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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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Results of the terminal deoxynucleotidyl transferase-mediated nick end labeling assay. A: Apoptosis (red color) was observed only in a few cells in the normal cornea. B: In a cornea from group A, apoptosis was detected mainly in the posterior stroma and endothelial layer. C: In a cornea from group B, severe apoptosis throughout all layers of the cornea occurred. Scale bar=50 μm. *taken from the opposite eye that did not undergo cryoinjury.
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f6: Results of the terminal deoxynucleotidyl transferase-mediated nick end labeling assay. A: Apoptosis (red color) was observed only in a few cells in the normal cornea. B: In a cornea from group A, apoptosis was detected mainly in the posterior stroma and endothelial layer. C: In a cornea from group B, severe apoptosis throughout all layers of the cornea occurred. Scale bar=50 μm. *taken from the opposite eye that did not undergo cryoinjury.

Mentions: TUNEL assay of the samples taken one day after cryoinjury showed severe apoptosis throughout all corneal layers in group B, whereas apoptosis was mainly detected in the posterior stroma and endothelial layer in group A (Figure 6). The density of apoptotic cells in the cornea from group B was substantially higher than that in the cornea from group A, and the difference was statistically significant (433±168 cells/mm2 in group A versus 3167±798 cells/mm2 in group B, p<0.001).


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)

Results of the terminal deoxynucleotidyl transferase-mediated nick end labeling assay. A: Apoptosis (red color) was observed only in a few cells in the normal cornea. B: In a cornea from group A, apoptosis was detected mainly in the posterior stroma and endothelial layer. C: In a cornea from group B, severe apoptosis throughout all layers of the cornea occurred. 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

f6: Results of the terminal deoxynucleotidyl transferase-mediated nick end labeling assay. A: Apoptosis (red color) was observed only in a few cells in the normal cornea. B: In a cornea from group A, apoptosis was detected mainly in the posterior stroma and endothelial layer. C: In a cornea from group B, severe apoptosis throughout all layers of the cornea occurred. Scale bar=50 μm. *taken from the opposite eye that did not undergo cryoinjury.
Mentions: TUNEL assay of the samples taken one day after cryoinjury showed severe apoptosis throughout all corneal layers in group B, whereas apoptosis was mainly detected in the posterior stroma and endothelial layer in group A (Figure 6). The density of apoptotic cells in the cornea from group B was substantially higher than that in the cornea from group A, and the difference was statistically significant (433±168 cells/mm2 in group A versus 3167±798 cells/mm2 in group B, p<0.001).

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