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LIM Homeobox Domain 2 Is Required for Corneal Epithelial Homeostasis.

Sartaj R, Chee RI, Yang J, Wan P, Liu A, Guaiquil V, Fuchs E, Rosenblatt MI - Stem Cells (2016)

Bottom Line: Immunodetection on corneal sections were used to visualize conjunctivalization, a sign of limbal barrier failure.Cell based assays showed that Lhx2cKO derived corneal epithelial cells have a significantly lower capacity to form colonies over time and delayed wound-healing recovery when compared to wildtype cells.We conclude that Lhx2 is required for maintenance of the corneal epithelial cell compartment and the limbal barrier.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, University of Illinois at Chicago, Chicago, Illinois, USA.

No MeSH data available.


Related in: MedlinePlus

Corneal epithelial wound healing is impaired in Lhx2cKO mice.We assessed the renewal capacity of the corneal epithelium in WT and Lhx2cKO mice by inducing consecutive epithelial injuries and visualizing the area of the debridement by fluorescein staining. The mice were subjected to cornea epithelial debridement three times, allowing a week to recover between each procedure (see Materials and Methods). In general, WT mice healed their epithelium consistently at 32hrs post‐injury, whilst a significant subset of Lhx2cKO mice demonstrated markedly delayed wound closure. After a third epithelial debridement, WT mice healed their epithelium at 32hrs while most Lhx2cKO had persistent epithelial defects at 144 hrs post wounding (typical example of this marked delay in healing shown) (A). A detailed graphical representation of all epithelial debridements showed that Lhx2cKO mice (far right panel) have a reduced healing capacity after repeated debridement and presented corneal complications, such as corneal neovascularization and perforation that were not observed in the WT (far left panel) (B). Abbreviation: WT, wild type.
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stem2257-fig-0005: Corneal epithelial wound healing is impaired in Lhx2cKO mice.We assessed the renewal capacity of the corneal epithelium in WT and Lhx2cKO mice by inducing consecutive epithelial injuries and visualizing the area of the debridement by fluorescein staining. The mice were subjected to cornea epithelial debridement three times, allowing a week to recover between each procedure (see Materials and Methods). In general, WT mice healed their epithelium consistently at 32hrs post‐injury, whilst a significant subset of Lhx2cKO mice demonstrated markedly delayed wound closure. After a third epithelial debridement, WT mice healed their epithelium at 32hrs while most Lhx2cKO had persistent epithelial defects at 144 hrs post wounding (typical example of this marked delay in healing shown) (A). A detailed graphical representation of all epithelial debridements showed that Lhx2cKO mice (far right panel) have a reduced healing capacity after repeated debridement and presented corneal complications, such as corneal neovascularization and perforation that were not observed in the WT (far left panel) (B). Abbreviation: WT, wild type.

Mentions: Corneal injury is repaired from proliferating corneal epithelial cells, originating from the limbus. Re‐epithelialization can be visualized during this process by fluorescein that stains the cornea devoid of epithelium. To determine the role of Lhx2 in maintaining the stemness capacity of the limbus, we challenged the corneal epithelium with multiple epithelial debridements to establish if the re‐epithelialization of the injured cornea proceeds in a similar fashion in Lhx2cKO and WT mice. The mice (WT n = 8, Lhx2cKO n = 9) were subjected to epithelial debridement repeatedly once a week for 3 weeks. We used the right eyes for debridement and the contralateral eye as a control. We found that Lhx2cKO mice have impaired capacity to heal the injured corneal epithelium. Slit lamp examination demonstrated that WT corneas routinely healed by 32 hours after a third debridement, whereas Lhx2cKO corneas had a marked delay in re‐epithelialization with only 30% of these corneas healed at 144 hours (Fig. 5A). Experimental data from epithelial debridement 1, 2, and 3 from WT (left) and Lhx2cKO (right) are collectively represented in Figure 5B. We registered the epithelial wound healing response found in the cornea of both WT and Lhx2cKO after these series of debridements. We observed no phenotypic alteration in the cornea of WT mice in any of the experiments. WT mice showed faster epithelial wound healing in the second and third debridement when compared with the initial first debridement (Fig. 5B). However, in the corneas of Lhx2cKO mice, we found that some mice healed within the normal time frame after the first debridement and some took longer to heal. After the second and third debridement, we observed an increase in the number of mice that showed epithelial defects or did not heal (Fig. 5B). The epithelial defects usually started with cornea neovascularization (white squares) that lead to perforation (red squares), keratinization, opacification, and loss of vision.


