Morphology of endothelial cells from different regions of the equine cornea

Faganello, Cláudia Skilhan; Silva, Vanessa Ruiz Moura da; Andrade, Maria Cristina Caldart de; Carissimi, Alícia; Pigatto, João Antônio Tadeu

doi:10.1590/0103-8478cr20160216

Cited by 8 publications

(20 citation statements)

References 33 publications

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“…In addition, endothelial cell borders were visualised, allowing the quantification of the number of sides of each endothelial cell. This methodology enabled the evaluation of the effects of drugs and surgical procedures on the endothelium and the analysis of different means of preserving the cornea and the endothelial cell morphology of a healthy cornea [5,20].…”

Section: Discussionmentioning

confidence: 99%

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Morphology of Endothelial Cells from Different Regions of the Swine Cornea

Clerot

Hünning

Bettio

et al. 2019

Acta Scientiae. Vet.

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Background:The corneal endothelium is a monolayer of polygonal cells which constitute the last layer of the cornea. The integrity of this layer is critical to cornea transparency. The characterization of normal corneal endothelial morphology is important not only to clinical evaluation but also to selection of areas of the cornea with better quality to be employed as donor tissue. The aim of the present study was to evaluate the morphology of endothelial cells from different regions of the swine cornea after alizarin red staining using optical microscopy. Materials, Methods & Results:Twenty-four healthy eyes from 12 swine Large White breed, with 14-monthold, males or females obtained from a licensed Brazilian commercial slaughterhouse were studied. Immediately after humane slaughter, the eyes were enucleated and submitted to ophthalmic examination. Eyes with signs of diseases of the anterior segment were excluded. The cornea, with 3 mm of the sclera, was removed and placed on a glass microscope slide with the endothelial side up. Four radial incisions were made in the periphery of the cornea to better accommodate the cornea on the microscope slide. Alizarin red was diluted in isotonic solution (0.2 g/100 mL) and the pH was adjusted to 4.2 with hydrochloric acid. Three drops of alizarin red were placed on the corneal endothelium. After 90 s, the dye was removed from the cornea with balanced saline solution. The corneal endothelium was examined and photographed using an optical microscope. All evaluations were performed by the same investigator. Photomicrographs were taken of central, superior, inferior, nasal and temporal corneal areas. Parameter studied included endothelial cell morphology. For the statistical analysis, was employed the ANOVA variance test (repeated measures). Differences were considered statistically significant at P < 0.05. Normal endothelium cells were mainly hexagonal (83.7%), pentagonal (7.45%) and heptagonal (8.8%), with a minimal number of cells of other shapes present. There were no significant statistical differences in the proportion of the morphology and the different regions of the cornea (P = 0.31). Discussion: Different techniques are available for the analysis of corneal endothelium, including mainly scanning electron microscopy, specular microscopy and optical microscopy. 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 the borated cells, allowing identification. In the present study, using optical microscopy and coloration with alizarin red it was possible to explore and to obtain images of the swine endothelium of all regions of the cornea. The analysis of the cellular morphology or the percentage of hexagonal cells are among the main parameters used to evaluate the health of the corneal endothelium. In this study, the endothelium had the predominance of the hexagonal shape in all regions studied. In swine, there are no studies...

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Section: Discussionmentioning

confidence: 99%

“…Among the main methods used for endothelial analysis are scanning electron microscopy (SEM), specular microscopy and confocal micrscopy [3,6,14,15]. Is possible to analyze and photograph the endothelium of the cornea under an optical microscope after staining with alizarin red dye [5,10,18,19].…”

Section: Introductionmentioning

confidence: 99%

Morphology of Endothelial Cells from Different Regions of the Swine Cornea

Clerot

Hünning

Bettio

et al. 2019

Acta Scientiae. Vet.

