Abstract
P 078
Esterified-cholesterol containing substructures in extra-macular drusen of human Bruch’s membrane wholemounts
Katja Seckerdieck1, Christine A. Curcio2, Salvatore Grisanti1, Martin Rudolf1
1Klinik für Augenheilkunde, Campus Lübeck, Universitätsklinikum Schleswig-Holstein, Lübeck, 2Department of Ophthalmology, Callahan Eye Foundation Hospital, University of Alabama at Birmingham
Objective
To evaluate the distribution of esterified cholesterol, an important druse constituent in extra-macular drusen in wholemount (WM) preparations of Bruch’s membrane (BrM).
Methods
BrM-choroid of 3 donor eyes (76-86 years) fixed in 4% paraformaldehyde were prepared including cautious RPE-removal. WMs were stained with filipin for esterified cholesterol and extra-macular drusen were evaluated by wide-field epifluorescence microscopy with parameters previously used for drusen in histological sections by.
Results
A total of 356 drusen were judged: The median of the drusen size was 24.85 µm (range 8.24-93.77 µm), 147 drusen (41.3%) demonstrated an intense and evenly distributed fluorescence throughout the entire druse (full body staining); 67 (18.8%) showed a hypoflourescent core (range 3.24-18.97 µm), 10 large drusen (2.8%) had more than 1 core (probably after advanced confluence); 37 drusen (10.4%) had shells; 186 drusen (52.2%) had lakes (comma-shaped hyperfluorescent areas), abundantly in 108 drusen (30,3%) and scattered in 78 drusen (21.9%). We found clusters containing up to 7 merging drusen. It is noteworthy that all merging drusen of a group exhibited almost identical staining characteristics.
Conclusions
The WM perspective reveals more information about cholesterol deposition in drusen, because sample size is large and an entire drusen can be seen as well neighbours. Our data on druse substructure prevalence differs somewhat from our previous data obtained from sections of extra-macular drusen, as we found esterified cholesterol-rich lakes more frequently and shells less frequently than previously. Judgement of drusen confluence is more reliable in WMs than in sections. All merging drusen of a cluster exhibited the same staining pattern which might indicate that local mechanisms might initiate certain sequences of drusen biogenesis.
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