Morphology of bovine corneal endothelial cells obtained  with alizarin red and optical microscopy

Natália Méndez; Mariane Gallicchio Azevedo; Luísa Soares Cargnin; Maiara Poersch Seibel; Alessandra Fernandez da Silva; Maria Eduarda Mattos Franceschini; Rafaela Silva Rocha; João Antonio Tadeu Pigatto

doi:10.1590/1809-6891v25e-77041E

Autores

Natália Méndez Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, Rio Grande do Sul, Brasil https://orcid.org/0009-0002-2530-1582
Mariane Gallicchio Azevedo Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, Rio Grande do Sul, Brasil https://orcid.org/0000-0002-8912-1436
Luísa Soares Cargnin Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, Rio Grande do Sul, Brasil https://orcid.org/0009-0002-6542-0324
Maiara Poersch Seibel Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, Rio Grande do Sul, Brasil https://orcid.org/0000-0002-5295-4979
Alessandra Fernandez da Silva Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, Rio Grande do Sul, Brazil https://orcid.org/0000-0001-8858-5207
Maria Eduarda Mattos Franceschini Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, Rio Grande do Sul, Brasil https://orcid.org/0000-0002-6705-2740
Rafaela Silva Rocha Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, Rio Grande do Sul, Brasil https://orcid.org/0000-0001-8856-8239
João Antonio Tadeu Pigatto Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, Rio Grande do Sul, Brazil https://orcid.org/0000-0002-1541-9133

DOI:

https://doi.org/10.1590/1809-6891v25e-77041E

Resumo

O objetivo deste estudo foi determinar a morfologia das células endoteliais nas diferentes regiões do endotélio da córnea bovina saudável por meio de microscopia óptica. Foram estudados 20 globos oculares de 10 bovinos machos da raça Brangus, com idade de 24 meses. O endotélio da córnea foi corado com o corante vital vermelho de alizarina e, em seguida, examinado no microscópio óptico e fotografado. Trinta células endoteliais de cada região da córnea foram incluídas na análise. A morfologia das células endoteliais foi analisada nas regiões central, superior, inferior, lateral e medial da córnea. As comparações entre as regiões foram realizadas usando medidas repetidas de análise de variância (ANOVA). As diferenças foram consideradas estatisticamente significativas em P < 0,05. As células endoteliais normais eram principalmente hexagonais (83,7%), pentagonais (7,45%) e heptagonais (8,8%), com um número mínimo de células de outras formas presentes. Não foram observadas diferenças estatísticas na morfologia das células endoteliais quando comparadas as diferentes regiões da córnea. Em relação à morfologia das células endoteliais não houve diferenças entre as regiões da córnea estudadas.

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Referências

Collin SP, Collin HB. A comparative study of the corneal endothelium in vertebrates. Clinical and Experimental Optometry. 1998;81(6):245-254. Available from: https://doi.org/10.1111/j.1444-0938.1998.tb06744.x.

Abib FC, Holzchuh R, Schaefer A, Schaefer T, Godois R. The endothelial sample size analysis in corneal specular microscopy clinical examinations. Cornea. 2012;31(5):546-550. Available from: https://doi.org/10.1097/ICO.0b013e3181cc7961.

Ong SH, Ang M, Metha JS. Evolution of therapies for the corneal endothelium: past, present and future approaches. British Journal of Ophthalmology. 2021; 105(4):454-467. Available from: https://doi.org/10.1136/bjophthalmol-2020-316149.

Jirsova K, et al. Various approaches to the microscopic assessment of the cornea, visualization and image analysis of the corneal endothelium. In: Light and Specular Microscopy of the Cornea. Springer. 2017;1:59-71. Available from: https://doi.org/10.1007/978-3-319-48845-5_4.

Albuquerque L, Pigatto JAT, Pacicco LVR. Analysis of the corneal endothelium in eyes of chickens using contact specular microscopy. Semina: Ciências Agrárias. 2015; 36(2):4199-4206. Available from: https://doi.org/10.5433/1679-0359.2015v36n6supl2p4199.

Brambatti G, Albuquerque L, Pigatto JAT, Vargas EDB, Neu-mann CF. Corneal endothelial cell density and morphology in rabbits’ eyes using contact specular microscopy. Ciência Rural. 2017;47(12):1-5.Available from: https://doi.org/10.1590/0103-8478cr20170027.

