Successful treatment of an unusually large corneal epithelial inclusion cyst using equine amniotic membrane in a dog

A 10‐year‐old intact male Yorkshire Terrier was referred for investigation of a large raised and nonpainful corneal lesion oculus dexter. Clinical examination revealed a pale, translucent corneal mass, which occupied half of the corneal surface and measured 11 mm × 11 mm × 13 mm. The mass was removed by superficial keratectomy and equine amniotic membrane (AM) was transplanted into the large corneal defect to cover the wound and provide tectonic support for the remaining cornea. The mass was histologically confirmed as a corneal epithelial inclusion cyst. There was no evidence of recurrence or complication at the surgical site 100 days postoperatively. Corneal epithelial inclusion cysts are uncommon in dogs. Although superficial keratectomy is the recommended treatment for corneal inclusion cyst, the combination of superficial keratectomy and AM transplantation had to be considered as an alternative for repair of large corneal defects. This is the first case report of the combined application of AM and superficial keratectomy to successfully treat a corneal inclusion cyst in a dog.     

A retrospective study of 30 cases of frozen lamellar corneal graft in dogs and cats

Frozen lamellar corneal grafts and nictitating membrane flaps were used in 18 dogs and 12 cats to repair deep corneal defects. In all dogs either melting corneal ulcers or descemetoceles were present. In the 12 cats, nine had either a melting corneal ulcer or descemetocele, two animals had acute bullous keratopathy, and one cat had corneal sequestrum. Initial vascularization with gradual clearing of the graft occurred during the first 45 days postoperatively. At 60 days postoperatively, all eyes were visual. Frequent postoperative complications included: focal dehiscence of the wound ( n  = 9); melting of part of the graft ( n  = 7); and pigmentation of the graft ( n  = 4). The frozen lamellar corneal graft was a very safe technique, and restored the tectonic and the optical function of the cornea. It provided the best results in corneas with nonperforating corneal defects. This technique provides poorer results when the cornea was perforated prior to surgery or during the surgical procedure.     

A keratoprosthesis prototype for the dog

Objective  To describe the technique for implantation of a novel keratoprosthesis (KP) prototype and evaluate its application for the treatment of corneal blindness in dogs.
Animals studied  Seven dogs, all of them being clinically blind before surgery as a result of severe corneal endothelial disease (5/7) or chronic superficial keratitis (2/7) that were unresponsive to prior therapy.
Procedures  A silicone KP was implanted unilaterally, just anterior to Descemet's membrane, after creating a stromal pocket by deep stromal lamellar dissection.
Results  Implantation of the KP was accomplished without complication in six of seven operated dogs. In the remaining case, an intra-operative complication (perforation of Descemet's membrane) was associated with extrusion of the KP 8 weeks postoperatively. All operated eyes regained limited vision after surgery. Three to six months after implantation purulent keratitis occurred in all five eyes with endothelial disease, necessitating surgical removal of the KP 6 months postoperatively in 5/7 eyes.
Conclusions  This KP prototype shows promise as a treatment for certain blinding corneal diseases. However, changes in the design of this KP, allowing improved stromal integration, will be necessary before its clinical application can be approved.     

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.     

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.     

Corneal sensitivity and aqueous tear production in dogs undergoing evisceration with intraocular prosthesis placement

OBJECTIVE: To determine whether there is an association between evisceration with intraocular prosthesis placement via a dorsal scleral approach and decreased corneal sensitivity or aqueous tear production in dogs. DESIGN: Prospective study. PROCEDURE: Twenty-one dogs scheduled to undergo unilateral evisceration with intraocular prosthesis, and with a normal normotensive contralateral eye, based on slit-lamp biomicroscopy, indirect ophthalmoscopy and applanation tonometry, were included in the study. Central corneal sensitivity was measured with a Cochet-Bonnet esthesiometer prior to (week 0) and 2, 7, 14 and 28 weeks following surgery. Other pre- and postoperative data collected were Schirmer's tear test (STT I), Schirmer's tear test with topical anesthesia (STT II), corneal fluorescein retention, and intraocular pressure (IOP; postoperative control eye only). Axial globe length was determined for both eyes. Scleral incision length in the surgery eye was also recorded. RESULTS: Corneal sensitivity was significantly lower in eyes undergoing evisceration with intraocular prosthesis than in control eyes both preoperatively and postoperatively at all time points. There was no significant difference between preoperative and postoperative corneal sensitivity within the two groups. There was no significant difference in STT I or STT II-values between control eyes and surgery eyes preoperatively. There was a significant difference between preoperative and 2-week postoperative STT I in the surgery eye. There was a significant difference between the preoperative and 28-week postoperative STT II in the surgery eye. The 28-week postoperative STT I-values differed significantly between control and surgery eyes. Corneal sensitivity tended to decrease with increased degree of buphthalmos or with increased scleral incision length, although neither relationship was statistically significant. CONCLUSIONS: Eyes that have sustained chronically elevated intraocular pressure had decreased axial corneal sensitivity compared to normal eyes, which persisted even after intraocular pressure was reduced. The presence of buphthalmos appeared to be a factor in globes developing decreased corneal sensitivity. Evisceration via a dorsal scleral incision did not result in a significant further reduction in axial corneal sensitivity. A mild reduction in aqueous tear production occurred over time in globes following evisceration with intraocular prosthesis.     

