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.     

Carbon dioxide laser photoablation adjunctive therapy following superficial lamellar keratectomy and bulbar conjunctivectomy for the treatment of corneolimbal squamous cell carcinoma in horses: a review of 24 cases

Objective To determine the complications and nonrecurrence rates following superficial lamellar keratectomy, bulbar conjunctivectomy, and adjunctive carbon dioxide (CO₂) photoablation for corneolimbal squamous cell carcinoma (SCC) in the horse. Study design Retrospective study. Sample population Twenty‐four horses with corneolimbal SCC. Procedure Medical records of horses diagnosed with corneolimbal SCC that was surgically excised and where CO₂ photoablation was used as an adjunctive therapy from 2000 to 2007 were reviewed. Signalment, prior therapy, tumor location and size, complications, and recurrence of SCC were recorded. Results The Thoroughbred was the most commonly (25%) represented breed. Lesions were >10 mm in diameter in 70.8% of cases. Eight horses (33.3%) had neoplastic cells extending to the deep margin of the keratectomy. All horses were available for follow‐up for an average ± standard deviation of 40.7 ± 25 months. Four horses (16.7%) developed a recurrence of SCC. Three of these four horses underwent repeat keratectomy and CO₂ photoablation, one each, at 4 months, 1, and 2 years following the initial procedure. One horse underwent enucleation 8 months following the initial procedure. Conclusions and clinical relevance As an adjunctive therapy, CO₂ photoablation was successful in 87.5% of the horses following a single procedure and in a total of 91.7% following a second therapeutic application. CO₂ photoablation appears to be effective as an adjunctive therapy following removal of large corneolimbal SCC in the horse and in cases in which all tumor cells were not excised.     

Superficial keratectomy and cryosurgery as therapy for limbal neoplasms in 13 horses

Objective To determine the usefulness and clinical outcome of a combined procedure of superficial keratectomy and cryosurgery as a treatment for limbal neoplasms in horses. Study design Retrospective study. Animals Thirteen horses with 14 limbal tumors. Methods Medical records of all patients with limbal tumors, referred to the Department of Equine Sciences of Utrecht University between 1995 and 2002, were retrieved. Patient data were analyzed with respect to signalment, tumor surface area and histologic diagnosis. Surgery, performed under general anesthesia, included surgical debulking of the tumor followed by cryosurgery. Long‐term follow‐up information was obtained by re‐evaluation in the clinic, by the referring veterinarian, and/or by telephone enquiry. Results Therapy was successful in 9 out of 14 eyes after the first attempt (64%); one eye needed retreatment (7%), and four eyes were eventually enucleated (29%). The mean follow‐up period was 4.8 years. The initial surface area of the tumor significantly influenced outcome (P < 0.01). Squamous cell carcinoma was the most predominant tumor type (79%). Haflinger horses accounted for 69% of the cases whereas their occurrence in the overall hospital population is approximately 5%. Conclusions and clinical relevance The described technique of superficial keratectomy and cryosurgery is a simple procedure for the treatment of limbal tumors in equine patients that does not require sophisticated equipment. Nor is it tampered by legal restriction, and appears to be effective in tumors with a small surface area (< 2 cm²). Haflinger horses seem to be predisposed for the development of ocular squamous cell carcinoma.     

A retrospective comparison of surgical removal and subsequent CO2 laser ablation versus topical administration of mitomycin C as therapy for equine corneolimbal squamous cell carcinoma

