Clinical features and corneal optical anisotropies in a rabbit model of limbal stem cell deficiency

Detalhes bibliográficos
Autor(a) principal: Kobashigawa, Karina Kamachi [UNESP]
Data de Publicação: 2018
Outros Autores: Aldrovani, Marcela [UNESP], Barros Sobrinho, Alexandre A. F. [UNESP], Santo Silva, Paloma do Espirito [UNESP], Marcusso, Paulo F. [UNESP], Marinho-Neto, Fausto A. [UNESP], Martines Padua, Ivan R. [UNESP], Laus, Jose Luiz [UNESP]
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UNESP
Texto Completo: http://dx.doi.org/10.5935/0004-2749.20180076
http://hdl.handle.net/11449/164620
Resumo: Purposes: To investigate the intra-laboratory reproducibility of clinical features and to evaluate corneal optical anisotropies in a rabbit model of limbal stem cell deficiency. Methods: Limbal injury was induced in the right eye of 23 adult New Zealand White rabbits using a highly aggressive protocol that combined 360 degrees limbal peritomy, kerato-limbectomy, alkaline chemical burn, and mechanical removal of the epithelium. Clinical evaluation of the injured eyes was performed for 28 days and included corneal impression cytology. Corneas with a severe clinical outcome set typical of limbal stem cell deficiency were then collected, subjected to a histopathological examination, and examined for optical anisotropies. Corneas from healthy rabbit eyes were used as controls. Differences in optical path due to stromal collagen birefringence, as well as linear dichroism related to the expression and spatial orientation of glycosaminoglycan chains from proteoglycans, were measured from cross-sections under a quantitative polarized light microscope. Results: One eye showed signs of hypopyon and was excluded. Signs of ocular inflammation were observed in all eyes studied (n=22). Corneal impression cytology did not detect goblet cells. Twelve of the 22 corneas presented a clinical outcome set typical of limbal stem cell deficiency, which is characterized by the presence of epithelial defects, inflammatory cells, moderate-to-severe opacity, and neovascularization. Microscopic studies under polarized light revealed that relative to controls, limbal stem cell deficiency caused a 24.4% increase in corneal optical path differences. Further, corneas with limbal stem cell deficiency were less dichroic than controls. Conclusions: These results suggest that rabbit models of limbal stem cell deficiency must be rigorously screened for use in preclinical studies to ensure experimental homogeneity because protocols used to create limbal stem cell deficiency could be not associated with good intra-laboratory reproducibility of clinical features. Limbal stem cell deficiency, as induced herein, altered the optical anisotropic properties of the corneal stroma. Such alterations are indicative of changes in collagen packing and the spatial orientation of glycosaminoglycan chains from proteoglycans. Knowledge of these changes is important to potentiate strategies aimed at restoring the morphofunctional integrity of the corneal stroma affected by limbal stem cell deficiency.