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.     

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.     

Establishing and functional testing of a canine corneal construct

Purpose  To provide a model to be used for in vitro studies on drug effects in dogs, this study was conducted to establish a protocol for the construction of a three-dimensional corneal construct. Primary canine corneal cells and a rabbit corneal epithelial (RCE) cell line were used in comparison.
Methods  The corneal construct was assembled step by step in membrane inserts of a six-well plate over a total of 5 weeks, including culture at the air–liquid interface to allow a differentiation of the epithelial cells. The constructs were studied histologically.
Single cell cultures of canine corneal cells as well as RCE cells were stimulated with lipopolysaccharides (LPS) and sodium dodecyl sulfate (SDS). Treatment with different concentrations of dexamethasone was used to test its effects on the cellular prostaglandin E₂ (PGE₂) production. The same experiments were repeated with the corneal constructs and the reactions compared. Expression of the glucocorticoid receptor (GR) in the constructs was studied using immunohistochemistry.
Results  A protocol for the construction of a vital corneal construct was established and the morphological similarity to the canine cornea in vivo shown. The GR protein was detected in all three cell types of the constructs. Stimulation with LPS and SDS led only in the corneal constructs to a significantly increased PGE₂ production, which could be reduced by dexamethasone.
Conclusions  The corneal construct is an interesting system to test drug effects on corneal cells. It allows studies on a cornea-like system including all three major cell types.     

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.     

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.     

Keratoprosthesis with retrocorneal fixation: preliminary results in dogs with corneal blindness

Objective To evaluate the use and complications of a penetrating keratoprosthesis implantation in the management of corneal opacification in dogs. Methods A retrospective clinical study describes the indications for the surgical technique utilized and the outcomes of this procedure in 20 eyes of 19 dogs with blindness of corneal origin. A successful surgical outcome was defined as a clear keratoprosthesis optic and improvement or restoration of functional vision over a follow‐up period ranging from at least 8 months to a maximum of 7 years. Results Eyes with total corneal opacification resulting from chronic superficial keratitis (n = 11), keratoconjunctivitis sicca (n = 5), endothelial dystrophy (n = 3) and chemical burn (n = 1) were treated by unilateral (n = 18) or bilateral (n = 1) full‐thickness implantation of a keratoprosthesis. Keratoprostheses were retained in 15 eyes (75%) which regained vision to the date of reporting. Among these eyes, six had uncomplicated postoperative course, five developed retroprosthetic membranes and four developed granulation tissue over the optic of the keratoprostheses. These complications were successfully removed surgically in the nine eyes. The five remaining eyes (25%) developed serious early postoperative complications, for which enucleation had to be performed. Conclusion In keratopathies in which the corneal opacification could not be treated by standard medical or surgical procedures, this keratoprosthesis appears to be promising to restore vision in chronic superficial keratitis and deep corneal dystrophy. It appears to have a poor prognosis in keratoconjunctivitis sicca in brachycephalic dogs. The post operative complications retro‐prosthetic membranes and granulomatous overgrowth could be treated well.     

Central corneal thickness in koi fish: effects of age, sex, body length, and corneal diameter

OBJECTIVE: To establish the central corneal thickness (CCT) of normal koi fish by ultrasonic pachymetry, and its relationship to age, sex, body length and corneal diameter. METHODS: Age, sex and body length of 33 koi fish (17 male and 16 female fish) were recorded. Horizontal and vertical corneal diameters of each eye were obtained using Jameson calipers. Central corneal thickness of all eyes was measured by ultrasonic pachymetry. Intraocular pressure (IOP) by rebound tonometry was obtained for a subgroup of nine koi (18 eyes). RESULTS: Mean central corneal thickness was 325.9 microm. Central corneal thickness of female koi was greater than CCT of male fish (P < 0.01). Central corneal thickness increased with increasing age overall and within both sexes (P < 0.01). Central corneal thickness increased with increasing body length (P < 0.001). For male and female fish, CCT increased with increasing horizontal and vertical corneal diameters (P < 0.01). Mean horizontal corneal diameter (HCD) was 8.05 mm, mean vertical corneal diameter (VCD) was 7.38 mm, and HCD was consistently greater than VCD. Mean IOP of a subgroup of these koi was 4.9 mmHg by rebound tonometry. CONCLUSIONS: Koi CCT increases with increasing age, body length and corneal diameter.     