Objective To compare the complications and nonrecurrence rate following topical mitomycin C (MMC) therapy vs. CO₂ laser ablation for treating equine corneolimbal squamous cell carcinoma (SCC). Study design Retrospective study. Sample population Twenty‐five horses with corneolimbal SCC. Procedures Medical records of horses undergoing surgical tumor resection followed by either topical MMC therapy (0.04%) or CO₂ laser ablation between the years of 2004 and 2010 were reviewed. Recurrence and complications were compared between groups and within MMC subgroups defined by the time at which treatment was initiated relative to surgery. Results Therapy with topical MMC resulted in a nonrecurrence rate comparable to that achieved with CO₂ laser ablation (82.4% vs. 85.7%, respectively). Initiation of MMC following epithelialization of the surgical site a mean of 15 days postoperatively did not result in increased recurrence rates relative to treatment in the immediate postoperative period. Vision‐ or globe‐threatening complications tended to occur with greater frequency in horses receiving topical MMC in the immediate postoperative period (5 of 6 major complications) relative to following epithelialization of the surgical site (1 of 6 major complications). Conclusions Horses receiving adjunctive topical MMC therapy were no more likely to experience tumor recurrence than were horses undergoing CO₂ laser ablation in the horses in this study. Initiation of two to three rounds of MMC following epithelialization of the surgical site results in fewer major complications and achieves comparable disease resolution relative to treatment in the immediate postoperative period.     

p53 protein expression in conjunctival squamous cell carcinomas of domestic animals

The expression of p53 protein was investigated in eight formalin-fixed, paraffin-embedded conjunctival squamous cell carcinomas of five horses and one cow, dog and cat each by an immunohistochemical procedure in order to evaluate protein overexpression. Anti-human p53 protein mouse monoclonal antibodies known to be cross-reactive with p53 protein of the animal species examined were used. Positive p53 nuclear immunostaining was detected in five equine, one bovine and one feline cases. Conversely, no p53 immunostaining was found in the only canine case examined. These results demonstrate a frequent p53 overexpression in conjunctival squamous cell carcinoma that could be related to UV-induced mutations of the p53 tumor suppressor gene.     

Expression of epidermal growth factor receptor (EGFR) and Ki67 in feline oral squamous cell carcinomas (FOSCC)

The aims of this study were to establish expression of epidermal growth factor receptor (EGFR) and Ki67 in 67 archived biopsy samples of feline oral squamous cell carcinomas (FOSCCs) and to establish if the expression of either markers was predictive of survival. Samples were immunohistochemically labelled for the two proteins and scored. Statistical analyses of data, including Kaplan-Meier survival curves, were performed. All samples expressed both markers although levels differed between samples. Median overall survival was 46 days and 1-year survival was 5%. There was no correlation between Ki67 and EGFR scores (Pearson's correlation coefficient, P = 0.861). Low cellular proliferation (low Ki67 score) was positively correlated with an overall longer survival (Log Rank, P = 0.02) and a trend towards better survival for the high EGFR group was observed (Log Rank, P = 0.076). Ki67 and EGFR immunostaining in FOSCC may be of value as biochemical markers for screening of biopsies from cases of FOSCC.     

Photodynamic therapy for the treatment of periocular squamous cell carcinoma in horses: a pilot study

Objective  Local photodynamic therapy (PDT) is a novel cancer therapy in veterinary ophthalmology. A prospective pilot study seeking to demonstrate proof of principle and safety for the treatment of equine periocular squamous cell carcinoma (PSCC) was therefore conducted. We hypothesized that surgical excision with adjunctive local PDT is an effective and safe treatment for equine PSCC.
Procedures  Nine horses (10 eyes) with PSCC were treated with surgical resection, local infiltration of resulting wound beds with 2-[1-hexyloxyethyl]-2-devinylpyropheophorbide-a (HPPH) and irradiation with 665-nm wavelength diode laser. Regular follow-up ophthalmic examinations were performed.
Results  Surgical resection and PDT yielded disease-free intervals of 25–68 months in our study horses as of January, 2008. These results were obtained following a single treatment in seven horses and two treatments in one horse. In one horse, carcinoma in situ developed 2.5 months after partial surgical excision and PDT, requiring local excision under standing sedation.
Conclusions  Preliminary results suggest that surgical resection and adjunctive local PDT is a safe and effective novel treatment for PSCC in horses. More research is needed before PDT for the treatment of equine PSCC can be adequately compared with other current modalities. Important to future investigations regarding PDT, tumor recurrence rate, length of hospitalization, number of treatment episodes required to effect tumor remission, and total treatment costs should be examined in a controlled manner. Our present results and experiences suggest that this treatment may be useful in the treatment of equine PSCC.     