Bercht BS, Albuquerque L, Araujo ACP, Pigatto JAT. Specular microscopy to determine corneal endotelial cell morsphology and morphometry in chinchillas (Chinchilla lanigera) in vivo. Veterinary Ophthalmology. 2015;18(1):137-142. Avail-able from: https://doi.org/10.1111/vop.12236.

Coyo N, Peña MT, Costa D, Ríos J, Lacerda R, Leiva M. Effects of age and breed on corneal thickness, density, and morphology of corneal endothelial cells in enucleated sheep eyes. Veterinary Ophthalmology. 2016;19(5):367-372. Available from: https://doi.org/10.1111/vop.1230.

Brandão CVS, et al. Thickness and density of corneal endothelial cells in ovines. Archives of Veterinary Science. 2006;11(1):16-18. Available from: https://doi.org/10.5380/avs.v11i1.5624.

Coyo N, Leiva M, Costa D, Rios J, Peña T. Corneal thickness, endothelial cell density, and morphological and morphometric features of corneal endothelial cells in goats. American Journal of Veterinary Research. 2018;79(10):1087-1092. Available from: https://doi.org/10.2460/ajvr.79.10.1087.

Kobashigawa KK, et al. Ophthalmic parameters in adult Shih Tzu dogs. Ciência Rural. 2015;45(7):1280-1285. Available from: https://doi.org/10.1590/0103-8478cr20141214.

Hünning PS; Andrade MCC; Carissimi A; Pigatto JAT. Morphology of endothelial cells from different regions of the cornea of dogs. Ciência Rural. 2018;48(10). Available from: https://doi.org/10.1590/0103-8478cr20180596.

Pigatto JAT, Abib FC, Pereira GT, Barros PSM, Freire CD, Laus JL. Density of corneal endothelial cells in eyes of dogs using specular microscopy. Brazilian Journal of Veterinary Research and Animal Science. 2006;43(4):476-480. Available from: doi: 10.11606/issn.1678-4456.bjvras.2006.26462.

Pigatto JAT, et al. Morphological analysis of the corneal endothelium in eyes of dogs using specular microscopy. Pesquisa Veterinária Brasileira. 2008;28(9):427-430. Available from: https://doi.org/10.1590/s0100-736x2008000900006.

Morita H, Shimomura K. Specular microscopy of the corneal endothelial cells in common marmosets. The Journal of Veterinary Medical Science. 1996;58(3):277-279. Available from: https://doi.org/10.1292/jvms.58.277.

Faganello CS, Silva VRM, Andrade MCC, Carissimi AS, Pigatto JAT. Morphology of endothelial cells from different regions of the equine cornea. Ciência Rural. 2016; 46(12): 2223-2228. Available from: https://doi.org/10.1590/0103-8478cr20160216.

Albuquerque L, Pigatto AM, Pigatto, JAT. Evaluation of equine (Equus cabbalus) corneal endothelium stored in EUSOL-C® preservation médium.Semina: Ciências Agrárias. 2020; 41(6) suplement 2: 3155-3164. Available from: https://doi.org/10.5433/1679-0359.2020v41n6Supl2p3155.

Coyo N, Leiva M, Costa D, Molina R, Nicolás O, Ríos J, et al. Endothelial cell density and characterization of corneal endothelial cells in the Tawny Owl (Strix aluco) using specular microscopy. Veterinary Ophthalmology. 2019;22(2):177-182. Available from: https://doi.org/10.1111/vop.12578.

Pigatto JAT, et al. Corneal endothelium of the Magellanic penguin (Spheniscus magellanicus) by scanning electron microscopy. Journal of Zoo and Wild Life Medicine. 2005;36(4):702-705. Available from: https://doi.org/10.1638/05017.1.

Pigatto JAT, et al. Scanning electron microscopy of the corneal endothelium of ostrich. Ciência Rural. 2009;39(3):926-929. Available from: https://doi.org/10.1590/S0103-84782009005000001.

Pigatto JAT, et al. Morphometric analysis of the corneal endothelium of Yacare caiman (Caiman yacare) using scanning electron microscopy. Veterinary Ophthalmology. 2004;7(3):205-208. Available from: https://doi.org/10.1111/j.1463-5224.2004.04025.

Vargas EVB, Pigatto, AM, Rocha, RS, Franceschini MEM, Pigatto, JAT. Specular microscopy of the different regions of the corneain enucleated swine eyes - ex vivo evaluation. Brazilian Animal Science. 2023; 24, e-75138E. Available from: https://doi.org/10.1590/1809-6891v24e-75138E.

