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Published on: February 6, 2020
Longitudinal In Vitro Assessment of the Optical Transparency of Preserved Pre-Descemet Layer in the Absence of
Massimo Busin1,2,3, Angeli Christy Yu1,2,3, Vincenzo Scorcia4
1University of Ferrara, Department of Translational Medicine, Ferrara, Italy.
Purpose:
The purpose of this study was to evaluate the optical transparency morphological and ultrastructural features of the isolated pre-Descemet layer (PDL) in the absence of endothelium after preservation for up to 12 months using standard organ culture and silica gel-based tissue dehydration.
Methods:
Following removal of the Descemet membrane endothelium complex, the PDL was isolated using the big bubble technique. Ten samples were preserved either in standard organ culture medium (group 1) or by silica gel-based dehydration (group 2). Five samples with the PDL with thin posterior stroma served as the controls (group 3). Transparency was quantified using a custom-built optical device, and thickness was measured by anterior segment optical coherence tomography at baseline, 7 days, and 1, 3, 6, and 12 months. Ultrastructural analysis was then performed by transmission electron microscopy.
Results:
In group 1, mean transparency was 25.22 ± 4.32 lux (95% confidence interval [CI] = 19.87-30.57, SE = 1.93) at baseline and 19.13 ± 9.01 lux (95% CI = 7.94-30.32, SE = 4.03) at 12 months, with similar thickness (44.60 ± 8.42 µm (95% CI = 34.15-55.05, SE = 3.76) to 43.31 ± 8.69 µm (95% CI = 32.53-54.10, SE = 3.88). Group 2 showed comparable findings, with transparency of 20.9 ± 6.56 lux (95% CI = 12.75-29.04, SE = 2.93) at baseline and 19.37 ± 5.72 lux (95% CI = 12.26-26.47, SE = 1.56) after rehydration at 12 months, and thickness from 40.26 ± 7.73 µm (95% CI = 30.65-49.86, SE = 3.46) to 41.01 ± 8.18 µm (95% CI = 41.01-51.17, SE = 3.66). Control corneas (group 3) showed progressive loss of transparency (34.87 ± 6.56 lux, 95% CI = 26.72-43.01, SE = 2.93 to 43.4 ± 2.6 lux, 95% CI = 40.17-46.63, SE = 1.16) and stromal thickening (126.33 ± 33.25 µm (95% CI = 85.04-167.43, SE = 14.87) to 236.74 ± 61.98 µm (95% CI = 159.78-313.70, SE = 27.72). Electron microscopy revealed tightly packed collagen lamellae in groups 1 and 2 with similar appearance between preservation methods. Keratocytes were observed in group 3.
Conclusions:
Preserved PDL retains in vitro its transparency and thickness independent of endothelial function over 1 year.
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