In vivo confocal microscopy of the normal equine cornea and limbus

Objective  To describe morphologic features, pachymetry and endothelial cell density of the normal equine cornea and limbus by in vivo confocal microscopy.
Animals studied  Ten horses without ocular disease.
Procedure  The central and peripheral corneas were examined with a modified Heidelberg Retina Tomograph II and Rostock Cornea Module using a combination of automated and manual image acquisition modes. Thickness measurements of various corneal layers were performed and endothelial cell density determined.
Results  Images of the constituent cellular and noncellular elements of the corneal epithelium, stroma, endothelium, and limbus were acquired in all horses. Corneal stromal nerves, the subepithelial nerve plexus, and the sub-basal nerve plexus were visualized. Cells with an appearance characteristic of Langerhans cells and corneal stromal dendritic cells were consistently detected in the corneal basal epithelium and anterior stroma, respectively. Median central total corneal thickness was 835 μm (range 725–920 μm) and median central corneal epithelial thickness was 131 μm (range 115–141 μm). Median central endothelial cell density was 3002 cells per mm² (range 2473–3581 cells per mm²).
Conclusions  In vivo corneal confocal microscopy provides a noninvasive method of assessing normal equine corneal structure at the cellular level and is a precise technique for corneal sublayer pachymetry and cell density measurements. A resident population of presumed Langerhans cells and corneal stromal dendritic cells was detected in the normal equine cornea. The described techniques can be applied to diagnostic evaluation of corneal alternations associated with disease and have broad clinical and research applications in the horse.     

In vivo confocal microscopy in the normal corneas of cats, dogs and birds

OBJECTIVE: To evaluate the applicability of in vivo confocal microscopy (IVCM) in veterinary ophthalmology and analyze the morphology of living, healthy cornea. ANIMALS EXAMINED: Thirty-seven dogs, 34 cats and five birds. PROCEDURE: Various corneal sublayers were visualized in the central region using an in vivo confocal corneal microscope (HRTII/RCM). RESULTS: An investigation method was developed and adapted for use on animals with varying skull forms and eye positions. Real-time images of the epithelial cells, the corneal stroma and the endothelial layer were obtained. The corneal stromal nerve trunks and the subepithelial and basal epithelial nerve plexus were visualized. In dogs, full corneal thickness (FCT) was 585 +/- 79 microm (mean +/- SD) and endothelial cell density (ECD) 3175 +/- 776 cells/mm(2) (mean +/- SD). In cats, FCT was 592 +/- 80 microm and ECD 2846 +/- 403 cells/mm(2). There were no significant differences between canine and feline FCT and ECD and no morphologic differences could be seen between dogs and cats. The bird images revealed a number of structural differences. CONCLUSION: Noninvasive IVCM allows accurate detection of corneal sublayers, corneal pachymetry, endothelial cell density and corneal innervation in various animal species. For clinical usage, patients must be under general anesthesia. The confocal images provided anatomic reference images of various healthy corneal structures in dogs, cats and birds.     

A cross‐linked hyaluronan gel accelerates healing of corneal epithelial abrasion and alkali burn injuries in rabbits

Objective To evaluate the efficacy of a chemically modified and cross‐linked derivative of hyaluronan (CMHA‐SX) for treatment of corneal epithelial abrasion and standardized alkali burn injuries. Animals Twelve female New Zealand white rabbits in two groups were used. Procedures Bilateral 6‐mm diameter corneal epithelial abrasions were made in each of six rabbits in one group and 6‐mm standardized alkali burn injuries were made in the second group. A 1% CMHA‐SX formulation was applied topically four times per day in right eye of each rabbit for 1 week, and phosphate buffered saline (PBS) was placed in left (control) eye of each rabbit. The wound size was determined by staining with 1% fluorescein and photographed at the slit lamp with a digital camera at 0, 1, 2, 3 days postoperatively in the first group and 0, 1, 2, 3, 7, 12 days in the second group. Rabbit corneas were collected for histological examination on day 7 in the first group and day 12 in the second group. Results Closure of corneal wound in the abrasion model was complete in the CMHA‐SX treated eye by 48 h. The wound closure rate and thickness of the central corneal epithelium in the CMHA‐SX treated group was greater than in control eyes for both the abrasion and alkali burn injuries. Moreover, the CMHA‐SX treated cornea exhibited better epithelial and stromal organization than the untreated control cornea. Conclusions Chemically modified and cross‐linked derivative of hyaluronan improved corneal wound healing and could be useful for treating noninfectious corneal injuries.     

