Autogenous lamellar corneal grafting in dogs

Lamellar, autogenous, corneal grafts were performed on 9 clinically normal dogs (1 eye in each of 2 dogs and both eyes in each of 7 dogs) and 7 dogs with corneal disease (1 eye in each dog). Nylon, polyglycolic acid, polyglactin 910, and polydioxanone suture materials were used in 8-0 to 10-0 sizes. The eyes of the normal dogs were examined weekly for up to 16 weeks; corneal endothelium was reestablished in all eyes by 6 weeks after surgery. The eyes of the dogs with corneal disease were reexamined periodically for up to 9 months. Twenty-two of 23 (95.7%) grafts were considered successful in that they were translucent, and the dogs were able to see after surgery. The suture materials that caused the least tissue reaction were nylon and polyglactin 910. This procedure is indicated for chronic descemetoceles and other perforating corneal disease. It has the advantage of rapidly resolving severe corneal disease, with a high degree of success.     

Histologic evaluation of the immediate effects of diamond burr debridement in experimental superficial corneal wounds in dogs

Purpose To evaluate the corneal changes immediately after diamond burr debridement of superficial corneal wounds in dogs. Spontaneous chronic corneal epithelial defects (SCCEDs) are the most common form of canine recurrent corneal ulcers. The diamond burr has been used in the management of corneal lesions in humans since 1983. Recently, it has been successfully used in the treatment of SCCEDs in dogs; however, little has been documented as to its mechanism of action. Methods Five adult female research dogs euthanized for reasons unrelated to the study were included, providing 10 normal eyes. An excimer laser spatula was used for epithelial removal after delineation with an 8 mm punch biopsy trephine. Diamond burr debridement was performed for 30 and 45 s in five eyes each (groups 1 and 2 respectively). The procedure was performed on the ventral half of the experimental defect as well as ventral normal cornea, immediately after euthanasia, and prior to enucleation. Samples were processed routinely for histologic evaluation and stained with periodic acid–Schiff. Results No stromal defects could be identified under light microscopy. In experimental corneal wounds, multi‐focal areas remained covered by the epithelial basement membrane (BM) after diamond burr treatment in both groups (group 1 = 48%±16SD, group 2 = 26%±12SD). Removal of BM on group 2 was significantly higher than group 1 (P < 0.05). Conclusions The diamond burr allows a safe method of debridement and does not create defects beyond the epithelial BM in corneal wounds in normal dogs. Evaluation of the diamond burr debridement in cases of SCCEDs is warranted.     

Chronic superficial keratitis in dogs: detection of cellular hypersensitivity.

Dogs affected with chronic superficial keratitis (CSK) and clinically normal dogs were tested for cellular hypersensitivity, using the leukocyte migration-inhibition (LMI) technique to 3 ocular antigens (Staphylococcus aureus and corneal and iridal proteins). Affected dogs had statistically significant increases in hypersensitivity cellular responses against corneal and iridal antigens. Affected dogs did not differ from clinically normal dogs in their cell response to S aureus.     

Effect of topical administration of 1% morphine sulfate solution on signs of pain and corneal wound healing in dogs

OBJECTIVE: To evaluate the effect of topical application of a 1% morphine sulfate solution (MSS) on signs of pain and wound healing in dogs with corneal ulcers and examine normal corneas immunohistochemically for the presence of mu and delta opioid receptors. ANIMALS: 12 dogs. PROCEDURE: A 7-mm superficial corneal ulcer was surgically created in the right eye (OD) of 10 dogs, after which gentamicin solution and 1% MSS (n = 6) or saline solution (4) was administered topically OD 3 times daily. Blepharospasm, tearing, conjunctival hyperemia, aqueous flare, esthesiometer readings, and pupil size were recorded before and 30 minutes after treatment in all dogs. Ulcer size and days to completion of healing were recorded. Corneas from 4 treated and 3 control dogs were evaluated histologically. Normal canine corneas from 2 dogs not used in the study were evaluated immunohistochemically for the presence of mu and delta opioid receptors. RESULTS: Dogs treated with MSS had significantly less blepharospasm and lower esthesiometer readings than did control dogs. Duration of ulcer healing and findings of histologic evaluation of corneas did not differ between groups. Numerous delta and infrequent mu opioid receptors were identified in the corneal epithelium and anterior stroma of normal corneas. CONCLUSIONS AND CLINICAL RELEVANCE: Topical use of 1% MSS in dogs with corneal ulcers provided analgesia and did not interfere with normal wound healing. Both mu and delta opioid receptors were identified in normal corneas of dogs, although the mu receptors were present only in small numbers.     