Penetrating keratoscleroplasty and bimodal grafting for treatment of limbal melanocytoma in a dog

A 6-year-old, male neutered, German Shepherd dog presented for evaluation of a dark limbal mass OS. Based on the signalment and clinical findings, a presumptive diagnosis of limbal melanocytoma was made. Additionally, OU demonstrated corneal changes consistent with chronic superficial keratitis. Full thickness en bloc resection of the limbal mass was followed with reconstruction and grafting using both frozen cornea and bulbar conjunctiva. Nitrous oxide cryotherapy was utilized as an adjunctive modality for residual neoplastic cell destruction. Histopathologic evaluation confirmed the presumptive diagnosis. At 18 months postoperatively, there was no evidence of recurrence of the limbal melanocytoma. The dog was treated long-term with both topical steroid and tacrolimus 0.03% for control of the chronic superficial keratitis.     

Corneal plaque containing levofloxacin in a dog

A 13‐year‐old castrated male Yorkshire terrier developed a corneal ulcer 2 weeks after intracapsular lens extraction (ICLE) in the right eye. The corneal ulcer was treated with levofloxacin eye drops. A plaque with a white luster developed in the central cornea 2 weeks after treatment with levofloxacin eye drops. The corneal plaque was surgically removed under inhalant anesthesia. The corneal plaque displayed antimicrobial activity against Escherichia coli. Furthermore, levofloxacin content in the plaque was confirmed by matrix‐assisted laser desorption/ionization time‐of‐flight/time‐of‐flight (MALDI‐TOF/TOF) mass spectrometry (MS). The corneal ulcer completely resolved 2 weeks after the surgical removal of the corneal lesion and replacement of levofloxacin eye drops with tobramycin eye drops. Although the topical use of levofloxacin is unlikely to lead to corneal chemical deposits due to the high water solubility of the drug compared to other topical fluoroquinolones, this patient developed corneal plaque of the antibiotic drop.     

Evaluation of topical nalbuphine or oral tramadol as analgesics for corneal pain in dogs: a pilot study

Objective To evaluate the effectiveness of topical nalbuphine or oral tramadol in the treatment of corneal pain in dogs. Animals studied Fourteen male Beagle dogs. Procedures Dogs were divided into three treatment groups and sedated with dexmedetomidine (5 μ/kg IV). A 4 mm corneal epithelial wound was created in the right eye (OD) of all dogs. Sedation was reversed with atipamazole IM. All dogs received pre/post ophthalmic examinations. Post operatively, Group NB (n = 5) received topical 1% preservative‐free nalbuphine OD q8 h and an oral placebo PO q8 h. Group TR (n = 5) received tramadol (4 mg/kg) PO q8 h and topical sterile saline OD q8 h. Group CNTRL (n = 4) received topical sterile saline OD q8 h and an oral placebo q8 h. All dogs received topical 0.3% gentamicin OD TID until healed. Dogs were pain scored using a pain scoring system modified from the University of Melbourne pain scale at 0, 1, 2, 4, and 6 h, then every 6 h by observers masked to treatment, until corneal wounds were healed. Treatment failure was recorded if cumulative pain scores were above a minimum threshold of acceptable pain and rescue analgesia of morphine (1.0 mg/kg IM) was administered subsequently. Result Four dogs in Group NB, one dog in Group TR, and two dogs in Group CNTRL required rescue analgesia. There was no significant difference in the incidence of treatment failure between groups (P = 0.184). Mean time to rescue was 9.16 h. All corneal wounds were healed by 84 h. Conclusions The results of this study suggest tramadol rather than nalbuphine should be further investigated for the treatment of corneal pain.     