Reconstruction of corneal epithelium with cryopreserved corneal limbal stem cells in a rabbit model

The integrity and transparency of the cornea plays a key role in preserving vision. This paper reports a procedure to create an artificial sheet of corneal epithelium from cryopreserved limbal stem cells (LSCs) and to use this for corneal transplantation. Corneal LSCs were isolated from biopsy specimens of rabbit limbal lamellar and cryopreserved in liquid nitrogen at 2–4 passages. The cells were grown in culture medium for 12–14 days on top of a cell-free human amniotic membrane framed on a nitrocellulose sheet. The corneal epithelium generated was transplanted into the right eyes of 14 LSC deficient (LSCD) rabbits (seven experimental animals, seven controls) with corneal damage. The seven LSCD rabbits in the experimental group were transplanted with a corneal epithelial sheet generated from the cryopreserved corneal LSCs. Four LSCD rabbits were used as the vehicle control and were transplanted with a cell-free amniotic membrane, and the remaining three LSCD rabbits were negative controls without transplantation. Over a 2-month recovery period, 2/7 animals in the experimental group recovered completely, four recovered partially and one did not respond. In the control groups, three negative controls and three vehicle controls lost their vision completely, and one of the vehicle controls partially recovered transparency of the cornea Following treatment, corneal transparency of the experimental rabbits was significantly improved compared to controls (P < 0.05). The results indicated that cryopreserved corneal LSCs can repair damaged rabbit cornea, suggesting a possible new clinical approach to reconstruction of corneal epithelium.     

Canine corneal thickness measured by ultrasonic pachymetry.

Ultrasonic pachymetry was used to measure central, superior peripheral, and temporal peripheral corneal thicknesses of 75 dogs (150 eyes) with normal corneas, anterior chambers, and intraocular pressure. Mean corneal thickness averaged over the 2 eyes, 3 locations, and 75 dogs was 562 +/- 6.2 microns. The peripheral cornea was thicker on average than the central cornea by 49.43 +/- 8.45 microns and this difference increased with age at 6.97 +/- 1.3 microns/month of age. Mean corneal thickness changed with age (14.23 +/- 2.26 microns/month), and weight (1.83 +/- 0.38 microns/kg). Females had significantly thinner corneas (22.43 +/- 11.03 microns than males) after adjusting for age and weight.     

Corneal diagnostic procedures

The cornea is the anterior, transparent portion of the fibrous tunic of the eye. It is continuous with the sclera at a transition called the limbus. In healthy conditions, the transparency of the cornea is maintained by the smooth, nonkeratinized, squamous epithelium, which is further enhanced by the precorneal tear film, the lack of corneal vascularization or pigmentation, the size and regular arrangement of the collagen fibrils that make up the corneal stroma, and the relative dehydration of the cornea (which is maintained by the endothelium and epithelium). The cornea can respond to adverse stimuli through vascularization, pigmentation, fibrosis, accumulation of cellular or noncellular infiltrate, and/or edema. Because of these limited responses, routine diagnostic procedures are critical in the diagnosis and treatment of corneal disorders. This article discusses tests of the precorneal tear film, corneal staining procedures, culture and sensitivity, cytology, and a few other procedures that are performed less commonly or require specialized instrumentation.     