Penile and preputial squamous cell carcinoma in the horse: a retrospective study of treatment of 77 affected horses

Reasons for performing study: The most common penile and preputial neoplasm in the horse is the squamous cell carcinoma (SCC), but no large surveys of treatment and effects of the grade of the tumour, based on the degree of differentiation, on outcome of affected horses are available. Objectives: Analysis of treatment of male horses affected with SCC of the external genitalia and long‐term results of treatment. Methods: Seventy‐seven cases of SCC were evaluated. Data recorded included treatment, outcome, post operative histopathology and retrospective tumour grading. Results: Treatments included: cryosurgery, excision, partial phallectomy, partial phallectomy and sheath ablation, and en bloc penile and preputial resection with penile retroversion and removal of inguinal lymph nodes. The incidence of recurrence after partial phallectomy was 25.6% (10/39) and following incomplete removal was 17.9% (7/39). The incidence of recurrence after en bloc resection with retroversion was 12.5% (1/8). In horses with confirmed inguinal lymph node metastasis, the incidence of recurrence was 25.0% (1/4). Poorly differentiated SCCs were more likely to metastasise than well differentiated SCCs, and there was a greater chance that the treatment would be unsuccessful. The success of treatment, complete removal and in preventing recurrence of the tumour, of male horses with SCC of the external genitalia was 55.7%. Conclusions: Horses that receive only partial phallectomy for treatment for SCC of the external genitalia have a high incidence of recurrence in contrast to horses that receive an en bloc resection. Tumour grading of SCC can help predict prognosis and guide selection of treatment.     

Partial orbital rim resection, mesh skin expansion, and second intention healing combined with enucleation or exenteration for extensive periocular tumors in horses

Ocular and periorbital sarcoids and squamous cell carcinoma are common in equine practice. Extensive involvement of periorbital tissues often necessitates removal of the globe if the function of the eyelids can not be maintained with tumor removal alone. This report describes a modification of the standard enucleation or exenteration technique for cases in which there is insufficient skin to achieve primary closure following complete surgical excision. The caudal portion of the dorsal orbital rim is protuberant; partial excision with an osteotome facilitates skin closure by decreasing the size of the wound. Mesh expansion of skin via multiple rows or parallel stab incisions can also be used as an adjunct to facilitate closure. Four horses underwent enucleation or exenteration using the orbital rim resection and mesh skin expansion techniques for extensive periocular tumors that were unresponsive to prior treatments. Follow-up intervals ranged from 6 to 42 months and no horses had tumor regrowth.     

Intralesional 5‐fluorouracil (5‐FU) for the treatment of eyelid squamous cell carcinoma in 5 horses

Eyelid squamous cell carcinoma in equine patients often presents a therapeutic challenge to practitioners due to the generally large area affected upon presentation. Surgical excision can be curative if wide enough margins are achieved, but this is not often attainable without enucleation. Other alternatives have been examined including cryotherapy, radiotherapy, brachytherapy, intralesional chemotherapy and photodynamic therapy. Intralesional chemotherapy using cisplatin, mitomycin‐C and bleomycin have been shown to be successful in treating eyelid squamous cell carcinomas but may be prohibitive to some owners due to the cost of therapy. To the authors' knowledge, this is the first case series to illustrate the effectiveness of intralesional 5‐fluoruracil in treating large equine eyelid squamous cell carcinomas. Macroscopic reduction in the size of the affected areas was noted in each case together with regression of clinical signs associated with the mass. This, coupled with the relative inexpensive nature of the procedure, makes this technique an attractive therapy for either primary treatment of eyelid squamous cell carcinoma or as a cytoreductive technique prior to surgical excision.     