LIM Homeobox Domain 2 Is Required for Corneal Epithelial Homeostasis.

Sartaj R, Chee RI, Yang J, Wan P, Liu A, Guaiquil V, Fuchs E, Rosenblatt MI - Stem Cells (2016)

Corneal epithelial wound healing is impaired in Lhx2cKO mice.We assessed the renewal capacity of the corneal epithelium in WT and Lhx2cKO mice by inducing consecutive epithelial injuries and visualizing the area of the debridement by fluorescein staining. The mice were subjected to cornea epithelial debridement three times, allowing a week to recover between each procedure (see Materials and Methods). In general, WT mice healed their epithelium consistently at 32hrs post‐injury, whilst a significant subset of Lhx2cKO mice demonstrated markedly delayed wound closure. After a third epithelial debridement, WT mice healed their epithelium at 32hrs while most Lhx2cKO had persistent epithelial defects at 144 hrs post wounding (typical example of this marked delay in healing shown) (A). A detailed graphical representation of all epithelial debridements showed that Lhx2cKO mice (far right panel) have a reduced healing capacity after repeated debridement and presented corneal complications, such as corneal neovascularization and perforation that were not observed in the WT (far left panel) (B). Abbreviation: WT, wild type.
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stem2257-fig-0005: Corneal epithelial wound healing is impaired in Lhx2cKO mice.We assessed the renewal capacity of the corneal epithelium in WT and Lhx2cKO mice by inducing consecutive epithelial injuries and visualizing the area of the debridement by fluorescein staining. The mice were subjected to cornea epithelial debridement three times, allowing a week to recover between each procedure (see Materials and Methods). In general, WT mice healed their epithelium consistently at 32hrs post‐injury, whilst a significant subset of Lhx2cKO mice demonstrated markedly delayed wound closure. After a third epithelial debridement, WT mice healed their epithelium at 32hrs while most Lhx2cKO had persistent epithelial defects at 144 hrs post wounding (typical example of this marked delay in healing shown) (A). A detailed graphical representation of all epithelial debridements showed that Lhx2cKO mice (far right panel) have a reduced healing capacity after repeated debridement and presented corneal complications, such as corneal neovascularization and perforation that were not observed in the WT (far left panel) (B). Abbreviation: WT, wild type.
Mentions: Corneal injury is repaired from proliferating corneal epithelial cells, originating from the limbus. Re‐epithelialization can be visualized during this process by fluorescein that stains the cornea devoid of epithelium. To determine the role of Lhx2 in maintaining the stemness capacity of the limbus, we challenged the corneal epithelium with multiple epithelial debridements to establish if the re‐epithelialization of the injured cornea proceeds in a similar fashion in Lhx2cKO and WT mice. The mice (WT n = 8, Lhx2cKO n = 9) were subjected to epithelial debridement repeatedly once a week for 3 weeks. We used the right eyes for debridement and the contralateral eye as a control. We found that Lhx2cKO mice have impaired capacity to heal the injured corneal epithelium. Slit lamp examination demonstrated that WT corneas routinely healed by 32 hours after a third debridement, whereas Lhx2cKO corneas had a marked delay in re‐epithelialization with only 30% of these corneas healed at 144 hours (Fig. 5A). Experimental data from epithelial debridement 1, 2, and 3 from WT (left) and Lhx2cKO (right) are collectively represented in Figure 5B. We registered the epithelial wound healing response found in the cornea of both WT and Lhx2cKO after these series of debridements. We observed no phenotypic alteration in the cornea of WT mice in any of the experiments. WT mice showed faster epithelial wound healing in the second and third debridement when compared with the initial first debridement (Fig. 5B). However, in the corneas of Lhx2cKO mice, we found that some mice healed within the normal time frame after the first debridement and some took longer to heal. After the second and third debridement, we observed an increase in the number of mice that showed epithelial defects or did not heal (Fig. 5B). The epithelial defects usually started with cornea neovascularization (white squares) that lead to perforation (red squares), keratinization, opacification, and loss of vision.

Bottom Line: Immunodetection on corneal sections were used to visualize conjunctivalization, a sign of limbal barrier failure.Cell based assays showed that Lhx2cKO derived corneal epithelial cells have a significantly lower capacity to form colonies over time and delayed wound-healing recovery when compared to wildtype cells.We conclude that Lhx2 is required for maintenance of the corneal epithelial cell compartment and the limbal barrier.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, University of Illinois at Chicago, Chicago, Illinois, USA.

No MeSH data available.


Related in: MedlinePlus