Major histocompatibility class II expression in the normal canine cornea and in canine chronic superficial keratitis

OBJECTIVE: To compare the expression of major histocompatibility complex (MHC) class II antigen in the corneas of normal dogs and dogs affected with chronic superficial keratitis (CSK). METHODS: MHC class II expression was determined in frozen sections of normal canine cornea and cornea from lesions of CSK by immunohistochemistry using a monoclonal antibody directed against the canine MHC class II molecule. Langerhans cell phenotype was determined morphologically and by histochemical determination of ATPase activity. To determine the influence of gamma interferon on expression of MHC class II molecules by corneal cells, corneal explants were cultured with the cytokine and MHC class II expression determined as above. RESULTS: Numerous MHC class II-expressing cells were demonstrated within the stroma and epithelium of the normal corneal limbus and conjunctival epithelium while very little MHC class II expression was detected in the central region of normal canine cornea. In limbal and conjunctival epithelium, cells expressing MHC class II antigen showed ATPase activity, suggesting that they were Langerhans cells. Corneas from dogs with CSK showed MHC class II expression associated with stromal cells, some of which exhibited a dendritic morphology while most were lymphocytic. Corneal epithelial cells within the lesion also aberrantly expressed MHC class II. Corneal explants expressed MHC class II to varying degrees after differing periods of incubation with the cytokine gamma interferon. CONCLUSIONS: While the normal central cornea has little MHC class II expression, aberrant expression occurs in CSK, associated with secretion of gamma interferon by infiltrating CD4-expressing lymphocytes. Although this change is likely to be a secondary feature of the CSK lesion, increased MHC class II expression may play a part in perpetuating the corneal inflammation seen in the disease.     

Assessment of the posterior segment of the cat eye by optical coherence tomography (OCT)

OBJECTIVES: To assess the feasibility of optical coherence tomography (OCT) for examining the cat ocular fundus, to provide normative data on retinal thickness in different fundus regions, and to demonstrate selected surgically induced vitreoretinal pathologies in the cat. ANIMAL STUDIED: Forty-five eyes of 28 healthy domestic cats and two eyes of domestic cats that had undergone subretinal implantation surgery for a visual prosthesis were examined. PROCEDURES: An optical coherence tomograph (Zeiss-Humphrey) was used to examine the anesthetized animals. At least five vertical and five horizontal scans in regular distribution were recorded for each cat including (1) the peripapillary region, (2) the area centralis, and (3) the peripheral retina. Thickness was measured manually at five locations in each scan. Retinal thickness was compared in the three above-mentioned fundus regions, between eyes and between vertical and horizontal scans. OCT was additionally performed in animals with retinal detachment and a subretinal visual prosthesis. RESULTS: OCT measurements required only minimal adjustments of human settings and yielded high quality images. In comparison to humans intraretinal layers were more difficult to differentiate. Retinal thickness was highest in the peripapillary region (245 +/- 21 microm), followed by the peripheral retina (204 +/- 11 microm) and the area centralis (182 +/- 11 microm; all P < 0.0001). There was no statistically significant difference between right and left eye or between vertical and horizontal scans. OCT demonstrated retinal detachment, an iatrogenic break and a subretinal prosthetic device in high detail. CONCLUSIONS: Retinal thickness was measurable with high precision; values compare well to older histologic studies. OCT bears significant advantages over histology in enabling one to repeat measurements in living animals and thus allowing longitudinal studies. Various vitreoretinal pathologies common in feline eyes are detectable and quantifiable by OCT.     

Normal corneal thickness measurements in pigmented rabbits using spectral‐domain anterior segment optical coherence tomography