Survey of bacterial microorganisms in the conjunctival sac of clinically normal dogs and dogs with ulcerative keratitis in Fortaleza, Ceará, Brazil

OBJECTIVE: The ocular microflora in dogs has not been established in north-east Brazil. Thus, the main aim of this research was to determine the bacterial microorganisms in the conjunctival sac of clinically normal dogs and dogs with ulcerative keratitis in Fortaleza, Ceará, Brazil. ANIMALS STUDIED: This study included 60 healthy dogs, 15 dogs with unilateral corneal ulcer, and three dogs with bilateral corneal ulcers. Procedure Samples were taken by a calibrated platinum loop (1 microL) placed directly onto the conjunctival sac and on sterile blood agar. The clinical specimens were incubated at 37 degrees C in an atmosphere of 5% CO2 for 48 h. RESULTS: Of the 120 samples from healthy dogs, only 47 (39%) had positive culture for bacteria, while all of the specimens from eyes with corneal ulcer were positive for bacterial growth. The group of dogs with corneal ulcer had a higher (P < 0.05) number of colony-forming units (CFU) per plate than the group of healthy animals. Of the 59 isolates from healthy eyes, only nine (15.3%) had more than 50 CFU per plate, while in the group of dogs with corneal ulcer, 23 (62.2%) of the 37 isolates presented more than 50 CFU per plate. In both groups Gram-positive bacteria (86.5%) predominated over Gram-negative (13.5%). Staphylococcus spp. was the most frequently isolated genus and S. intermedius predominated in both groups. CONCLUSION: The results of our study are directly applicable to initiate rational, preventive and therapeutic measures with greater accuracy in dogs with corneal ulcer.     

Tear production in dogs with epiphora and corneal epitheliopathy

Measurement of tear volume by phenol red thread tear test (PRT), Schirmer's tear test (STT-1) and Schirmer's tear test with topical anesthesia (STT-2), and vital staining with a combination of fluorescein and rose Bengal of the cornea were performed in dogs with normal eyes and those with epiphora. The breeds included the Shih-Tzu ( n  = 26), and five other breeds ( n  = 50). The PRT, STT-1 and STT-2 results from the five breeds of normal dogs were 26.9 ± 3.0 mm, 17.4 ± 4.3 mm, and 8.8 ± 3.2 mm, respectively. The PRT, STT-1, and STT-2 results for the Shih-Tzu eyes were (mean ± standard deviation): 28.2 ± 4.3 mm, 19.5 ± 4.1 mm, and 9.2 ± 4.5 mm, respectively. In the five breeds, corneal epitheliopathy, as evidenced by the retention of topical fluorescein and rose Bengal, occurred in 97% of dogs with epiphora and in 55% of the dogs without epiphora. Also in the dogs with corneal epitheliopathy the STT-2-value was low (4.0 ± 2.8 mm) compared with those eyes without corneal epitheliopathy (8.8 ± 3.1 mm). In the Shih-Tzu breed the STT-2 results were not significantly different between those dogs with corneal epitheliopathy and those with normal corneas. In the non-Shih-Tzu breeds the decrease in basic tear secretion, as measured by the STT-2, is associated with the corneal epitheliopathy.     

The use of porcine small intestinal submucosa for the repair of full-thickness corneal defects in dogs, cats and horses

OBJECTIVE: To evaluate the efficacy of using a porcine small intestinal submucosa (SIS) graft covered by a conjunctival flap for the surgical repair of full-thickness corneal wounds in dogs, cats and horses. PROCEDURE: All records dating from August 1999 to February 2003 from Purdue University Veterinary Teaching Hospital of patients that had undergone ophthalmic surgical procedures and received a SIS corneal graft for a full-thickness lesion were reviewed. Fifteen cases were identified including six dogs, two cats and seven horses. Requirements for inclusion in this study were that SIS was used as a corneal graft in a full-thickness corneal defect and that the graft was completely covered with a conjunctival flap. RESULTS: Of the 15 cases, one canine patient had received SIS following removal of an epibulbar melanocytoma. The remaining five canine patients had undergone this surgical procedure for the repair of corneal perforation. The two feline patients had been presented for corneal perforation following chronic ulceration. One equine patient had been presented for a deep melting ulcer, three for stromal corneal abscesses, and three for corneal perforations. Complications encountered postoperatively included aqueous leakage, conjunctival flap dehiscence, synechia, cataract and fibrin in the anterior chamber. Fourteen out of 15 patients were visual at the final re-evaluation. CONCLUSION: SIS is an inexpensive, easy-to-handle biomaterial that appears to be suitable for the repair of full-thickness corneal wounds in dogs, cats and horses. Results of our study support the conclusion that this relatively new product is an effective alternative to traditional implantation materials utilized in veterinary ophthalmology.     