Basic fibroblast growth factor stimulates epithelial cell growth and epithelial wound healing in canine corneas

Objective  To evaluate the effect of basic fibroblast growth factor (bFGF) on the proliferation of canine corneal epithelial cells and epithelial wound healing.
Animal studied  Canine corneal epithelial cells from the corneas of euthanized dogs and corneal epithelial wounds on one eye from each of 24 dogs.
Procedures  The proliferation of corneal epithelial cells in vitro was measured using the methylthiazolyl-tetrazolium (MTT) assay. A corneal wound on one eye of each dog was made with a corneal trephine (6 mm diameter). Four concentrations of bFGF, 0, 100, 500, and 1000 ng/mL, were applied to the affected eyes of dogs, t.i.d. Fluorescein staining was used to assess closure of the corneal epithelial wound.
Results  The addition of bFGF resulted in a significant increase in epithelial proliferation at 24 h after culture, except 1 ng/mL bFGF. Cells with all bFGF treatments proliferated significantly at 48 and 96 h compared to those in the non-bFGF group. bFGF at a concentration of 10 ng/mL promoted cell proliferation maximally. The wound healing rate in the bFGF-treated groups was greater than that in the control. All corneal wounds in bFGF-treated corneas closed by day 7, whereas two of six corneal wounds in the control showed poor healing. None of the eyes developed corneal clouding or neovascularization during the experiment.
Conclusions  Basic fibroblast growth factor accelerated the proliferation of canine epithelial cells and effectively promoted corneal epithelial wound healing.     

Corneal stromal sequestration in a dog

A case of corneal sequestrum in a 9-year-old Shih Tzu is reported. On the ophthalmic examination a brown-pigmented ulcer with mild edema and corneal vascularization was present. The brownish plaque was facing an inferior palpebral tumor. A superficial keratectomy followed by a grid keratotomy and removal of the palpebral mass were performed. Histological findings revealed an inflammatory cell infiltration underneath the acellular stromal layers. No melanin granules were observed. No vascular infiltration was present within the necrotic stroma. The surgical area healed and no recurrence has been reported by the owners at the time of writing. To the authors' knowledge, this is the first report of a corneal sequestrum in a dog.     

Ultrastructural findings in feline corneal sequestra

OBJECTIVES: (1) To describe the ultrastructural features of corneal sequestra in cats; and (2) to enhance our understanding regarding the pathogenesis of feline corneal sequestration. METHODS: Nine corneal sequestra were harvested via keratectomy from globes of nine cats. The sequestra were routinely fixed then postfixed for high resolution light and transmission electron microscopy (HR-LM and TEM, respectively). The tissues were embedded in Epon/Araldite. Sections of 0.5-microm thickness were cut and stained with 1% toluidine blue in 1% sodium tetraborate solution for HR-LM. Ultrathin sections were collected on copper grids and stained with uranyl acetate and Sato's lead stain for TEM. Ultrathin sections were examined and the images were captured on an Advantage HR CCD camera using a Hitachi 7500 electron microscope operated at 80 kV. Two healthy corneas from two cats were harvested immediately following euthanasia. These corneal tissues (control samples) were processed in the same manner as the corneal sequestra for HR-LM and TEM. A portion of each sequestrum was also submitted for polymerase chain reaction (PCR) testing for infectious agents including feline herpesvirus-1 (FHV-1), Toxoplasma gondii, Chlamydophila felis and Mycoplasma spp. RESULTS: Ultrastructure of healthy corneal tissues revealed basal corneal epithelial cells aligned adjacent to a thin acellular layer similar to Bowman's layer with underlying tightly packed, regularly arranged, collagen fibrils oriented in different planes. Keratocytes were elongated and had long and irregularly shaped nuclei, and cytoplasm contained rough endoplasmic reticulum and abundant membrane-bound vesicles. In contrast, corneal sequestra contained varying amounts of an amorphous, electron-dense substance, continuous with intact basal epithelial basement membranes peripherally, and overlying corneal ulceration and loosely packed collagen fibrils. Remnants of necrotic keratocytes were seen in spaces between disarranged collagen layers. In all samples, occasional keratocytes exhibited morphology indicative of apoptosis including clumping and margination of chromatin, and shrunken cytoplasm. Varying degrees of inflammation were noted on HR-LM and TEM of affected corneas including peri- and intralesional neutrophils, lymphocytes, plasma cells, and macrophages. Corneal sequestra were FHV-1-positive (n = 3), FHV-1- and T. gondii-positive (n = 1), T. gondii-positive (n = 3), or negative for DNA of these infectious agents (n = 2) using PCR. All corneal sequestra were negative for DNA of Chlamydophila felis and Mycoplasma spp. using PCR. CONCLUSIONS: Apoptosis may play a role in the pathogenesis of feline corneal sequestration independent of the presence of DNA of these infectious organisms. Prospective clinical studies are warranted to further understand the significance of T. gondii in relation to feline corneal sequestration.