Specular microscopy of the different regions of the cornea in enucleated swine eyes: ex vivo evaluation / Microscopia especular das diferentes regiões da córnea de suínos: estudo ex vivo

The objective of this study was to determine the endothelial cell density (ECD) and hexagonality of the cornea in the different regions of healthy swine corneal endothelium using specular microscopy. Twenty-four eyeballs from 12 male, 6-month-old Large White pigs (Sus scrofa domesticus) were studied. Contact specular microscopy was performed in the central, superior, inferior, lateral and medial regions. The corneal parameters analysed in this study were ECD and hexagonality. The ECD in the central region was 1865 cells/mm2; in the upper region, it was 1877 cells/mm2, in the lower region, it was 1854 cells/mm2, in the lateral region, it was 1847 cells/mm2, in the medial region, it was 1831 cells/mm2. Hexagonality in the central region, was 53%; in the upper region, it was 54%, in the lower region, it was 54%, in the lateral region, it was 54%, in the medial region, it was 54%. There was no significant difference regarding to the evaluated parameters in all corneal regions evaluated. No statistically significantly differences were observed in ECD and hexagonality between the left and the right eyes. This study demonstrates that ECD and hexagonality of the central cornea area represent the entire endothelial mosaic.

O objetivo do presente estudo foi avaliar a densidade endotelial e a hexagonalidade das células endoteliais nas diferentes regiões da córnea de suínos utilizando a microscopia especular de contato. Foram estudados 24 bulbos oculares de 12 suínos (Sus scrofa domesticus), machos, com seis meses de idade e da raça Large White. A microscopia especular de contato foi realizada nas regiões central, superior, inferior, lateral e medial da córnea. A densidade endotelial média na região central foi de 1865 células/mm2, na região superior foi de 1877 células/mm2, na região inferior foi de 1854 células/mm2, na região lateral foi de 1847 células/mm2 e na região medial foi de 1831 células/mm2. Na região central, a hexagonalidade foi de 53%, na região superior foi de 54%, na região inferior foi de 54%, na região lateral foi de 54%, na região medial foi de 54%. Não foram observadas diferenças significativas na densidade celular e na hexagonalidade nas diferentes regiões da córnea analisadas. Este estudo demonstrou que a densidade endotelial e a hexagonalidade da área central da córnea representam todo o mosaico endotelial.

Sheep corneal endothelium morphology - evaluation with trypan blue and alizarin red

Background: The endothelium is a layer fundamental to maintaining corneal transparency. In ophthalmology, sheep eyes have been used as a model in research related to corneal transplantation. Different techniques have been used to evaluate the corneal endothelium. Concerning vital dyes, corneal endothelial cell analyses have not yet been studied in ovines. The purpose of the present study was to evaluate the morphology of endothelial cells from different regions of the cornea of sheep after staining with alizarin red and trypan blue using an optical microscope. Materials, Methods & Results: Twenty healthy eyes of 10 male sheep obtained from a licensed commercial slaughterhouse were studied. The study was approved by the Research Committee of the Faculty of Veterinary at UFRGS and followed the ethical standards of the Association for Research in Vision and Ophthalmology (ARVO). Immediately after the slaughter, the eyes were enucleated and underwent eye examination. The corneal endothelium was stained with trypan blue and alizarin red and examined and photographed using an optical microscope. The central, superior, inferior, nasal and temporal areas of the cornea were evaluated for cell morphology. Data were compared by t-tests. Differences were considered statistically significant at P < 0.05. Immediately after staining the corneal endothelium, it was possible to examine with an optical microscope, obtain images and analyse the shape of endothelial cells from all regions of the sheep cornea. Polygonal, uniform and continuous cells were observed in all samples studied. Considering all the corneas analysed, cells with 6 sides (75.11%), 5 sides (12.76%) and 4 sides (12.12%) were found. In the central region of the cornea 75.91% of cells with 6 sides, 12.6% of cells with 5 sides and 11.48% with 7 sides were found. In the superior region of the cornea 76.07% of cells with 6 sides, 13.25% with 5 sides and 10.68% with 7 sides were found. In the lower region were found 74.72% of cells with 6 sides, 13% with 5 sides and 12.27% with 7 sides. In the temporal region, 74.14% were 6-sided cells, 11.42% had 5 sides, and 14.43% had 7 sides. Furthermore, in the nasal region, 74.72% of the cells had 6 sides, 13.54% had 5 sides, and 11.73% had 7 sides. No significant differences were found between cell morphology in all corneal regions evaluated. In addition, no significant difference was found when comparing the right eye with the left eye. Discussion: Different methods are used for the analysis of corneal endothelium. For ex vivo research optical microscopy after endothelial staining is an alternative low-cost technique that allows the analysis of all regions of the cornea. Quantitative analyses must characterise the endothelial parameters of the different species. The analysis of the morphology of corneal endothelium with an optic microscope after staining with alizarin red has been described as an effective, rapid and cost-efficient method, since this dye blends with the borated cells, allowing identification. In the present study, using optical microscopy and coloration with alizarin red it was possible to explore and obtain images of the ovine endothelium of all regions of the cornea. In the current study, the endothelium had a predominance of cells will 6 sides in all regions studied. This study allowed us to obtain images of the endothelium as well as quantitative data on the morphology of the different regions of the sheep cornea. This study demonstrated that morphology did not differ between the central and peripheral regions. The findings of this study represent a further source of reproducible data that should be considered when using sheep cornea as ex vivo model for experimental research.