Evaluation of feline oral squamous cell carcinomas for p16 protein immunoreactivity and the presence of papillomaviral DNA

Oral squamous cell carcinomas (OSCCs) develop commonly in cats. While the cause of the feline neoplasms is unknown, a quarter of human OSCCs are caused by papillomavirus (PV) infection. As PV DNA has been previously detected in a feline OSCC, it was hypothesised that PV infection could be a significant cause of feline OSCCs. Human OSCCs that are caused by PVs contain increased p16^CDKN2A protein (p16), which can be detected using immunohistochemistry. In cats, increased p16 immunoreactivity has been reported within PV-associated skin lesions. This study evaluated p16 immunoreactivity within 30 feline OSCCs. Additionally, PCR was used to amplify PV DNA from the OSCCs. Increased p16 immunoreactivity was present within 2 OSCCs. However, as PV DNA was not amplified from any OSCC in this study, it cannot be confirmed that the increased p16 was caused by PV infection. Therefore, these results do not support the hypothesis that PVs are a significant cause of OSCCs in cats. Loss of p16 expression is considered an important process in the development of human non-PV-induced OSCCs. In contrast, loss of p16 immunoreactivity was only present in 2 feline OSCCs. This suggests that human and feline OSCCs develop due to different molecular mechanisms.     

Investigation of multiple Felis catus papillomavirus types (-1/-2/-3/-4/-5/-6) DNAs in feline oral squamous cell carcinoma: a multicentric study

Recent evidence suggests a possible association of Felis catus papillomavirus type 2 (FcaPV-2) DNA with feline oral squamous cell carcinoma (FOSCC). In this study, type-specific PCR targeting two genes (L1/E6 or E1/E6) of FcaPV-1/-2/-3/-4/-5/-6 was performed to detect viral DNA in a large amount of FOSCC samples collected in Italy and Austria. FcaPV-1/-2/-3/-4/-5 were detected in 7/113 (6.2%), 7/93 (7.5%), 6/113 (5.3%), 1/113 (0.9%) and 2/113 (1.8%) specimens, respectively, with different prevalences in Italian vs. Austrian samples, whilst FcaPV-6 went undetected. Our results confirms that FcaPV-2 is the most prevalent in FOSCC, followed by FcaPV-1/-3 and suggest that FcaPVs have variable circulation rates in European countries.     

Neoadjuvant chemoradiotherapy and surgery as treatment for oral maxillary squamous cell carcinoma in a dog

A gingival maxillary squamous cell carcinoma was diagnosed in a 12-year-old male Yorkshire Terrier. After a complete diagnostic work-up, including a computed tomography scan, the tumour was staged as T3bN1aM0 and considered non-resectable at presentation. The combination of neoadjuvant megavoltage radiotherapy and neoadjuvant and adjuvant chemotherapy with carboplatin and doxorubicin decreased the size of the tumour, allowing for surgery. The dog was free from local disease for 421 days after which it was euthanased at the owners' request.     

Superficial, nonhealing corneal ulcers in horses: 23 cases (1989-2003)

Objective To characterize superficial, nonhealing corneal ulcers in the horse and to assess the affect of age, breed, sex, inciting cause of the ulcer, and treatment on healing time of these ulcers. Design Retrospective study. Animals Twenty‐three horses with superficial, nonhealing corneal ulcerations. Procedure Medical records from 1989 to 2003 of horses diagnosed with superficial, nonhealing corneal ulcers were reviewed. Signalment, duration of clinical signs, ophthalmic abnormalities, treatment and response to treatment were recorded. Horses were treated, in addition to medical therapy, using debridement, debridement and grid keratotomy, superficial keratectomy, and superficial keratectomy with conjunctival graft placement. Results Mean age ± SD of horses was 13.7 ± 5.8 years. The mean time of presence of the corneal ulceration was 38.9 ± 21.3 days. Mean time to complete re‐epithelialization of the corneal ulcers after treatment was 20 ± 14.7 days. Horses treated with debridement alone, grid keratotomy and superficial keratectomy healed in a mean time of 15.3 ± 14.6 days, 16 ± 12.6 days, and 22.8 ± 6.7 days, respectively. Conclusions and clinical relevance Horses treated with a single debridement at initial evaluation healed in a significantly shorter time period than horses treated with grid keratotomy or superficial keratectomy. The latter two procedures may be beneficial in the treatment of nonhealing ulceration in horses, but the results of this study suggest that these procedures should only be performed following failure of the ulcer to heal after epithelial debridement.