Morphological and autoradiographic studies on the corneal and limbal epithelium of rabbits

The investigation was centered on the morphological features of the conjunctiva-cornea transition (limbus) of the rabbit eye and the proliferative behavior of its epithelium. The eyes were processed for examination with light and electron microscopy, as well as for autoradiography after intravitreal injection of [H-3]thymidine ([H-3]TdR). At the sites of extraocular muscle insertion, the vascularization of the stroma extended to the peripheral cornea, and the limbal epithelium was thin with its basal stratum made up by clear cuboidal cells. In between the muscle insertions, the cuboidal clear cells, as well as the stroma blood vessels; were scarce. At the light microscope level, the basement membrane was distinct in the cornea but not in the limbus or the conjunctiva. Autoradiographs demonstrated that, at the limbus, the basal cells migrated very quickly to the suprabasal region and remained there up to the 28-day interval. Labeled cells were identified in all epithelial layers of the cornea, including the basal one, at 21 and 28 days but not in the limbal basal clear cells. The rate of renewal of conjunctival epithelium was similar to that observed for the transition with scarce clear cells. The high-resolution autoradiographs demonstrated that the basal cuboidal clear limbal cells exhibit a quick renewal and that they are not label-retaining cells. These latter ones were detected all over the corneal epithelium and in the suprabasal layers of the limbus up to 28 days, in physiological conditions, without the need of stimulation by damage to the corneal epithelium.

Transplante de membrana amniótica canina criopreservada para cicatrização de córnea com deficiência de células límbicas em coelhos

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Morphological analysis of the corneal endothelium in eyes of dogs using specular microscopy

Foram estudados 20 olhos de 10 cães sem raça definida, machos e fêmeas com 6 anos de idade. A morfologia das células do endotélio da córnea foi analisada utilizando-se microscópio especular de contato. Foram estudadas as regiões central e periférica da córnea. de cada região da córnea foram realizadas três micrografias. de cada micrografia foram analisadas no mínimo 100 células endoteliais. Foram obtidos os valores do polimegatismo e pleomorfismo. O endotélio corneano de cães caracterizou-se por uma monocamada de células poligonais uniformes em tamanho e forma. A forma predominante das células endoteliais foi hexagonal. O índice de polimegatismo foi 0.22. Este estudo demonstrou que a morfologia das células do endotélio da córnea de cães é semelhante à observada em humanos.

Excision of sclerocorneal limbus in dogs and resulting clinical events: study of an experimental model

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Aspectos clínicos e imunopatológicos da ceratoplastia com membrana amniótica xenógena fresca e conservada em glicerina: estudo experimental em coelhos

Avaliaram-se, por meio da análise clínica, histopatologia e imunoistoquímica, os efeitos da aplicação da membrana amniótica xenógena fresca e conservada em glicerina, sobre os mecanismos imunológicos da superfície ocular. Para tal, utilizaram-se 40 coelhos, distribuídos em dois grupos experimentais, os quais foram avaliados por 21 dias. A avaliação clínica revelou que a membrana amniótica xenógena conservada em glicerina estimulou uma resposta inflamatória aguda maior que a membrana aplicada fresca. A análise histopatológica indicou que ambas se comportaram de forma semelhante a partir da primeira semana de pós-operatório, apresentando as alterações clássicas da resposta inflamatória da córnea, com o predomínio de infiltrado do tipo polimorfonuclear. A análise imunoistoquímica indicou que, ainda aos 21 dias, a resposta imune local é inespecífica, permitindo concluir que a resposta imune específica na córnea é tardia e que a córnea é um sítio privilegiado para aplicação de enxertos com características imunológicas diferentes, visto que não houve o estímulo para o desenvolvimento de uma resposta mais específica nos grupos avaliados durante toda a execução do experimento.