Modified eye evisceration in a tropical screech owl (Megascops choliba)

Background: An adult owl was presented with an injury to the right eye that rendered it blind in that eye. The left eye was normal. Removal of the right eye was recommended and a modified eye evisceration was performed. No complications were observed during or after surgery. The objective of this paper is to describe the modified eye evisceration technique that was successfully used in a tropical screech owl (Megascops choliba). Case: An adult owl was presented with an injury to the right eye that rendered it blind in that eye. Two previous surgical treatments have been carried out but have not been successful. Using a portable slit-lamp (Kowa SL-15®), both eyes were examined. The left eye was normal. Upon ophthalmic examination of the right eye, the owl demonstrated blepharospasm and large central corneal ulcer. Removal of the right eye was recommended. The bird received midazolam hydrochloride [Dormire® - 1 mg/kg, IM] and ketamine hydrochloride [Ketamina® - 5 mg/kg IM] as pre-anesthetic medications. Subsequently, the bird was anesthetized with isoflurane (Isoforine®) by facemask for induction, and then maintained with isoflurane vaporized in 100% oxygen through an endotracheal tube. With the aid of a surgical microscope and microsurgery materials, a modified eye evisceration was performed. Post-operatively, the owl received meloxicam [Maxicam® - 0.5 mg/kg, IM] and tramadol hydrochloride [Cronidor® - 15 mg/kg, orally for 4 days]. The day after surgery, the owl was comfortable and its usual appetite was regained. The patient remained hospitalized for 3 weeks and was evaluated daily. The skin sutures were removed 10 days after the surgical procedure and the surgical wound had healed normally. The patient was reintroduced into the wild after 2 months. During the 6 months post-release, the bird was evaluated once a month, and no complications were observed. Discussion: Severe eye trauma and complicated corneal ulcers are common causes of eyeball removal in birds. In birds, there is a high risk of complications during enucleation. The fragility of the orbital bones makes them susceptible to trauma during the surgery. Evisceration involves the removal of the inner contents of the eye while leaving the cornea and the sclera intact. In the current case, evisceration was chosen because the eye was blind, and maintaining a blind eye would be a source of pain and infection. In the modified evisceration technique, the risk of complications is minimal compared to enucleation, mainly because surgical manipulation is minimal. In our case, the total surgery time was 20 min. Another complication reported after enucleation in birds is the possibility of disfiguring the bird because the removal of the globe disturbs the natural head balance. To avoid these complications, the use of an intraocular prosthesis after evisceration in birds has been performed. However, owls have a tubular-shaped globe with scleral ossicles. These factors could hinder or even prevent the accommodation of a cylindrical silicone prosthesis. In the present case, an intraocular prosthesis implant was never considered due to the unavailability of the prosthesis and to avoid the risk of postoperative complications that have been reported from the literature in dogs. In this case, the owl recovered well from anesthesia without complications, and no postoperative hemorrhage was observed. No signs of pain were observed during the postoperative period and the owl had already shown an appetite and fed on the first postoperative day. The previously published reports using the modified evisceration technique also demonstrated an absence of pain signs during the postoperative period.(AU)

