Morphology of the pig cornea in normal conditions and after incubation in a perfusion apparatus

The fine structure of the pig cornea in normal conditions and after being used in a perfusion apparatus, for 4 h, is described. Earlier reports on the normal morphology of the pig cornea were partly not confirmed. Thus the number of cell layers in the epithelium was found to be 19-23 (a basal cell layer, 4–5 polyhedral cell layers and 14–17 squamous cell layers) compared to earlier reported 6–9 layers.The mean thickness of normal and perfused corneas were 722 μm and 752 μm respectively. Normal corneas had a hydration level of 77.2 % and after perfusion 78.5 %. The normal morphology and morphological changes due to exposure to perfusion were studied by light and electron microscopy. The differences observed between normal and perfused corneas have to be considered limited, and restricted mainly to the anterior squamous epithelium and the endothelium.Taken together our results indicate that the corneas used in the apparatus still had functional integrity.     

Pyogenic granuloma of the cornea in a pony: Uncommon complication of corneal wound healing

A 6-year-old miniature Shetland pony mare was referred for evaluation of a left corneal mass, which developed from the healing tissue of a corneal traumatic ulceration that had occurred 4 weeks previously. On gross examination, a spherical, smooth-surfaced, and pink-colored lesion of about 1 cm in diameter was protruding from the left palpebral fissure. Ophthalmic examination revealed that it was attached to the scar tissue of the cornea, and that one corpora nigra was adherent to the posterior face of corneal wounded area, without sign of uveitis. The remainder of the ophthalmic examination was unremarkable. The mass was excised, and cryotherapy was used as an adjunctive therapy. Histopathology of the resected mass was consistent with a pyogenic granuloma on the basis of radially oriented proliferating capillaries, embedded in immature granulation tissue containing an infiltrate of neutrophils, plasma cells and eosinophils. There were no histological features of malignancy. 2 months after surgery, the ventral part of the fibrotic corneal scar was slightly raised by a pink tissue, suggesting possible recurrence of the initial lesion. A second cryotherapy was performed over the leukoma area. No recurrence has been noted for a follow-up period of more than 25 months. Pyogenic granuloma is a benign proliferative fibrovascular response that typically develops after trauma or surgery. Corneal involvement is rare in humans, and to the authors' knowledge has never been documented in veterinary ophthalmology.     

Feline corneal disease

The cornea is naturally transparent. Anything that interferes with the cornea's stromal architecture, contributes to blood vessel migration, increases corneal pigmentation, or predisposes to corneal edema, disrupts the corneas transparency and indicates corneal disease. The color, location, and shape and pattern of a corneal lesion can help in determining the underlying cause for the disease. Corneal disease is typically divided into congenital or acquired disorders. Congenital disorders, such as corneal dermoids are rare in cats, whereas acquired corneal disease associated with nonulcerative or ulcerative keratitis is common. Primary ocular disease, such as tear film instability, adenexal disease (medial canthal entropion, lagophthalmus, eyelid agenesis), and herpes keratitis are associated with the majority of acquired corneal disease in cats. Proliferative/eosinophilic keratitis, acute bullous keratopathy, and Florida keratopathy are common feline nonulcerative disorders. Nonprogressive ulcerative disease in cats, such as chronic corneal epithelial defects and corneal sequestration are more common than progressive corneal ulcerations.     

Corneal sensitivity in healthy, immature, and adult alpacas

Objective To determine the corneal sensitivity of healthy, immature, and adult alpacas to establish normal reference values for this species. Animals studied Six normal crias and 18 normal adult alpacas. Procedures Corneal sensitivity was determined by evaluating the corneal touch threshold (CTT) in five regions of the cornea using a Cochet–Bonnet aesthesiometer. The nylon filament length in cm was then converted to applied pressure values in mg/0.0113 mm² and g/mm² using a conversion chart. Results The central region of the cornea was the most sensitive, and the dorsal and temporal regions were the least sensitive in alpacas. There were no significant differences between the right and left eyes or between values in males and females at any site. Crias had significantly greater corneal sensitivity compared with adult alpacas in all five regions of the cornea that were evaluated. Conclusions Normal reference values for CTT in alpacas were established using a Cochet–Bonnet aesthesiometer. The central region of the cornea is the most sensitive in alpacas, and crias have higher corneal sensitivity than adult alpacas.     