Surgical repair of deep melting ulcers with porcine small intestinal submucosa (SIS) graft in dogs and cats

PURPOSE: To evaluate the efficacy of using a porcine small intestinal submucosa (SIS) graft for the surgical repair of deep melting ulcers in dogs and cats. METHODS: Two cats and five dogs presented with deep and large melting ulcers of the cornea. In each case, the necrotic and collagenolytic tissue of the cornea was removed by keratectomy. A SIS graft, 1 mm greater than the corneal defect, was rehydrated in sterile saline and sutured to the edges of the ulcer with a simple interrupted pattern of 9/0 polyglactin 910. A nictitating membrane flap was utilized in two cats and four dogs for 2 weeks. All cases were treated postoperatively with topical and systemic antibiotics, a systemic anti-inflammatory drug and topical atropine. All animals were re-evaluated 15 days, 4 weeks, 35-45 days, 2-3 months and 6 months postsurgery. RESULTS: At 15 days postsurgery, a superficial intense corneal neovascularization surrounded the SIS graft. No ocular discomfort was present and fluorescein staining was negative in all cases. At 4 weeks the SIS graft was thick and opaque in all cases, although in one cat the SIS graft had partially detached. Between 35 and 45 days, SIS graft integration was evident in all eyes, and corneal neovascularization had decreased progressively. All eyes healed without complications and retained corneal transparency. This occurred even in the presence of corneal perforation in two cases: one prior to and one during surgery. CONCLUSION: Results of our study suggest the SIS graft may be an effective alternative surgical treatment to the traditional conjunctival grafts commonly used to repair melting ulcers in dogs and cats. The advantages of using a SIS graft include good corneal transparency, preservation of corneal integrity and maintenance of vision.     

Corneal anatomical differences in the aetiology of chronic superficial keratitis

The apparently normal corneas of 14 diseased eyes of German shepherd dogs (GSDs) were shown to possess very thin epithelial layers on microscopy. Normal corneas of 38 GSDs and 113 dogs of various breeds and crossbred animals were compared for epithelial thickness, structure of corneal stroma and corneal strength. Due to lower values for these criteria found in the GSDs they are considered to play an important role in the aetiology of chronic superficial keratitis.     

Use of porcine small intestinal submucosa for corneal reconstruction in dogs and cats: 106 cases

Objectives : To describe the efficacy of porcine small intestinal submucosa in corneal reconstructive surgery in dogs and cats through a large retrospective study. Methods : A retrospective evaluation of 106 cases of surgical reconstruction of the cornea with small intestinal submucosa seen between May 2005 and January 2010 was carried out. The corneal defect was filled by microsurgical grafting of porcine small intestinal submucosa. The biomaterial implant was deposited in one or several layers depending on the depth of the defect. The animals were examined 3, 6 and 12 weeks after surgery. Results : Vision was preserved in all eyes at three months post‐surgery. In 74 cases (69.8%) the corneal scar was either transparent or discrete, whilst in 32 cases (30.2%) a mild or marked scar was observed. Minor complications occurred in 9 cases (8.5%) with partial integration of the small intestinal submucosa and in 24 cases (22.6%) with faint or mild corneal pigmentation, without impairing vision. In cases followed over a period longer than three months, major complications occurred in five dogs resulting in vision impairment because of pronounced pigmentation. Clinical Significance : Corneal grafting of porcine small intestinal submucosa is an effective method for corneal reconstruction resulting in corneal transparency in most cases. It is an excellent alternative to conventional conjunctival grafts.     

Corneal squamous cell carcinoma in two dogs

Purpose To report two cases of corneal squamous cell carcinoma (SCC) in dogs. Methods Corneal tumors were resected by superficial keratectomy in two cases. Immunohistochemistry of the corneal tissues was performed using anti‐p53 antibody. Results The prominent features of the cases were a clinical history of pigmentary keratitis and chronic keratitis. In each case, a corneal mass was surgically removed with a superficial keratectomy and histologically diagnosed as corneal SCC. Both masses were negative for p53. To reduce chronic corneal irritation, 0.1% hyaluronate sodium ophthalmic solution was applied. After more than 15 months of postsurgical follow‐up there has been no recurrence of either neoplasm. Conclusion and discussion Chronic corneal irritation was suspected as the primary etiology for the corneal SCC. Appropriate surgical removal of the mass and subsequent conservative treatment for keratitis provided effective therapy in these two cases.     

