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Alterations in proliferative activity in the corneal endothelial periphery after transcorneal freezing
Purpose To localise proliferating cells in the corneal endothelium after transcorneal freeze injury and detect possible progenitor cells. Methods Endothelial cells (ECs) were disrupted by transcorneal freezing in 5‐weeks‐old rabbits. Concomitantly 10 mg/kg 5‐Ethynyl‐2’‐deoxyuridine (EdU) was injecte...
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Published in: | Acta ophthalmologica (Oxford, England) England), 2016-10, Vol.94 (S256), p.n/a |
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Main Authors: | , , , , , , , , |
Format: | Article |
Language: | English |
Subjects: | |
Online Access: | Get full text |
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Summary: | Purpose
To localise proliferating cells in the corneal endothelium after transcorneal freeze injury and detect possible progenitor cells.
Methods
Endothelial cells (ECs) were disrupted by transcorneal freezing in 5‐weeks‐old rabbits. Concomitantly 10 mg/kg 5‐Ethynyl‐2’‐deoxyuridine (EdU) was injected intraperitoneally and repeated after 24 and 48 h. Animals were euthanized after 3, 5 or 40 days. Corneoscleral buttons were flat‐mounted for immunohistochemical analysis of proliferation marker Ki‐67 and detection of EdU, which is incorporated into the DNA during cell cycle S‐phase. Ki‐67+ and EdU+ cells were detected by fluorescence microscopy.
Results
Transcorneal freezing induced a central endothelial lesion of approximately 5 × 5 mm. After three days the endothelial wound was almost completely repopulated by small ECs exhibiting strong Ki‐67 expression and EdU incorporation. Furthermore, ECs in close proximity to the edges of the initial wound were positive for Ki‐67 and EdU. In the control group no central Ki‐67+ nor EdU+ ECs were detected. In the peripheral endothelium of the lesion group Ki‐67+ and EdU+ ECs were often arranged in clusters of 100–400 cells, with 1–3 clusters per quadrant. This is opposed to the periphery of the control group where Ki‐67+ and EdU+ ECs were abundantly present in all quadrants in a widespread pattern.
Conclusions
The central ECs in 5‐weeks‐old rabbits do not proliferate when the cell layer is intact, but proliferates extensively in response to wounding and covers the defect within 3 days. In contrast to this, peripheral ECs are continuously cycling. After a central lesion, the pattern of proliferative cells in the periphery change from widespread cells to clustered cells. This specific cell pattern can be indicative of a stem/progenitor cell niche. |
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ISSN: | 1755-375X 1755-3768 |
DOI: | 10.1111/j.1755-3768.2016.0632 |