Superficial keratectomy and 360º conjunctival flap for bullous keratopathy in a dog: a case report

Relata-se o caso ocorrido em um cão, da raça Pinscher, com dois anos de idade e histórico de desconforto no olho direito. O olho esquerdo havia sido enucleado por outro profissional, por apresentar os mesmos sinais, cujo tratamento clínico instituído não lograra êxito. O valor do teste da lágrima de Schirmer encontrava-se aumentado e identificou-se diminuição da pressão intraocular à tonometria de aplanação. Observaram-se, à biomicroscopia, edema corneal profuso e ceratocone, e o teste da fluoresceína foi negativo. Gonioscopia e oftalmoscopia não lograram fornecer dados relevantes dadas as condições da córnea. Diagnosticou-se ceratite bolhosa. Optou-se pelo tratamento cirúrgico, que fora realizado em duas etapas: 1- ceratectomia superficial e flap conjuntival de 360º; 2- ceratectomia superficial para devolver transparência à córnea. Transcorridos 30 dias da segunda ceratectomia superficial, o flap de terceira pálpebra foi desfeito. Observou-se conjuntivalização do quadrante nasal superior da córnea, córnea clara no eixo visual e retorno da visão.

Microbiological profile of donor corneas stored for tectonic transplantation purposes in rabbits

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Experimental lamellar keratoplasty in rabbits using microfibrilar cellulose membrane: clinical, morphological and immunohistochemical findings

Avaliaram-se aspectos clínicos, histopatógicos e imunoistoquímicos da córnes de coelhos da raça Nova Zelândia adultos e machos em ceratoplastias lamelares com membrana de celulose microfibrilar. Trinta animais distribuídos em cinco grupos (n=6) foram estudados por até 60 dias de pós-operatório. A avaliação clínica revelou manifestações moderadas de edema, blefaroespasmo e fotofobia ao segundo dia, evoluindo para formas discretas ou ausentes a partir do sétimo dia, período em que se observou, clinicamente, reparo do defeito corneal. A histopatologia revelou uma fina camada de células escamosas, recobrindo a área lesada já aos sete dias, com discreto infiltrado de células polimorfonucleares. Observaram-se vasos no epitélio a partir do 15o dia, com regressão ao 48o dia. A marcação com o anticorpo Ki67 mostrou aumento de células em proliferação aos 15 dias no epitélio e aos 30 dias no estroma. Nesse período, ocorreram remodelamento e adesão epitealial. Considerando a boa integração do implante, admite-se a membrana de celulose como um bom material a ser utilizado em ceratoplastia lamellar.

Topical 1% Nalbuphine on corneal sensivity and epitheilization after experimental lamellar keratectomy in rabbits

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Use of fresh autogenous vaginal tunic in the experimental lamellar keratoplasty in dogs (Canis familiaris, LINNAEUS, 1758)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Glycosaminoglycans in components of the rabbit eye: synthesis and characterization

Purpose. To trace the eye components involved in proteoglycan synthesis and to characterize the sulfated glycosaminoglycans which are associated to these macromolecules.Methods. Sodium [S-35]-sulfate was injected intravitreally and the rabbits were killed at different time intervals after the injection. The glycosaminoglycans of choroid, ciliary body, cornea, iris, lens capsule, retina and sclera were extracted and processed for estimations of their specific activities, and for electrophoresis plus autoradiography with or without previous treatment with specific enzymes. In addition, methacrylate sections of the eyes were analysed by autoradiography.Results. The peak of specific activities of the glycosaminoglycans of all eye components occurred at 2 days after the intravitreal injection of [S-35]-sulfate. The autoradiography of the agarose gels revealed three types of glycosaminoglycans, namely, heparan-, chondroitin- and dermatan sulfate, only in the retina. The other eye components contained heparan sulfate and either chondroitin or dermatan sulfate. Tissue autoradiography together with the biochemical techniques contributed to unravel the origin of the glycosaminoglycans in the eye components.Conclusions. The results of the present investigation have shown that heparan sulfate, contrasting to chondroitin sulfate and dermatan sulfate, is synthesized in all eye components studied and that the glycosaminoglycan composition differs according to the tissue of origin.