Saad HA, Terry MA, Shamie N, Chen ES, Friend DF, Holiman JD, Stoeger C. An easy and inexpensive method for quantitative analysis of endothelial damage by using vital dying staining and adobe photoshop software. Cornea. 2008;27(7):818-824. Available from: https://doi.org/10.1097/ICO.0b013e3181705ca2.

Pigatto AM, et al. Sheep Corneal Endothelium Morphology - Evaluation with Trypan Blue and Alizarin Red. Acta Scientiae Veterinariae,2022; 50:1873. Available from: doi: https://doi.org/10.22456/1679-9216.123885.

Taylor MJ, Hunt CJ. Dual staining of corneal endothelium with trypan blue and alizarin red S: Importance of PH for the dye – lake reaction. British Journal of Ophthalmology. 1981;65(12):815-819. Available from: https://doi.org/10.1136/bjo.65.12.815.

Bahn CF, et al. Postnatal development of corneal endothelium. Investigative Ophthalmology & Visual Science. 1986;27(1):44-51. Available from: https://pubmed.ncbi.nlm.nih.gov/3941037/.

Ubels JL, Pruis RM, Sybesma JT, Casterton PL. Corneal opacity, hydration and endothelial morphology in the bovine cornea opacity and permeability assay using reduced treatment times. Toxicology In Vitro. 2000 14(4): 379-86. Available from: https://doi.org/10.1016/s0887-2333(00)00029-1.

Azevedo MG, et al. Specular microscopy of the corneal endothelial cells of bovines: an ex vivo study. Open Veterinary Journal. 2024;13(12): 1554-1561. Available from: https://doi.org/10.5455/OVJ.2023.v13.i12.5.

BRASIL, Ministry of Science, Technology, and Innovations/National Council for the Control of Animal Experimentation. Official Gazette of the Union, Normative Resolution No. 55, October 5, 2022. Executive Branch, Brasília, DF; 192(1):10. Published in: 07/10/2022.

Canavagh HD, et al. Specular microscopy, confocal microscopy, and ultrasound biomicroscopy: diagnostic tools of the past quarter century. Cornea. 2000;19(5):712-722. Available from: https://doi.org/10.1097/00003226-200009000-00016.

Kafarnik C, Fritsche J, Reese S. In vivo confocal microscopy in the normal corneas of cats, dogs and birds. Veterinary ophthalmology. 2007;10(4):222-230. Available from: https://doi.org/10.1111/j.1463-5224.2007.00543.x.

Wenzel DA et al. Staining of endothelial cells does not change the result of cell density. Cell and Tissue Banking. 2019;20:327-328. Available from: https://doi.org/10.1007/s10561-019-09759-6.

Albuquerque L, Pigatto JAT, Pacicco LVR. Analysis of the corneal endothelium in eyes of chickens using contact specular microscopy. Semina: Ciências Agrárias. 2015;36(2): 4199-4206. Available from: https://doi.org/10.5433/1679-0359.2015v36n6supl2p4199.

Tuft SJ, Coster DJ. The corneal endothelium. Eye. 1990;4(3):389-424. Available from: https://doi.org/10.1038/eye.1990.53.

Tamayo-Arango LJ, Baraldi-Artoni SM, Laus JL, Vicenti FAM, Pigatto JAT, Abib FC. Ultrastructural morphology and morphometry of the normal corneal endothelium of adult cross-bred pig. Ciência Rural. 2009;39(1):117-122. Available from: https://doi.org/10.1590/s0103-84782009000100018.

Franzen AA, Pigatto JAT, Abib FC, Albuquerque L, Laus JL. Use of specular microscopy to determine corneal endothelial cell morphology and morphometry in enucleated cat eyes. Veterinary Ophthalmology. 2010;13(4):222-226. Available from: https://doi.org/10.1111/j.1463-5224.2010.00787.x.

McCarey BE, Edelhauser HF, Lynn MJ. Review of corneal endothelial specular microscopy for FDA clinical trials of refrac-tive procedures, surgical devices, and new intraocular drugs and solutions. Cornea. 2008;27(1):1-16. Available from: https://doi.org/10.1097/ico.0b013e31815892da.

Bu J, et al. Hyperlipidemia affects tight junctions and pump function in the corneal endothelium. The American Journal of Pathology. 2020; 190(3): 563-576. Available from: https://doi.org/10.1016/j.ajpath.2019.11.008.