The wound-healing response is critical to the outcome of refractive surgery and studying wound healing contributes to an understanding of the pathophysiology of other corneal injuries. Animal models allow research to be conducted with sufficient samples and under controlled parameters. We studied the hen to determine the healing process from clinical, biophysical, and biological standpoints after photorefractive keratectomy (PRK). PRK (ÿ6.0 diopters) was performed in hen eyes. At 3, 6, 12, 24, 48, and 72 h and 5, 7, 15, 30, and 60 days postoperatively, we studied the clinical follow-up, objective measurements of light transmission (direct transmittance), apoptosis by TUNEL assay, proliferation by immunocytochemical analysis of 5-bromo-2 0 -deoxyuridine, and expression of alpha smooth muscle actin (SMA) in myofibroblasts in the corneas. Hen corneas reepithelialize quickly. Haze developed from 5 to 60 days after surgery and was correlated with the appearance and finalization of the expression of SMA. The direct transmittance of light was low during the first 15 days and improved at 30 and 60 days. TUNEL-positive cells were observed 3 h after surgery and the numbers decreased thereafter. Epithelial proliferation began at 12 h and was greater at 48 h, while stromal cell proliferation began at 24 h and was greater at 72 h. The hen cornea is anatomically similar to the human cornea, and the manner in which it heals is a good model for studying different surgical techniques and pharmacologic assays.

The development of treatments that modulate corneal wound healing to avoid fibrosis during tissue repair is important for the restoration of corneal transparency after an injury. To date, few studies have studied the influence of growth factors (GFs) on human corneal fibroblast (HCF) expression of extracellular matrix (ECM) proteins such as collagen types I and III, proteoglycans such as perlecan, or proteins implicated in cellular migration such as α5β1-integrin and syndecan-4. Using in vitro HCFs, a mechanical wound model was developed to study the influence of the GFs basic fibroblast GF (bFGF), platelet-derived GF (PDGF-BB) and transforming GF-β1 (TGFβ1) on ECM protein production and cellular migration. Our results show that mechanical wounding provokes the autocrine release of bFGF and TGFβ1 at different time points during the wound closure. The HCF response to PDGF-BB was a rapid closure due to fast cellular migration associated with a high focal adhesion replacement and a high expression of collagen and proteoglycans, producing nonfibrotic healing. bFGF stimulated nonfibrotic ECM production and limited the migration process. Finally, TGFβ1 induced expression of the fibrotic markers collagen type III and α5β1 integrin, and it inhibited cellular migration due to the formation of focal adhesions with a low turnover rate. The novel in vitro HCF mechanical wound model can be used to understand the role played by GFs in human corneal repair. The model can also be used to test the effects of different treatments aimed at improving the healing process. Copyright © 2016 John Wiley & Sons, Ltd.

Purpose To evaluate corneal wound healing in the hen animal model after additive surgery with an intracorneal ring segment (ICRS). Methods We implanted one ICRS in each eye of 76 hens. In control group 1 (n=22 hens), the stromal channel was prepared but no ICRS was inserted. In control group 2 (n=2 hens), no surgery was performed. Animals were randomly separated into groups and euthanized after clinical follow-up of 4 and 12 hours, 1, 2, 3, and 7 days, and 1, 2, 3, 4, and 6 months. Corneas were stained with hematoxylin-eosin. Apoptosis was measured by terminal uridine nick end-labeling assays. Cell proliferation and myofibroblast-like differentiation were assayed by BrdU and α-smooth muscle actin immunofluorescence microscopy. Stromal matrix changes were documented by electron microscopy.Results Epithelial and stromal cell apoptosis around the ICRS-implanted and control group 1 eyes peaked at 12 hours, but continued for 72 hours. In ICRSimplanted eyes, epithelial and stromal proliferation was present at 12 and 24 hours, respectively, and peaked at 7 days and 72 hours, respectively. Some proliferation in the ICRS-implanted group continued through the 6-month follow-up, and myofibroblast-like cells differentiated one to three months after ICRS implantation. The segments rotated within the stroma as the limbal inferior angle approached the epithelium. Conclusions Wound healing after ICRS implantation in hen corneas was similar to that of other corneal surgical wounds in stages. However, there were some specific features related to the small size of the epithelial wound and the device permanently implanted inside the cornea.

The aim of this work is to provide experimental data for corneal transparency and scattering to help create a more complete model of corneal transparency. The scattered light in 96 healing hen corneas was measured for three wavelengths by a scatterometer constructed in the Optics Laboratory (The University of Valladolid, Spain). With the help of mirrors and beamsplitters, the light from the three lasers is directed toward the cell containing the sample to be measured. The measured scattered light varies between six orders of magnitude. Corneal transmissivity, mean cosine of a scattering angle, and angular distribution of scattered light were all computed. The total transmitted light remained practically constant over a wide range of light values transmitted in a forward direction (direct transmissivity). The value of the mean cosine of the scattering direction is very close to the unit (g 4 0:98), even in corneas with high opacities. The behavior of g indicates that even damaged corneas evidence extremely small scattering, compared to other biological tissues. The transmission reduction of each cornea is related to an increase in scattered light. In all cases, scattered light is concentrated at very small angles. This behavior is acceptable in corneas that are healthy or which evidence small lesions, but remains in corneas that are severely injured.

No abstract