Calcareous degeneration of the canine cornea

The purpose of this paper is to describe a specific presentation of canine corneal calcification. Fourteen cases are described. In seven cases the corneal lesions were bilaterally symmetrical. In five cases the corneal lesion was unilateral. Two dogs were uniocular, the contralateral eye had been enucleated between 1 and 3 months previously by the referring veterinary surgeon following corneal ulceration and perforation. Of a total of 21 eyes with corneal calcification, 16 eyes had associated ulceration. The ulceration presented as follows: two eyes had descemetocoeles, four eyes had corneal perforations, eight eyes had stromal ulceration, and two eyes had superficial punctate ulceration. The cause of the corneal mineralization remains undetermined but underlying systemic disease, particularly hyperadrenocorticism (Cushing’s Syndrome), is suspected as a possible contributing factor in some of these cases. Histopathology was carried out on three cases following a keratectomy and placement of a conjunctival pedicle flap into the ulcerated lesion.     

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.     

Microstructure and glycosaminoglycan ratio of canine cornea after reconstructive transplantation with glycerin-preserved porcine amniotic membranes

Objective  Although amniotic membranes of canine, feline, and equine species have some advantages as corneal transplantation material in many canine ocular diseases, their softness, thinness, and low availability can pose problems. As an alternative, the more abundant porcine amniotic membranes may be used. This paper describes the use of glycerin-preserved porcine amniotic membranes in corneal transplantation in eight normal dogs.
Method  A 0.4-mm deep recipient bed in the axial cornea of the OS of all dogs was created using an 8-mm Barron radial vacuum trephine. The recipient bed was then filled with amnion, and the entire cornea was covered with another piece of the glycerin-preserved membrane. The ocular signs evaluated were corneal opacity and corneal vascularization. The dogs were euthanized on days 5, 10, 20, or 40 after surgery, and samples were collected to evaluate corneal thickness, parenchymal cell number, mean collagen fibril diameter, collagen fibril content and the glycosaminoglycan (GAG) ratio.
Results  Corneal opacity was observed immediately after surgery. Restoration of corneal transparency, regression of corneal vascularization, and visualization of the pupil and iris were noted on day 40.
Conclusions  The clinical observations were supported histologically by regained corneal thickness, parenchymal cell number, mean collagen fibril diameter, collagen fibril content, and GAG ratio, suggesting that this technique may be a novel method for the treatment of ocular surface disorders.     

Use of a dermal biopsy punch for removal of ectopic cilia in dogs: 19 cases

OBJECTIVE: To evaluate the use of a dermal biopsy punch for accurate and quick removal of ectopic cilia in dogs. ANIMALS STUDIED: This study reports the treatment of ectopic cilia detected in 12 dogs during the period from June 2002 to December 2004. Five cases were affected with ectopic cilia in the right eye, and five in the left eye, with two cases affected bilaterally. Nineteen ectopic cilia were detected in 12 dogs. PROCEDURE: The technique described in this study consists of using a 2-3-mm skin biopsy punch with the aid of an operating microscope. RESULTS: No recurrence was noted in any of the cases. The presurgical blepharospasm due to the presence of the cilia resolved as well as uneventful corneal healing of any associated corneal erosions or ulcers. CONCLUSIONS: This method offers a simple, practical alternative to the traditional scalpel procedure as described in the veterinary ophthalmic literature.     

Corneal endothelial cell morphology of normal dogs in different ages

Endothelial cell function is essential to maintain corneal transparency, but unfortunately the regenerative capacity of the endothelium is limited. There are only a few reports describing the effect of age on morphologic appearance of corneal endothelial cells of dogs. Studies of normal corneal endothelial cells in humans and dogs have shown a decrease in endothelial cell density (ECD) and an increase in pleomorphism and polymegethism with advancing age. The purpose of this study was to investigate the effect of age on ECD and endothelial cell morphology in dogs. A total of 30 dogs were divided into three groups (10 dogs/group) based on age: group 1 (2-12 months old), group 2 (24-72 months old), and group 3 (84 months or older). Corneas were processed for light and scanning electron microscopy. Results showed only difference in cell density between group 1 and groups 2 and 3, showing an initial decrease in cell density as the animal matured. Whereas there was significantly greater variation in cell size within the dogs in group 3 than there was within the other two groups, suggesting that there was increased polymegethism and pleomorphism with advancing age.     