Mast cells in the developing avian eye

Mast cells are present in the eye of Gallus domesticus, appearing in the anterior uvea in embryos at stage 39 HH (13th day). In hatching and adult birds, they are present in the sclera, uvea, pectinate Ligament, and conjunctiva. Mast cells are absent in the cornea, retina, and pecten oculi.Maturing mast cells in the anterior eye segment appear as round cells having eccentric nuclei and a few cytoplasmic metachromatic granules, whose fluorescence increases during development. Mature cells are more numerous in late development, and their cytoplasm is rich in metachromatic and intensely fluorescent granules. Ultrastructurally, maturing mast cells display progranules and a few electron dense and homogeneous granules on one side of the cell. Mast cells of adult birds possess homogeneous cytoplasmic granules, some of which display protuberances that penetrate hollows of adjoining granules. Heterogeneous granules exhibiting latticed and mottled patterns are also present. The existence of mast cells in the anterior eye segment indicates that these cells might perform a physiological role during development and in aqueous humor outflow. They might modulate exchanges between blood and aqueous humor through chemical mediators present in their granules. (C) 1996 Wiley-Liss, Inc.

B-mode ultrasonography biometry of the Amazon Parrot (Amazona aestiva) eye

PurposeUltrasonographic evaluation of the eye is a relatively recent addition to routine ophthalmic diagnostics in small animal ophthalmology. Some parameters for ophthalmic biometry have been established. There are few studies in clinical avian ophthalmology that describe ultrasound images of eye in some nocturnal avian species and in other birds that do not belong to the Brazilian fauna, but the psittacine family is not represented. The purpose of this study was to describe the following measurements: the distances between cornea and anterior lens capsule (D1) between the anterior and posterior lens capsule (D2), between posterior lens capsule and optic papilla (D3) and the axial length.MethodSixty four transpalpebral ocular ultrasound examinations were performed on 32 Blue fronted Amazon parrots (Amazona aestiva) with no history of previous ophthalmic disease.Result and DiscussionThe measurements were taken in sagital planes using a 10 MHz linear probe without a standoff pad. The mean values for the left eye were; D1 0.17 +/- 0.03 cm, D2 0.35 +/- 0.02 cm, D3 0.73 +/- 0.04 cm and the axial length 1.26 +/- 0.06 cm. In the right eye D1 0.17 +/- 0.02 cm, D2 0.34 +/- 0.02 cm, D3 0.74 +/- 0.03 cm and the axial length 1.25 +/- 0.05 cm. No significant statistical difference was observed among the birds or between the left and right eye. The description of these parameters will allow the veterinary practitioner to evaluate the structural changes that specific diseases may cause in these animals.

Use of specular microscopy to determine corneal endothelial cell morphology and morphometry in enucleated cat eyes

The purpose of this study was to investigate the effect of age on endothelial morphology and morphometry in cats. The corneal endothelium was studied using a contact specular microscope. A total of 18 cats (Felis catus Linnaeus, 1758) were evaluated in this study. The subjects were divided into three groups of six cats each in function of age: G1 (1 to 3 months old), G2 (5 to 12 months old), and G3 (24 to 40 months old). The examination presented data as endothelial cell density (ECD), average cell area, corneal thickness, polymegathism, and pleomorphism. Results revealed ECD decrease in corneas of normal cats with age, as well as a corresponding increase in endothelial cell area and pleomorphism. The present work suggests that the endothelial parameters evaluated change with advancing age.

Paracoccidioidomycosis: study of six cases with ocular involvement

We present 6 patients with ocular involvement due to paracoccidioidomycosis. All cases were confirmed by the finding of Paracoccidioides brasiliensis in histopathological or direct mycologic examination of material from the lesion in the eyelid or conjunctiva. In two cases the bulbar conjunctiva was also involved, in another the cornea, and still another patient developed endophthalmitis. The presence of this mutilating disease which may lead to blindness should be suspected when chronic blepharitis or palpebral ulcerated papular lesions are detected in patients from endemic areas of paracoccidioidomycosis. This etiology should also be suspected in patients with anterior and posterior uveitis after discarding the most frequent causes of this condition. © 1988 Kluwer Academic Publishers.