of a new quantitative method.

PURPOSE: The aim of this pilot study was to test whether mathematical parameters of the vascular morphology of choroidal neovascularization (CNV) can be used as biomarkers and to investigate how these parameters change during anti-vascular endothelial growth factor (VEGF) therapy. METHODS: Treatment-naive CNV in exudative age-related macular degeneration (AMD) was diagnosed in 28 patients. OCT-angiography (OCT-A) (Avanti/FA Optovue) performed before and after anti-VEGF therapy. The OCT-A data sets were exported to an external image processing program and vessel skeletonization was accomplished by means of edge detection. Based on this technique the total vessel length, the number of segments and the fractal dimension (FD) of the CNV were calculated before and after therapy. The results were compared with other clinical parameters such as VA and central retinal thickness (RT). RESULTS: The total vessel length of the CNV was significantly reduced by anti-VEGF-therapy (mean value 652 pixels vs. 397 pixels; p < 0.0001), as well as the number of individual vessel segments of the CNV (mean value 117 vs. 76; p < 0.0001). The FD of the CNV also decreased significant reduction during therapy (mean 1.23 vs. 1.16, p < 0.0001). The changes in these parameters during treatment corresponded with an increase in VA and a reduction in RT. CONCLUSION: This pilot study demonstrates that the vascular pattern of CNV in AMD can be visualized and described using mathematical parameters of OCT-A. The changes during therapy correlate significantly with established "activity" parameters of CNV, so changes in these parameters (especially FD) may represent additional CNV "activity" biomarkers.

Negative Effects of Acid Violet-17 and MBB Dual In Vitro on Different Ocular Cell Lines.

PURPOSE: Vital dyes have become a clinical standard during vitrectomy to visualize anatomical structures. It was the aim of this study to test the effect of two vital dyes (AV 17-M with 5% mannitol and MBB Dual) on different intraocular cells, to see whether in vitro test can be used as reliable preclinical testing tool. METHODS: Cell morphology was assessed via phase contrast pictures, cell viability via MTS assay and total cell amount via crystal violet staining. ARPE19 and 661W cells were chosen for toxicology testing at different exposure times (60 seconds, 15 minutes and 30 minutes). Vital dyes were completely removed after the staining period. RESULTS: Treatment with AV 17-M changed the morphology and the cell number at every time point investigated on ARPE19 and 661 W cells. ARPE19 cells treated with AV 17-M or MBB Dual displayed only a slight or no decrease in cell viability after the three different exposure times. AV-17 without medium to simulate a possible intraoperative use after fluid-air exchange showed a decrease in viability of 6%, 24% and 14%. A difference in cell density of 21%, 46% and 34% was noted after CV staining for AV 17-M, MBB Dual led to a decrease of 2%, 16% and 3% after 30 minutes compared to BSS. AV 17-M directly applied on 661W decreased viability significantly by 18% after 60 seconds, 33% after 15 minutes and 40% after 30 minutes. Cell density of 661W cells exposed relevant negative effects; after incubation of 60 seconds with AV 17-M, the cell amount was significantly lowered by 41% and MBB Dual by 12%. After 15 minutes, a loss of 48% cell amount was detected with AV 17-M and after 30 min 51%. MBB Dual led to 37% loss after 15 minutes and to 28% loss after 30 minutes. CONCLUSION: AV 17-M with 5% mannitol has a negative effect on different ocular cells.