Objective  To document the thickness of the central cornea in pigmented rabbits using spectral‐domain anterior segment optical coherence tomography (AS‐OCT). Animals studied  Seventeen pigmented rabbits (6 male, 11 female, both eyes) were involved in this study. Procedures  Thirty‐four eyes from healthy pigmented rabbits underwent a complete ophthalmologic examination, including AS‐OCT. Eight radial scans, 6 mm in length and centered on the cornea, were obtained using the AS‐OCT. Corneal thickness was automatically calculated using pachymetry software. Measurements were displayed as the mean and standard deviation for each of the 17 regions defined by the software. The regions were the center (1 mm radius, area a), the inner ring (2.5 mm radius, area b), the outer ring (3 mm radius, area c), and the eight radial scan lines in eight directions (Superior (1), SN (2), Nasal (3), IN (4), Inferior (5), IT (6), Temporal (7), ST (8)) with an angle of 45° between each consecutive scan line (a, b 1–8, c 1–8). Results  There was no statistically significance difference in corneal thickness between gender, eye, and the eight directions (P = 0.804, P > 0.05, P > 0.05). There was a statistical difference between the thickness in areas a, b, and c (P < 0.05). The corneal thickness increased gradually from the center to the periphery of the 6 mm measured. The center corneal thickness was 387 ± 19.8 μm for OD and 384 ± 20.2 μm for OS. The corneal thicknesses of the thinnest point of the right eyes (OD) and the left eyes (OS) were 376 ± 20.2 and 370 ± 16.8 μm, respectively. There was positive correlation between the thinnest point and area a in both the right eyes (r = 0.892, P < 0.001) and the left eyes (r = 0.832, P < 0.001). Conclusions  This is the first documentation of the rabbit corneal thickness in vivo using the spectral‐domain AS‐OCT. Pigmented rabbit corneas were almost 150 μm thinner than human corneal values. Gender and eye were not associated with any statistical differences in central corneal thickness in this study.     

Presence of opioid growth factor and its receptor in the normal dog,cat and horse cornea

Objective To determine if opioid growth factor (OGF, [Met⁵]enkephalin) and its specific receptor (OGFr) are present in normal cat, dog and horse cornea. Animals studied Normal dog, cat and horse. Procedure Corneas were obtained from animals euthanized for reasons unrelated to this project. One cornea from each of three normal cats, dogs and horses was evaluated. The right or left cornea from each animal was chosen randomly. Corneas were harvested and placed in corneal storage media for transport to The M.S. Hershey Medical Center of The Pennsylvania State University where immunocytochemistry techniques were used to demonstrate the presence and location of OGF and OGFr. Tissues were rinsed in Sorenson's phosphate buffer, immersed in 20% sucrose in buffer and then snap frozen in isopentane. Corneas were then embedded in OCT medium and 15 µm cryostat sections were created. Presence of OGF was determined by using a polyclonal antibody to [Met⁵]enkephalin and assessing immunoreactivity. OGFr presence was determined by using a previously characterized rabbit polyclonal antibody to the receptor. Results OGF and OGFr were identified in large quantities in the corneal epithelium of all three species. Conclusion Opioid growth factor and its specific receptor are present in the corneal epithelium of normal cats, dogs and horses. OGF is present in the cornea of many species and its presence is theorized to inhibit healing of injured tissue.     

Ophthalmic examination of the captive western lowland gorilla (Gorilla gorilla gorilla).

This study examined the captive western lowland gorilla (Gorilla gorilla gorilla) eye as compared and contrasted with the human eye. Bilateral ophthalmic examinations of western lowland gorillas (n = 5) while under general anesthesia were performed opportunistically, including slit-lamp biomicroscopy, dilated fundus examination, cycloplegic retinoscopy, Schiotz tonometry, corneal diameter and thickness measurements, A-scan and B-scan ultrasonography, keratometry, and cultures of the eyelid margins and bulbar conjunctiva. Mean spherical equivalent refractive error by cycloplegic retinoscopy was +1.20 +/- 0.59 diopters. Mean intraocular pressure by Schiotz tonometry was 12.0 +/- 4.3 mm Hg. Mean optic nerve head cup to disc ratio was 0.42 +/- 0.11. Mean horizontal corneal diameter was 13.4 +/- 0.8 mm, and mean vertical cornea diameter was 12.7 +/- 0.8 mm. Mean central corneal thickness by ultrasound pachymetry was 489 +/- 52 microm. Mean axial length of the eye by A-scan was 22.75 +/- 0.71 mm. Mean lens thickness by A-scan was 4.23 +/- 0.34 mm. Mean anterior chamber depth by A-scan was 4.00 +/- 0.26 mm. Mean keratometry reading was 44.38 +/- 1.64 diopters. Eyelid margin and bulbar conjunctival cultures isolated Candida sp. (n = 5), Staphylococcus aureus (n = 4), Staphylococcus epidermidis (n = 3), Staphylococcus saccharolyticus (n = 3), and Micrococcus sp. (n = 3). This study suggests important similarities between western lowland gorilla and human eyes. These similarities may allow diagnostics, techniques, and equipment for human eye surgery, such as those used for cataract extraction and intraocular lens implantation, to be successfully utilized for gorillas.     