Treatment of limbal melanocytoma with full excision and a fresh homologous corneoscleral graft in a dog

Background: Limbal melanoma has been diagnosed in dogs and due to progression may cause vision loss and eyeballremoval. Definitive diagnosis is made through histopathological examination. Therapeutic options include full thicknessresection and repair by homologous corneal tissue, synthetic graft material, and enucleation. In this report, we describe acase of limbal melanocitoma in a dog that has been treated successfully with fresh homologous corneoscleral graft.Case: A 5-year-old female Labrador was referred to the Ophthalmology Veterinary Section of the Federal University ofRio Grande do Sul (UFRGS), Porto Alegre, Brazil, with a history of a pigmented mass located on the left eye. Ophthalmicexamination revealed a pigmented mass located at the left temporal limbus with corneal involvement. Surgical excisionfollowed by reconstruction using fresh homologous corneoscleral was recommended. The patient was premedicated withacepromazine (0.05 mg/kg, IM) and meperidine (20 mg/kg, IM). Anaesthesia was induced with propofol (10 mg/kg, IV)and maintained with isoflurane. Atracurium (0.2 mg/kg, IV) was administered to maintain a central eye position. The massand a free margin were removed by full-thickness corneoscleral resection. A corneoscleral graft was harvested from a dogthat had been euthanised for reasons unrelated to this study and sutured with 9-0 polyglactin 910 using a simple interruptedpattern. The mass was immediately fixed in 10% neutral buffered formalin and submitted for histological sectioning androutine staining. Based on the histopathological analysis it was confirmed limbal melanocytoma. Postoperative treatmentconsisted...

Treatment of limbal melanocytoma with full excision and a fresh homologous corneoscleral graft in a dog

Background: Limbal melanoma has been diagnosed in dogs and due to progression may cause vision loss and eyeballremoval. Definitive diagnosis is made through histopathological examination. Therapeutic options include full thicknessresection and repair by homologous corneal tissue, synthetic graft material, and enucleation. In this report, we describe acase of limbal melanocitoma in a dog that has been treated successfully with fresh homologous corneoscleral graft.Case: A 5-year-old female Labrador was referred to the Ophthalmology Veterinary Section of the Federal University ofRio Grande do Sul (UFRGS), Porto Alegre, Brazil, with a history of a pigmented mass located on the left eye. Ophthalmicexamination revealed a pigmented mass located at the left temporal limbus with corneal involvement. Surgical excisionfollowed by reconstruction using fresh homologous corneoscleral was recommended. The patient was premedicated withacepromazine (0.05 mg/kg, IM) and meperidine (20 mg/kg, IM). Anaesthesia was induced with propofol (10 mg/kg, IV)and maintained with isoflurane. Atracurium (0.2 mg/kg, IV) was administered to maintain a central eye position. The massand a free margin were removed by full-thickness corneoscleral resection. A corneoscleral graft was harvested from a dogthat had been euthanised for reasons unrelated to this study and sutured with 9-0 polyglactin 910 using a simple interruptedpattern. The mass was immediately fixed in 10% neutral buffered formalin and submitted for histological sectioning androutine staining. Based on the histopathological analysis it was confirmed limbal melanocytoma. Postoperative treatmentconsisted...(AU)

