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Dog eye ulcer healing stages

Dog eye ulcer healing stages



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Dog eye ulcer healing stages using different wound dressings. The right lower quadrant of the eyes was injected with a 1% *N*-benzyl-[l]{.smallcaps}-cysteine solution (1 mL/5 g of eye, BD BBL, Franklin Lakes, NJ, USA) and incubated for 5 h. After washing the eyes with saline, 0.1 mL of a 1% formalin solution (10% buffered formalin) was injected into the eye socket, and the same volume of a fresh 1% formalin solution was injected on days 7, 14, and 21 after the acid injection. On day 0, 1, 2, 4, 7, 14, 21, and 28 after the acid injection, the acidified eyes were treated with the following wound dressings (as described in [Table 2](#tab2){ref-type="table"}).

4.4. Histological Analysis of Wound Healing {#sec4.4}

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On day 28 after the acid injection, the eye was removed from the animals, and the cornea was dissected to expose the wound area. The wounds were then excised and fixed in a 10% buffered formalin solution and were dehydrated in an ethanol series and embedded in paraffin. A 1 *μ*m section was stned with haematoxylin-eosin (H&,E) for histological analysis. Histological specimens were examined using a digital camera, and image analysis was performed using a computer system.

4.5. Measurement of Skin Wound Healing {#sec4.5}

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H&,E-stned specimens from the incision wounds in the anesthetized rats were prepared as described above and were photographed using a digital camera. Wound size was measured using Image-Pro Plus software (Media Cybernetics, Silver Spring, MD, USA).

4.6. Data Analysis {#sec4.6}

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All of the data were presented as the means ± SEM. Statistical analyses were performed using GraphPad Prism 5.0 (GraphPad Software, Inc., San Diego, CA, USA). The significant difference between the two groups was determined using one-way ANOVA. A *P* value <, 0.05 was considered significant.

This work was supported by the grants from the National Natural Science Foundation of China (81000580).

Conflict of Interests

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The authors declare that there is no conflict of interests regarding the publication of this paper.

![Acid-induced corneal surface changes. Representative cross-sections of cornea stned with H&,E on days 1, 3, 5, 7, 14, 21, and 28 after the acid injection.](ECAM2014-837892.001){#fig1}

![The distribution of acid-induced edema after 24 hours. The total area (a), average depth (b), maximum width (c), and minimum width (d) of the corneal edema from the limbus to the center of the cornea in four groups. **P* <, 0.05 versus saline. ^#^ *P* <, 0.05 versus control.](ECAM2014-837892.002){#fig2}

![The corneal morphology changes. Representative photomicrographs of cornea sections from each group on day 3 (a--d), day 5 (e--h), and day 7 (i--l) after acid injection. (a, b, e, f, i, and j) Normal cornea sections of control and LPC animals, (c, d, g, h, k, and l) corneal sections of LPC+RU animals, (a--d, e--h, and i--l) corneal sections of LPC+0.06% RU animals, (a--l) magnification ×4, (a--h) H&,E, (i--l) PAS.](ECAM2014-837892.003){#fig3}

![The corneal morphological changes from different groups after acid exposure. In saline control group, normal corneas were round (a), while central cornea were stellate-shaped (b), with normal corneal endothelial cells (c). LPC injury caused central corneal neovascularization (d). LPC injury combined with RU induced central corneal neovascularization (e) and endothelial cells apoptosis (f). LPC injury combined with 0.06% RU caused central corneal neovascularization and endothelial cells apoptosis (g), while normal endothelial cells are still found in the central corneal area (h). H&,E, bar = 40 *μ*m (a--h).](ECAM2014-837892.004){#fig4}

![Effect of ROS scavenging on corneal morphological changes. After acid exposure, corneas from the LPC-only, LPC+RU, and LPC+0.06% RU groups were fixed and analyzed on day 5 after injury. (a) Normal corneal morphology in control group. Central corneas were stellate-shaped (arrow). (b) Central corneal injury led to the infiltration of inflammatory cells (arrow). (c) Corneal neovascularization (arrow) was found in LPC-only treated animals. (d) 0.06% RU increased corneal neovascularization (arrow). (e) Central corneal neovascularization (arrow) was found in the LPC+0.06% RU group. H&,E, bar = 40 *μ*m (a--e).](ECAM2014-837892.005){#fig5}

![Effect of ROS scavenging on inflammatory cells in the injured cornea. (a) Inflammatory cells were found in corneas of LPC-only treated animals. (b) Inflammatory cells decreased in the LPC+0.06% RU group. (c) The number of inflammatory cells in the LPC+RU group was similar to that in the control group.](ECAM2014-837892.006){#fig6}

![Effect of ROS scavenging on corneal neovascularization in the injured cornea. (a) LPC-only treated group showed corneal neovascularization (arrow). (b) 0.06% RU was able to inhibit corneal neovascularization (arrow). (c) The corneal neovascularization was reduced by LPC+RU treatment.](ECAM2014-837892.007){#fig7}

![Effect of ROS scavenging on corneal opacity in the injured cornea. (a) Morphological changes of the injured cornea treated with LPC alone. (b) 0.06% RU was able to attenuate LPC-induced corneal opacity.](ECAM2014-837892.008){#fig8}

[^1]: Academic Editor: Josep M. Ribera


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