Enhanced expression of cyclooxygenase-2 in glaucomatous dog eyes

Objective Cyclooxygenase‐2 (COX‐2)‐derived prostaglandins (PGs) are shown to play important pathophysiologic roles in various disease states. Recently, the effectiveness of topical PGs in reducing intraocular pressure (IOP) has stimulated further interest in the physiologic function of COX‐2 and PGs in normal and glaucomatous eyes. Therefore, we investigated the cell‐type distribution and expression of COX‐2 in normal and glaucomatous dog eyes. Procedures Using isoform‐specific antibodies, we immunohistochemically evaluated COX‐2 expression in formalin‐fixed and paraffin‐embedded normal (n = 5) and glaucomatous (n = 17) dog eyes. Results In the normal eyes, only minimal COX‐2 immunoreactivity was observed in the ciliary epithelium. In the glaucomatous eyes, COX‐2 expression was further observed in the cornea and corneoscleral limbus. In the cornea, moderate to strong COX‐2 expression was observed in all corneal layers (epithelium, stromal cells and endothelium), with the greatest expression present in the epithelial layer. In the corneoscleral limbus area, COX‐2 immunoreactivity was noted in the stromal cells of sclera, trabecular meshwork and endothelial cells of the angular aqueous plexus. Conclusions Increased expression of COX‐2 in dog glaucomatous eyes suggests that COX‐2‐derived PGs may have a potential role in the pathogenesis of canine glaucoma.     

Use of thermokeratoplasty for treatment of ulcerative keratitis and bullous keratopathy secondary to corneal endothelial disease in dogs: 13 cases (1994-2001)

OBJECTIVE: To evaluate the outcome of thermokeratoplasty for treatment of ulcerative keratitis and bullous keratopathy secondary to corneal endothelial disease in dogs. DESIGN: Retrospective study. ANIMALS: 13 dogs. PROCEDURES: Medical records from 1994 to 2001 for dogs evaluated because of ulcerative keratitis and bullous keratopathy and treated with thermokeratoplasty were reviewed. RESULTS: There were 7 spayed females, 5 castrated males, and 1 sexually intact male, ranging from 6 to 16 years of age. Ten dogs had endothelial degeneration, and 3 dogs had breed-related endothelial dystrophy. All dogs had bullous keratopathy, characterized by microbullae formation that was detected via biomicroscopy. Recurrent or nonhealing corneal ulcers were detected unilaterally in 5 dogs and bilaterally in 8 dogs. Mean +/- SD duration from thermokeratoplasty until ulcerations were healed for all dogs was 2.2 +/- 1.1 weeks. All dogs that underwent thermokeratoplasty for nonhealing corneal ulceration secondary to endothelial disease and corneal edema had epithelial wound healing and resolution of corneal ulceration. Mean duration of treatment (ie, topical treatment required until resolution of ulceration) was significantly less after thermokeratoplasty than duration of treatment (with multiple treatments) prior to referral. CONCLUSIONS AND CLINICAL RELEVANCE: It may be necessary to perform thermokeratoplasty of the entire cornea to prevent recurrence of ulcerations in areas that have not been treated with thermokeratoplasty.     

Keratoconus in a cynomolgus monkey

In a seven-year-old male cynomolgus monkey, erythema of the upper eyelid and forehead and corneal opacity, edema and conical protrusion in the eye were observed. At necropsy, ophthalmological and serological examinations revealed binocular corneal opacity and conical protrusion and a high IgE level, respectively. Thinning of the epithelium and stroma of the cornea were noted histopathologically. At the center of the corneal epithelium, the number of epithelial cells was reduced, their cytoplasm was poorer and the basal cells were flatter than at the periphery. Bowman's membrane was folded with partial loss or breakage. Collagen fibers were compacted or disarranged, and the keratocytes were increased in the stroma, with focal pyknosis or loss of the endothelium and folding of Descemet's membrane. Electron microscopical examination revealed atrophy of the corneal epithelial basal cells. This is the first report of a case of keratoconus in a cynomolgus monkey.     