Conjunctival melanoma in a horse treated by tumor resection and cryotherapy

Background: Ocular melanoma is very rare compared to cutaneous melanoma in horses. Definitive diagnosis is made through histopathological examination and treatment options include surgical excision associated with cryotherapy, radiation therapy, and chemotherapy. In this report, we describe a case of conjunctival melanoma in a horse that has been treated successfully with surgical excision associated with cryotherapy.Case: A 15-year-old male Percheron male was referred to the Ophthalmology Veterinary Section of the Federal University of Rio Grande do Sul (UFRGS), Porto Alegre, Brazil, with a history of a pigmented mass located on the lower eyelid of the left eye. Ophthalmologic examination revealed ocular discomfort, secretion and a pigmented mass in the left inferior bulbar conjunctiva. The dermatological examination revealed other melanomas in the perineal region. Complete blood count and serum chemistry profile were within normal ranges and prior to surgery the horse was treated with flunixin meglumine (1.1 mg/kg, IV, q 12 h). Sedation was performed with xylazine (0.4 mg/kg, IV) and detomidine hydrochloride (0.01 mg/kg, IV) and then the animal was placed in a retention trunk. The conjunctival mass was resected with a margin of safety. Liquid nitrogen was applied to the tumor site and the adjacent conjunctiva with a copper cryoprobe with one unit of liquid nitrogen. Histopathological examination revealed neoplastic cells containing pigmented melanocytes in the conjunctival submucosa, confirming the diagnosis of conjunctival melanoma. Postoperative treatment was performed with flunixin meglumine (1.1 mg/kg, IV, q 12 h) for 3 days and topical ophthalmic ointment containing neomycin, polymyxin B sulfate and dexamethasone twice daily for one week. Seven days after surgery, the lesion was healed. The patient was followed for 24 months after excision and there was no evidence of recurrence.[...]

Conjunctival melanoma in a horse treated by tumor resection and cryotherapy

Background: Ocular melanoma is very rare compared to cutaneous melanoma in horses. Definitive diagnosis is made through histopathological examination and treatment options include surgical excision associated with cryotherapy, radiation therapy, and chemotherapy. In this report, we describe a case of conjunctival melanoma in a horse that has been treated successfully with surgical excision associated with cryotherapy.Case: A 15-year-old male Percheron male was referred to the Ophthalmology Veterinary Section of the Federal University of Rio Grande do Sul (UFRGS), Porto Alegre, Brazil, with a history of a pigmented mass located on the lower eyelid of the left eye. Ophthalmologic examination revealed ocular discomfort, secretion and a pigmented mass in the left inferior bulbar conjunctiva. The dermatological examination revealed other melanomas in the perineal region. Complete blood count and serum chemistry profile were within normal ranges and prior to surgery the horse was treated with flunixin meglumine (1.1 mg/kg, IV, q 12 h). Sedation was performed with xylazine (0.4 mg/kg, IV) and detomidine hydrochloride (0.01 mg/kg, IV) and then the animal was placed in a retention trunk. The conjunctival mass was resected with a margin of safety. Liquid nitrogen was applied to the tumor site and the adjacent conjunctiva with a copper cryoprobe with one unit of liquid nitrogen. Histopathological examination revealed neoplastic cells containing pigmented melanocytes in the conjunctival submucosa, confirming the diagnosis of conjunctival melanoma. Postoperative treatment was performed with flunixin meglumine (1.1 mg/kg, IV, q 12 h) for 3 days and topical ophthalmic ointment containing neomycin, polymyxin B sulfate and dexamethasone twice daily for one week. Seven days after surgery, the lesion was healed. The patient was followed for 24 months after excision and there was no evidence of recurrence.[...](AU)

Corneal endothelial cell density and morphology in rabbits eyes using contact specular microscopy / Densidade celular e morfologia das células do endotélio da córnea de coelhos obtidas com microscopia especular

The goal of this study was to describe the normal values of corneal endothelial cell density and hexagonality in healthy rabbits eyes using contact specular microscopy. Both eyes of 18 rabbits, males and females, of different ages were evaluated. The eyes were divided into three groups of 12 each according to the age range of the animals: G1 (6 months old), G2 (12 months old), and G3 (48 months old). Parameters studied included mean endothelial cell density and hexagonality. For Group I, the mean cell density was 2307 cells per mm2. For GII, the mean cell density was 1895 cells per mm2. For G III, the mean cell density was 1818 cells per mm2. Cell density decreased significantly with aging (P 0.001) among the three groups. Pleomorphism in the rabbits of GI was 74.33±10.08. In the rabbits of GII it was 71.83±11.38. In the rabbits of GIII it was 64.02±28.80. Significant differences (P 0.001) were evident between GI and GIII and GII and GIII but not between Groups I and II. The results showed a decrease in number of cells and hexagonality with aging. Corneal endothelium of rabbits suffers changes due to advancing age both in terms of cell density and morphology.

