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A mouse model of corneal endothelial decompensation using cryoinjury.

Han SB, Ang H, Balehosur D, Peh G, Chaurasia SS, Tan DT, Mehta JS - Mol. Vis. (2013)

Bottom Line: IOP remained within normal range in group A, but increased significantly with time in group B (p=0.011 at day 1, 0.038 at day 3, 0.026 at day 14, and 0.008 at day 21).Live/dead cell assay, hematoxylin and eosin staining, and scanning electron microscopy revealed successful ablation of corneal endothelial cells and absence of regeneration in both groups.One cycle of cryoinjury was safer than three, while both were equally effective in inducing bullous keratopathy.

View Article: PubMed Central - PubMed

Affiliation: Singapore National Eye Centre, Singapore.

ABSTRACT

Purpose: To develop a mouse model of bullous keratoplasty and evaluate the safety and efficacy of cryoinjury-induced corneal endothelial decompensation.

Methods: Transcorneal freezing was performed on the right eye of each mouse. One cycle of cryoinjury was performed in 18 eyes (group A), and three cycles were performed in 17 eyes (group B). Pachymetry and intraocular pressure (IOP) measurements were done preoperatively, as well as at 1, 3, 7, 14, and 21 days after cryoinjury. At each post-cryoinjury time point, three mice from each group were euthanized, and the corneas underwent histology and electron microscopy.

Results: In both groups, significant corneal edema was noted at post-cryoinjury day 1, which was maintained throughout the study period. IOP remained within normal range in group A, but increased significantly with time in group B (p=0.011 at day 1, 0.038 at day 3, 0.026 at day 14, and 0.008 at day 21). In group B, serious complications including hyphema (one case), severe iridocorneal adhesion (15 cases), and total cataract (three cases) were detected, while only mild iridocorneal adhesion (four cases) and cataract (three cases) were noted in group A. Live/dead cell assay, hematoxylin and eosin staining, and scanning electron microscopy revealed successful ablation of corneal endothelial cells and absence of regeneration in both groups. Hematoxylin and eosin staining and terminal deoxynucleotidyl transferase-mediated nick end labeling assay showed that apoptosis was mainly confined to the posterior stroma and endothelium in group A, while severe apoptosis was observed throughout all layers of the cornea in group B.

Conclusions: One cycle of cryoinjury was safer than three, while both were equally effective in inducing bullous keratopathy. This cryoinjury mouse model of bullous keratopathy was a consistently reproducible model that can be used for further studies on endothelial cell damage and rescue therapy.

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Related in: MedlinePlus

Live/dead cell assay using alizarin red S and Trypan Blue staining of the corneal endothelial cells. Successful ablation of the corneal endothelial cells was observed in samples from day 1, and no evidence of cell regrowth was detected during the study period. Descemet’s membrane was preserved over most of the area. Scale bar=50 μm. *taken from the opposite eye that did not undergo cryoinjury.
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f4: Live/dead cell assay using alizarin red S and Trypan Blue staining of the corneal endothelial cells. Successful ablation of the corneal endothelial cells was observed in samples from day 1, and no evidence of cell regrowth was detected during the study period. Descemet’s membrane was preserved over most of the area. Scale bar=50 μm. *taken from the opposite eye that did not undergo cryoinjury.

Mentions: In both groups, staining of CECs with alizarin red S and trypan blue showed complete ablation of the corneal endothelial layer and only small clusters of remnant CECs were noted in the periphery (Figure 4). There was no evidence of repopulation of CECs with time noted in either group. In samples from all the time points, DM was mostly preserved, although defects were found in some areas. There appeared to be no significant difference in the extent of DM damage between the two groups and among the various time points.


A mouse model of corneal endothelial decompensation using cryoinjury.

Han SB, Ang H, Balehosur D, Peh G, Chaurasia SS, Tan DT, Mehta JS - Mol. Vis. (2013)

Live/dead cell assay using alizarin red S and Trypan Blue staining of the corneal endothelial cells. Successful ablation of the corneal endothelial cells was observed in samples from day 1, and no evidence of cell regrowth was detected during the study period. Descemet’s membrane was preserved over most of the area. Scale bar=50 μm. *taken from the opposite eye that did not undergo cryoinjury.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3675054&req=5

f4: Live/dead cell assay using alizarin red S and Trypan Blue staining of the corneal endothelial cells. Successful ablation of the corneal endothelial cells was observed in samples from day 1, and no evidence of cell regrowth was detected during the study period. Descemet’s membrane was preserved over most of the area. Scale bar=50 μm. *taken from the opposite eye that did not undergo cryoinjury.
Mentions: In both groups, staining of CECs with alizarin red S and trypan blue showed complete ablation of the corneal endothelial layer and only small clusters of remnant CECs were noted in the periphery (Figure 4). There was no evidence of repopulation of CECs with time noted in either group. In samples from all the time points, DM was mostly preserved, although defects were found in some areas. There appeared to be no significant difference in the extent of DM damage between the two groups and among the various time points.

Bottom Line: IOP remained within normal range in group A, but increased significantly with time in group B (p=0.011 at day 1, 0.038 at day 3, 0.026 at day 14, and 0.008 at day 21).Live/dead cell assay, hematoxylin and eosin staining, and scanning electron microscopy revealed successful ablation of corneal endothelial cells and absence of regeneration in both groups.One cycle of cryoinjury was safer than three, while both were equally effective in inducing bullous keratopathy.

View Article: PubMed Central - PubMed

Affiliation: Singapore National Eye Centre, Singapore.

ABSTRACT

Purpose: To develop a mouse model of bullous keratoplasty and evaluate the safety and efficacy of cryoinjury-induced corneal endothelial decompensation.

Methods: Transcorneal freezing was performed on the right eye of each mouse. One cycle of cryoinjury was performed in 18 eyes (group A), and three cycles were performed in 17 eyes (group B). Pachymetry and intraocular pressure (IOP) measurements were done preoperatively, as well as at 1, 3, 7, 14, and 21 days after cryoinjury. At each post-cryoinjury time point, three mice from each group were euthanized, and the corneas underwent histology and electron microscopy.

Results: In both groups, significant corneal edema was noted at post-cryoinjury day 1, which was maintained throughout the study period. IOP remained within normal range in group A, but increased significantly with time in group B (p=0.011 at day 1, 0.038 at day 3, 0.026 at day 14, and 0.008 at day 21). In group B, serious complications including hyphema (one case), severe iridocorneal adhesion (15 cases), and total cataract (three cases) were detected, while only mild iridocorneal adhesion (four cases) and cataract (three cases) were noted in group A. Live/dead cell assay, hematoxylin and eosin staining, and scanning electron microscopy revealed successful ablation of corneal endothelial cells and absence of regeneration in both groups. Hematoxylin and eosin staining and terminal deoxynucleotidyl transferase-mediated nick end labeling assay showed that apoptosis was mainly confined to the posterior stroma and endothelium in group A, while severe apoptosis was observed throughout all layers of the cornea in group B.

Conclusions: One cycle of cryoinjury was safer than three, while both were equally effective in inducing bullous keratopathy. This cryoinjury mouse model of bullous keratopathy was a consistently reproducible model that can be used for further studies on endothelial cell damage and rescue therapy.

Show MeSH
Related in: MedlinePlus