Rose bengal positive epithelial microerosions as a manifestation of equine keratomycosis

Purpose To describe the clinical appearance of corneal epithelial cell microerosions associated with keratomycosis in the horse. METHODS: Retrospective clinical study. RESULTS: Multifocal, punctate, superficial corneal opacities with positive rose bengal retention were noted in six horses with presumed 'viral keratitis'. Faint fluorescein staining was also present in three cases. Equine herpesvirus tissue culture inoculation was negative for a cytopathic effect in three cases. Aspergillus (n = 3), Curvularia (n = 1), and an unidentified fungus (n = 1) were cultured in five horses, and hyphae found on corneal cytology from the sixth. Mixed bacterial infections were present in three eyes. The eyes of two horses with Aspergillus progressed to deep melting corneal ulcers that required surgical therapy. The microerosions remained superficial, but persistent in the other four eyes. Natamycin was utilized topically in all six horses. Transmission electron microscopy from case 6 revealed mucin layer disruption, an intact corneal epithelial cell layer, and fungal attachment to degenerating epithelial cells. The visual outcome was positive in all six horses, although healing was prolonged (48.5 +/- 14.5 days on average in the horses with no surgery; 62 days on average in the two horses that required surgery). CONCLUSIONS: Complete removal or full-thickness penetration of the corneal epithelial cell barrier may not be necessary to allow fungal adherence and initiation of keratomycosis in the horse. Prior to colonization and invasion of the horse cornea, fungi may induce changes in the mucin layer of the tear film that result in or are associated with rose bengal positive microerosions of the superficial corneal epithelium. Horses with painful eyes, and eyes with superficial, multifocal corneal opacities should have their corneas stained with both fluorescein and rose bengal as fungal microerosions may stain weakly, or not at all, with fluorescein, and may thus be mistaken for presumed 'viral keratitis' of the horse.     

Amniotic membrane transplantation for corneal surface reconstruction after excision of corneolimbal squamous cell carcinomas in nine horses

OBJECTIVE: The purpose of this study was to evaluate the usefulness and effectiveness of permanent amniotic membrane transplantation as an adjunctive treatment to superficial keratectomy alone or combined with strontium-90 irradiation for treatment of equine corneolimbal squamous cell carcinoma (SCC) to decrease corneal scarring and recurrence rate. STUDY: The retrospective case study included 11 horses (n = 12 eyes) diagnosed and treated for ocular SCC that involved the limbus and cornea. Nine of those horses (n = 9 eyes) were treated between 2002 and 2006, with superficial lamellar keratectomy alone or combined with strontium-90 irradiation and followed by placement of a permanent amniotic membrane graft in the surgical defect. The level of scarring (i.e. the clarity of the cornea) resulting with the use of amniotic membrane was subjectively compared to cases where a permanent bulbar conjunctival graft was performed following keratectomy combined with strontium-90 irradiation or cryotherapy (n = 3 eyes). Recurrence was defined as the postoperative and postirradiation regrowth of SCC in the same site and globe. RESULTS: The nine horses that received an amniotic membrane graft after keratectomy alone or combined with irradiation showed a minimal level of scarring in a cornea that regained a greater transparency in comparison to the horses that were treated with a bulbar conjunctival graft. All of the horses that received an amniotic membrane graft had 226 +/- 218 days of follow-up without tumor recurrence (mean +/- SD), ranging from 21 days to 778 days. CONCLUSIONS: The combination of superficial keratectomy alone or associated with beta-irradiation and permanent amniotic membrane transplantation is an effective treatment of corneal or corneolimbal SCC in horses. The placement of an amniotic membrane material represents an alternative surgical procedure to bulbar conjunctival grafts, especially if there is a lack of bulbar conjunctiva tissue available after tumor resection or if a particularly large corneal resection is necessary. The amniotic membrane is incorporated into the corneal defect and seems to create noticeably much less scarring than a corneal defect covered by bulbar conjunctiva.     