Objetivou-se avaliar a densidade celular e a hexagonalidade do endotélio da cَrnea de coelhos saudلveis usando um microscَpio especular de contato. Os dois olhos de 18 coelhos, machos ou fêmeas, de diferentes idades foram avaliados. Os olhos foram separados em três grupos de 12 olhos cada em funçمo da idade: G1 (6 meses de idade), G2 (12 meses de idade) e G3 (48 meses de idade). Os parâmetros estudados incluيram a densidade celular e a hexagonalidade. Para o Grupo I, a densidade celular média foi de 2307 células por mm2. Para GII, a densidade celular média foi de 1895 células por mm2. Para G III, a densidade celular média foi de 1818 células por mm2. A densidade celular diminuiu significativamente com o envelhecimento (P 0,001) entre os três grupos. O pleomorfismo nos coelhos de GI foi de 74,33±10,08. Nos coelhos do GII foi 71,83±11,38. Nos coelhos de GIII foi de 64,02±28,80. Diferenças significativas (P 0,001) foram evidentes entre GI e GIII e GII e GIII, mas nمo entre os Grupos I e II. Os resultados revelaram diminuiçمo da densidade celular e da hexagonalidade com o avanço da idade. O endotélio da cَrnea de coelhos sofre alteraçُes devido ao avanço da idade tanto na densidade celular e quanto na morfologia.

Corneal endothelial cell density and morphology in rabbits eyes using contact specular microscopy / Densidade celular e morfologia das células do endotélio da córnea de coelhos obtidas com microscopia especular

The goal of this study was to describe the normal values of corneal endothelial cell density and hexagonality in healthy rabbits eyes using contact specular microscopy. Both eyes of 18 rabbits, males and females, of different ages were evaluated. The eyes were divided into three groups of 12 each according to the age range of the animals: G1 (6 months old), G2 (12 months old), and G3 (48 months old). Parameters studied included mean endothelial cell density and hexagonality. For Group I, the mean cell density was 2307 cells per mm2. For GII, the mean cell density was 1895 cells per mm2. For G III, the mean cell density was 1818 cells per mm2. Cell density decreased significantly with aging (P 0.001) among the three groups. Pleomorphism in the rabbits of GI was 74.33±10.08. In the rabbits of GII it was 71.83±11.38. In the rabbits of GIII it was 64.02±28.80. Significant differences (P 0.001) were evident between GI and GIII and GII and GIII but not between Groups I and II. The results showed a decrease in number of cells and hexagonality with aging. Corneal endothelium of rabbits suffers changes due to advancing age both in terms of cell density and morphology.(AU)

Objetivou-se avaliar a densidade celular e a hexagonalidade do endotélio da cَrnea de coelhos saudلveis usando um microscَpio especular de contato. Os dois olhos de 18 coelhos, machos ou fêmeas, de diferentes idades foram avaliados. Os olhos foram separados em três grupos de 12 olhos cada em funçمo da idade: G1 (6 meses de idade), G2 (12 meses de idade) e G3 (48 meses de idade). Os parâmetros estudados incluيram a densidade celular e a hexagonalidade. Para o Grupo I, a densidade celular média foi de 2307 células por mm2. Para GII, a densidade celular média foi de 1895 células por mm2. Para G III, a densidade celular média foi de 1818 células por mm2. A densidade celular diminuiu significativamente com o envelhecimento (P 0,001) entre os três grupos. O pleomorfismo nos coelhos de GI foi de 74,33±10,08. Nos coelhos do GII foi 71,83±11,38. Nos coelhos de GIII foi de 64,02±28,80. Diferenças significativas (P 0,001) foram evidentes entre GI e GIII e GII e GIII, mas nمo entre os Grupos I e II. Os resultados revelaram diminuiçمo da densidade celular e da hexagonalidade com o avanço da idade. O endotélio da cَrnea de coelhos sofre alteraçُes devido ao avanço da idade tanto na densidade celular e quanto na morfologia.(AU)