Treatment of corneal squamous cell carcinoma using topical 1% 5‐fluorouracil as monotherapy

The purpose of this report is to discuss the use of topical 1% 5‐fluorouracil as a sole therapy for canine corneal squamous cell carcinoma (SCC). A 12‐year‐old castrated male pug was evaluated for a well‐demarcated, central, 3 mm in diameter, pale pink, raised, right corneal mass. An incisional biopsy was obtained using a #64 beaver blade after topical anesthesia and without sedation. A definitive diagnosis of corneal SCC was obtained after histopathologic evaluation of the biopsy. Topical 1% 5‐fluorouracil ointment was applied to the right eye four times daily for 2 weeks followed by no treatment for 2 weeks, then treatment again twice daily for 2 weeks. The cornea remained free of recurrence 10 months after cessation of treatment. In dogs affected with corneal SCC, topical 1% 5‐fluorouracil monotherapy may be a viable and cost‐effective treatment option with minimal side effects. This chemotherapy agent may also have an effect on corneal pigmentation. Chronic cyclosporine therapy did not contribute to the pathogenesis of corneal SCC in the case described.     

Heterologous penetrating keratoplasty for treatment of a corneal sequestrum in a cat

A corneal sequestrum was diagnosed in an 8-year-old, neutered male Burmese cat. A heterologous penetrating keratoplasty (PK) (fresh canine corneal tissue) was performed to restore a clear visual axis. A heterograft was selected in order to decrease the risk of viral transmission as a screened donor was not available. One month postoperatively the graft was vascularized and opaque. The owner failed to return for recheck examinations until 16 months postoperatively at which time only a faint central nebula remained.     

Lamellar keratoplasty for the treatment of feline corneal sequestrum

A lamellar keratoplasty was used to treat corneal sequestrum in four Persian cats (six eyes). Following a superficial keratectomy, lamellar corneal allografts (feline corneal tissue) or heterografts (canine corneal tissue) which had been preserved at –20 °C were placed in the recipient cornea. All grafts became optically transparent within 2 months following surgery and no recurrences of the sequestrum have been noted during the follow-up period (4–30 months). We conclude that feline corneal sequestrum may be successfully treated with feline or canine donor corneal tissue using this technique.     

Early postnatal development of central corneal thickness in dogs

Objective To investigate the changes in corneal thickness that occur during maturation of the canine eye over the first months of life. Animals studied Dogs of two different breeds with ages ranging from 14 days to 42 weeks of age. Procedures The central corneal thickness was measured by ultrasonic pachymetry every week for the first month after eyelid opening (around 14 days) and then every month until 42 weeks of age. Segmented regression was applied to capture the two phases observed in the central corneal thickness plotted against age. Breed, eye and gender were also included in the model. Results Mean central corneal thickness (CCT) values initially decreased following eyelid opening, with the lowest point being reached at around 6 weeks of age. Then CCT gradually increased as the dogs matured. Differences between left and right eye were not significant. Breed and gender effects were significant factors in the statistical model. Conclusions Following eyelid opening there is an initial decrease in corneal thickness until approximately 6 weeks of age, which presumably mirrors maturation of corneal endothelial cell function. After 6 weeks of age the CCT increases with age until approximately 30 weeks of age after which there was only a gradual increase over the remainder of the study period. A similar pattern of changes in corneal thickness in humans has been previously recorded.     

Cultivation of corneal epithelial cell sheets on canine amniotic membrane

Objective To develop and assess canine corneal epithelial cell sheets cultivated from limbal stem cells on amniotic membrane. Procedures Canine corneal limbal segments were obtained from six beagle dogs. Cryopreserved denuded amniotic membranes (obtained from Miniature Dachshund and Cavalier King Charles Spaniel breeds) from which the epithelial cells were removed were used as scaffolds. The limbal segments were cultured on these amniotic membranes with 3T3 feeder cells for 2 weeks. The harvested corneal epithelial cell sheets were stained with H&E for histologic analysis. The harvested sheets were analyzed immunohistochemically using a corneal epithelium‐specific marker keratin 3(K3) and putative stem cell markers ABCG2, p63, and vimentin. Results Cultivated cells from the corneal limbal tissues reached confluency in 7–8 days. The cultivated cells adhered to the denuded amniotic membrane and formed a sheet. The cultivated cell sheet was transparent and consisted of five to eight layers. K3 was observed in all layers and ABCG2, p63, and vimentin were notably present in the basal layer of the cultivated canine epithelium by immunofluorescence. Conclusions Canine corneal epithelial cells were successfully cultivated on the canine amniotic membrane. The cultivated epithelial sheets contained putative stem cells in the basal layer and had a stratified epithelium.