Lamellar keratoplasty with a graft of lyophilized acellular porcine corneal stroma in the rabbit

Objective To evaluate the efficacy of lamellar keratoplasty in the rabbit using a graft of lyophilized acellular porcine corneal stroma (APCS). Animal studied Twelve adult 2–2.5 kg Zealand white rabbits were studied. Procedure The cell components of the porcine cornea were removed by the means of enzymatic digestion, freezing, and thawing and then APCS was lyophilized. The 6.5 mm diameter APCS was implanted on a 6.0‐mm diameter keratectomy wound each of 12 rabbits. The postoperative clinical and histological evaluations were performed in the early, intermediate, and late periods. Results All corneal wounds healed. Ten of the 12 grafts of APCS were integrated completely with the receptive cornea except two grafts scraped partially off by the eyelid. The blepharospasm, ocular discharge, and edema of the cornea were marked 1 week after transplantation. New vessels invaded the graft after week 2 and regressed after week 8. The cornea became transparent gradually. The histological evaluation showed that the epithelium on the graft stratified normally post surgery. The keratocytes of the recipient grew into the graft and were proliferative at week 4. The inflammatory cells and new vessels were observed before week 8. The fibrosis in the graft was revealed at week 4 and lessened at week 8. The histological structure of the cornea after surgery was similar to the normal cornea at week 32. Conclusions APCS can recover the integrity of the rabbit's cornea and become transparent in vivo. APCS is an effective graft for lamellar keratoplasty in the rabbit.     

Scanning electron microscopy of bovine corneas irradiated with sun lamps and challenge exposed with Moraxella bovis

Effects of UV radiation and irradiation followed by challenge exposure with Moraxella bovis on the corneal epithelium were studied in 6 calves by scanning electron microscopy. After UV irradiation, the number of dark cells comprising the surface epithelium increased. Many epithelial cells were in various states of degeneration and were characterized initially by large round nuclei, whereas sloughing and peeling were characteristic of the last degenerative stage. All M bovis-infected irradiated eyes had large numbers of degenerating cells, deep epithelial defects, fibrin strands, surface inflammatory cells, and debris. A few M bovis organisms were randomly attached to the cornea before visible ulceration. There were many inflammatory cells between the ulcerated corneal epithelium and adjacent nonulcerated epithelium. Epithelial cells at the margin of the ulcer appeared swollen. Light, dark, and intermediate epithelial cell types could not be distinguished peripheral to the ulcer.     

Density of corneal endothelial cells and corneal thickness in eyes of euthanatized horses

OBJECTIVE: To determine density of corneal endothelial cells and corneal thickness in eyes of euthanatized horses. SAMPLE POPULATION: 52 normal eyes from 26 horses. PROCEDURE: Eyes were enucleated after horses were euthanatized. Eyes were examined to determine that they did not have visible ocular defects. Noncontact specular microscopy was used to determine density of corneal endothelial cells. Corneal thickness was measured, using ultrasonic pachymetry or specular microscopy. RESULTS: Mean density of corneal endothelial cells was 3,155 cells/mm2. Cell density decreased with age, but sex did not affect cell density. Values did not differ significantly between right and left eyes from the same horse. Cell density of the ventral quadrant was significantly less than cell density of the medial and temporal quadrants. Mean corneal thickness was 893 microm. Sex or age did not affect corneal thickness. Dorsal and ventral quadrants were significantly thicker than the medial and temporal quadrants and central portion of the cornea. We did not detect a correlation between corneal thickness and density of endothelial cells in normal eyes of horses. CONCLUSIONS AND CLINICAL RELEVANCE: Density of corneal endothelial cells decreases with age, but corneal thickness is not affected by age or sex in normal eyes of horses. The technique described here may be useful for determining density of endothelial cells in the cornea of enucleated eyes. This is clinically relevant for analyzing corneal donor tissue prior to harvest and use for corneal transplantation.