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spelling Clinical features and corneal optical anisotropies in a rabbit model of limbal stem cell deficiencyBirefringenceCorneal stromaAnisotropyStem cellsLimbus corneaeGlycosaminoglycansPurposes: To investigate the intra-laboratory reproducibility of clinical features and to evaluate corneal optical anisotropies in a rabbit model of limbal stem cell deficiency. Methods: Limbal injury was induced in the right eye of 23 adult New Zealand White rabbits using a highly aggressive protocol that combined 360 degrees limbal peritomy, kerato-limbectomy, alkaline chemical burn, and mechanical removal of the epithelium. Clinical evaluation of the injured eyes was performed for 28 days and included corneal impression cytology. Corneas with a severe clinical outcome set typical of limbal stem cell deficiency were then collected, subjected to a histopathological examination, and examined for optical anisotropies. Corneas from healthy rabbit eyes were used as controls. Differences in optical path due to stromal collagen birefringence, as well as linear dichroism related to the expression and spatial orientation of glycosaminoglycan chains from proteoglycans, were measured from cross-sections under a quantitative polarized light microscope. Results: One eye showed signs of hypopyon and was excluded. Signs of ocular inflammation were observed in all eyes studied (n=22). Corneal impression cytology did not detect goblet cells. Twelve of the 22 corneas presented a clinical outcome set typical of limbal stem cell deficiency, which is characterized by the presence of epithelial defects, inflammatory cells, moderate-to-severe opacity, and neovascularization. Microscopic studies under polarized light revealed that relative to controls, limbal stem cell deficiency caused a 24.4% increase in corneal optical path differences. Further, corneas with limbal stem cell deficiency were less dichroic than controls. Conclusions: These results suggest that rabbit models of limbal stem cell deficiency must be rigorously screened for use in preclinical studies to ensure experimental homogeneity because protocols used to create limbal stem cell deficiency could be not associated with good intra-laboratory reproducibility of clinical features. Limbal stem cell deficiency, as induced herein, altered the optical anisotropic properties of the corneal stroma. Such alterations are indicative of changes in collagen packing and the spatial orientation of glycosaminoglycan chains from proteoglycans. Knowledge of these changes is important to potentiate strategies aimed at restoring the morphofunctional integrity of the corneal stroma affected by limbal stem cell deficiency.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Univ Estadual Paulista, Fac Ciencias Agr & Vet, Dept Clin & Cirurgia Vet, Jaboticabal, SP, BrazilUniv Estadual Paulista, Fac Ciencias Agr & Vet, Dept Patol Vet, Jaboticabal, SP, BrazilUniv Estadual Paulista, Fac Ciencias Agr & Vet, Dept Clin & Cirurgia Vet, Jaboticabal, SP, BrazilUniv Estadual Paulista, Fac Ciencias Agr & Vet, Dept Patol Vet, Jaboticabal, SP, BrazilFAPESP: 2012/17308-5FAPESP: 2013/01494-7FAPESP: 2014/18007-4CNPq: 467289/2014-0Consel Brasil OftalmologiaUniversidade Estadual Paulista (Unesp)Kobashigawa, Karina Kamachi [UNESP]Aldrovani, Marcela [UNESP]Barros Sobrinho, Alexandre A. F. [UNESP]Santo Silva, Paloma do Espirito [UNESP]Marcusso, Paulo F. [UNESP]Marinho-Neto, Fausto A. [UNESP]Martines Padua, Ivan R. [UNESP]Laus, Jose Luiz [UNESP]2018-11-26T17:55:20Z2018-11-26T17:55:20Z2018-09-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article384-392application/pdfhttp://dx.doi.org/10.5935/0004-2749.20180076Arquivos Brasileiros De Oftalmologia. Sao Paulo: Consel Brasil Oftalmologia, v. 81, n. 5, p. 384-392, 2018.0004-2749http://hdl.handle.net/11449/16462010.5935/0004-2749.20180076WOS:000443858200006WOS000443858200006.pdfWeb of Sciencereponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengArquivos Brasileiros De Oftalmologia0,518info:eu-repo/semantics/openAccess2024-06-07T13:01:45Zoai:repositorio.unesp.br:11449/164620Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T15:49:22.425965Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv Clinical features and corneal optical anisotropies in a rabbit model of limbal stem cell deficiency
title Clinical features and corneal optical anisotropies in a rabbit model of limbal stem cell deficiency
spellingShingle Clinical features and corneal optical anisotropies in a rabbit model of limbal stem cell deficiency
Kobashigawa, Karina Kamachi [UNESP]