Duration of effect and effect of multiple doses of topical ophthalmic 0.5% proparacaine hydrochloride in clinically normal dogs

OBJECTIVE: To determine the duration of effect and the effect of multiple doses of topical ophthalmic application of 0.5% proparacaine hydrochloride on corneal sensitivity in clinically normal dogs. ANIMALS: 8 clinically normal dogs. PROCEDURE: Dogs were randomly allocated to treatment order in a 2 x 2 (period X treatment) crossover study. Treatments consisted of topical application of ophthalmic 0.5% proparacaine (1 drop or 2 drops at a 1-minute interval); treatments were applied to both eyes. A Cochet-Bonnet aesthesiometer was used to determine corneal touch threshold (CTT) before corneal application, 1 and 5 minutes after corneal application, and at 5-minute intervals thereafter for 90 minutes. RESULTS: The CTT value before treatment differed significantly from CTT values after treatment until 45 minutes after application in the 1-drop group and until 55 minutes after application in the 2-drop group. As determined by use of the Cochet-Bonnet aesthesiometer, a significantly greater anesthetic effect was detected for the 2-drop treatment, compared with the effect for the 1-drop treatment, at 30, 35, 40, 45, 50, and 55 minutes after application. Maximal anesthetic effect lasted for 15 minutes for the 1-drop treatment and 25 minutes for the 2-drop treatment. CONCLUSIONS AND CLINICAL RELEVANCE: Duration of corneal anesthetic effect induced by topical ophthalmic application of 0.5% proparacaine in dogs of this study is considerably longer than that reported elsewhere. Serial application of doses of 0.5% proparacaine increases the duration and magnitude of corneal anesthetic effects.     

Amiodarone-induced keratopathy in healthy dogs

Amiodarone has a broad spectrum as an antiarrhythmic agent and is indicated for patients with atrial and ventricular arrhythmias. Amiodarone-induced corneal deposits are the most common reversible side effect (70-100%) in humans. Additional ocular effects in humans include deposits in the lens, retina and optic nerve. This study was conducted to determine ocular effects of chronic oral amiodarone in healthy dogs. Six chronically amiodarone-treated dogs and four controls were used for this study. Ophthalmic examination was performed using biomicroscopy and indirect ophthalmoscopy at the end of 4th, 7th and 11th weeks when dogs received amiodarone. Corneal microdeposits were observed at the end of the 7th week in one eye and at end of the 11th week in the other eye of one dog. Immediately following euthanasia, corneas and optic nerves were harvested for light and electron microscopic analysis. Light microscopic analysis showed corneal deposits in the basal epithelial cells of the cornea of the clinically affected dog. In addition, a significant increase in basal cell turnover as indicated by mitotic index was observed in the affected dog compared to both nondeposit amiodarone and control groups. All remaining animals were normal. One out of six dogs treated with amiodarone demonstrated corneal deposits (16%). This prevalence is low compared to humans. Explanations for this may include species variations particularly in volume of lacrimal secretion, or the need for longer administration. In addition, sunlight is believed to exacerbate the corneal deposits in humans and all dogs in this study were housed indoors.     

Immune-mediated canine and feline keratitis.

Although the normal cornea is devoid of vasculature and lymphatics, there are still several immune-mediated corneal conditions that can occur in dogs and cats. An overview of corneal immunology is presented. Diseases of dogs, including chronic superficial keratitis, superficial punctate keratitis, and canine adenovirus endotheliitis, as well as feline diseases, including eosinophilic keratitis and herpesvirus-related conditions, are discussed.     

Superficial corneal squamous cell carcinoma occurring in dogs with chronic keratitis

Objective Canine corneal squamous cell carcinoma (SCC) is a rare tumor, with only eight cases previously published in the veterinary literature. The Comparative Ocular Pathology Lab of Wisconsin (COPLOW) has diagnosed 26 spontaneously occurring cases, 23 in the past 4 years. This retrospective study describes age and breed prevalence, concurrent therapy, biologic behavior, tumor size and character, and 6‐month survival rates after diagnosis. Results A search of the COPLOW database identified 26 corneal SCC cases diagnosed from 1978 to 2008. There is a strong breed predilection (77%) in brachycephalic breeds, particularly those prone to keratoconjunctivitis sicca. The mean age was 9.6 years (range 6–14.5 years). Follow‐up information >6 months was available for 15 of 26 cases. Recurrence occurred in the same eye in nine cases, seven of which were incompletely excised at the time of first keratectomy. No cases were known to have tumor growth in the contralateral eye and no cases of distant metastases are known. Where drug history is known, 16 of 21 dogs had a history of treatment with topical immunosuppressive therapy (cyclosporine or tacrolimus) at the time of diagnosis. Conclusion Chronic inflammatory conditions of the cornea and topical immunosuppressive therapy may be risk factors for developing primary corneal SCC in dogs. SCC should be considered in any differential diagnosis of corneal proliferative lesions. Superficial keratectomy with complete excision is recommended, and the metastatic potential appears to be low.