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“…In the present study, the utilization of eyes within one hour after enucleation allowed the maintenance of endothelial structure. Corneal endothelial evaluations using eyes immediately after the death of the animals have been used in studies with specular microscopy, SEM, and optical microscopy with excellent results FAGANELLO et al, 2016;PIGATTO et al, 2004PIGATTO et al, , 2005aPIGATTO et al, ,b, 2009RODRIGUES et al, 2006).…”

Section: Discussionmentioning

confidence: 99%

Evaluation of equine corneal endothelium after exposure to 0.5% indocyanine green - in vitro study

Silva

Andrade

Faganello

et al. 2018

SCA

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The purpose of the study was to investigate whether indocyanine green (ICG) dye damages the corneal endothelium of horses. Twenty-four corneas of 12 healthy equines, males or females, of different ages were used in this study. Only eyes with no ocular findings were used. Randomly, one eye was included in the treatment group and one in the control group. The eyes of the treatment group were exposed for 1 minute to dye ICG 0.5%. After that the endothelium of all eyes was stained with trypan blue and alizarin red S and analyzed and photographed under an optical microscope. Areas with damaged endothelial cells were manually measured and quantified using software for morphometric analysis and expressed as a percentage of cell damage. In all eyes examined areas of cell damage were observed in both corneas of the control group and the treatment group. The mean endothelial damage was 0.8 ± 0.37% in the treatment group and 0.97 ± 0.39% in the control. The Qui-square test stated that treatment and control group were not different. The ICG 0.5% did not cause acute damage to equine corneal endothelium. Key words: Cataract. Indocyanine green. Corneal endothelium. Alizarin red. ResumoO objetivo do estudo foi investigar se indocianina verde (ICG) induz dano nas células do endotélio da córnea de equinos. Vinte e quatro córneas de 12 equinos saudáveis, machos ou fêmeas, de diferentes idades foram estudadas. Somente olhos hígidos foram utilizados. Aleatoriamente, um olho foi incluído no grupo controle e outro no grupo tratamento. Os olhos do grupo tratamento foram expostos durante um minuto à indocianina verde a 0,5%. Posteriormente, o endotélio da córnea foi corado com azul de tripano e vermelho de alizarina, analisado e fotografado usando microscópio óptico. As áreas com células endoteliais danificadas foram aferidas e quantificadas utilizando um software para análise morfométrica. Os valores encontrados foram expressos como percentual de perda celular. Em todos os olhos examinados foram observadas áreas de dano celular, tanto no grupo controle quanto no grupo tratamento. A perda celular endotelial média foi de 0,8±0,37% no grupo tratamento e 0,97 ± 0,39% no grupo controle. O teste Qui-quadrado confirmou que os grupos tratamento e controle não diferiram. Foi possível concluir que a ICG 0,5% não causou dano agudo nas células do endotélio da córnea de equinos. Palavras-chave: Catarata. Indocianina verde. Endotélio corneano. Vermelho de alizarina.

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“…Specular microscopy, optical microscopy, and scanning electron microscopy (SEM) are among the most widely used methods for corneal endothelium analysis (MORITA et al, 1994;PIGATTO et al, 2009;ALBUQUERQUE et al, 2015;COYO et al, 2016;FAGANELLO et al, 2016). There is consensus that the preparation of corneas for studies using SEM causes cell retraction by reducing the original cell area and increasing the endothelial cell density (VIRTANEN et al, 1984;FAGANELLO et al, 2016).…”

Section: Discussionmentioning

confidence: 99%

“…There is consensus that the preparation of corneas for studies using SEM causes cell retraction by reducing the original cell area and increasing the endothelial cell density (VIRTANEN et al, 1984;FAGANELLO et al, 2016).…”

Section: Discussionmentioning

confidence: 99%

Corneal endothelial cell density and morphology in rabbits’ eyes using contact specular microscopy

et al. 2017

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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.

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Morphology of endothelial cells from different regions of the equine cornea

Abstract: The objective of this study was to evaluate the morphology of different regions of the equine cornea using optical microscopy. Both

Cited by 8 publications

References 33 publications

Morphology of Endothelial Cells from Different Regions of the Swine Cornea

Morphology of Endothelial Cells from Different Regions of the Swine Cornea

Evaluation of equine corneal endothelium after exposure to 0.5% indocyanine green - in vitro study

Corneal endothelial cell density and morphology in rabbits’ eyes using contact specular microscopy

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