Birefringence
Corneal stroma
Anisotropy
Stem cells
Limbus corneae
Glycosaminoglycans
title_short Clinical features and corneal optical anisotropies in a rabbit model of limbal stem cell deficiency
title_full Clinical features and corneal optical anisotropies in a rabbit model of limbal stem cell deficiency
title_fullStr Clinical features and corneal optical anisotropies in a rabbit model of limbal stem cell deficiency
title_full_unstemmed Clinical features and corneal optical anisotropies in a rabbit model of limbal stem cell deficiency
title_sort Clinical features and corneal optical anisotropies in a rabbit model of limbal stem cell deficiency
author Kobashigawa, Karina Kamachi [UNESP]
author_facet Kobashigawa, Karina Kamachi [UNESP]
Aldrovani, Marcela [UNESP]
Barros Sobrinho, Alexandre A. F. [UNESP]
Santo Silva, Paloma do Espirito [UNESP]
Marcusso, Paulo F. [UNESP]
Marinho-Neto, Fausto A. [UNESP]
Martines Padua, Ivan R. [UNESP]
Laus, Jose Luiz [UNESP]
author_role author
author2 Aldrovani, Marcela [UNESP]
Barros Sobrinho, Alexandre A. F. [UNESP]
Santo Silva, Paloma do Espirito [UNESP]
Marcusso, Paulo F. [UNESP]
Marinho-Neto, Fausto A. [UNESP]
Martines Padua, Ivan R. [UNESP]
Laus, Jose Luiz [UNESP]
author2_role author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Universidade Estadual Paulista (Unesp)
dc.contributor.author.fl_str_mv Kobashigawa, Karina Kamachi [UNESP]
Aldrovani, Marcela [UNESP]
Barros Sobrinho, Alexandre A. F. [UNESP]
Santo Silva, Paloma do Espirito [UNESP]
Marcusso, Paulo F. [UNESP]
Marinho-Neto, Fausto A. [UNESP]
Martines Padua, Ivan R. [UNESP]
Laus, Jose Luiz [UNESP]
dc.subject.por.fl_str_mv Birefringence
Corneal stroma
Anisotropy
Stem cells
Limbus corneae
Glycosaminoglycans
topic Birefringence
Corneal stroma
Anisotropy
Stem cells
Limbus corneae
Glycosaminoglycans
description Purposes: To investigate the intra-laboratory reproducibility of clinical features and to evaluate corneal optical anisotropies in a rabbit model of limbal stem cell deficiency. Methods: Limbal injury was induced in the right eye of 23 adult New Zealand White rabbits using a highly aggressive protocol that combined 360 degrees limbal peritomy, kerato-limbectomy, alkaline chemical burn, and mechanical removal of the epithelium. Clinical evaluation of the injured eyes was performed for 28 days and included corneal impression cytology. Corneas with a severe clinical outcome set typical of limbal stem cell deficiency were then collected, subjected to a histopathological examination, and examined for optical anisotropies. Corneas from healthy rabbit eyes were used as controls. Differences in optical path due to stromal collagen birefringence, as well as linear dichroism related to the expression and spatial orientation of glycosaminoglycan chains from proteoglycans, were measured from cross-sections under a quantitative polarized light microscope. Results: One eye showed signs of hypopyon and was excluded. Signs of ocular inflammation were observed in all eyes studied (n=22). Corneal impression cytology did not detect goblet cells. Twelve of the 22 corneas presented a clinical outcome set typical of limbal stem cell deficiency, which is characterized by the presence of epithelial defects, inflammatory cells, moderate-to-severe opacity, and neovascularization. Microscopic studies under polarized light revealed that relative to controls, limbal stem cell deficiency caused a 24.4% increase in corneal optical path differences. Further, corneas with limbal stem cell deficiency were less dichroic than controls. Conclusions: These results suggest that rabbit models of limbal stem cell deficiency must be rigorously screened for use in preclinical studies to ensure experimental homogeneity because protocols used to create limbal stem cell deficiency could be not associated with good intra-laboratory reproducibility of clinical features. Limbal stem cell deficiency, as induced herein, altered the optical anisotropic properties of the corneal stroma. Such alterations are indicative of changes in collagen packing and the spatial orientation of glycosaminoglycan chains from proteoglycans. Knowledge of these changes is important to potentiate strategies aimed at restoring the morphofunctional integrity of the corneal stroma affected by limbal stem cell deficiency.
publishDate 2018
dc.date.none.fl_str_mv 2018-11-26T17:55:20Z
2018-11-26T17:55:20Z
2018-09-01
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://dx.doi.org/10.5935/0004-2749.20180076
Arquivos Brasileiros De Oftalmologia. Sao Paulo: Consel Brasil Oftalmologia, v. 81, n. 5, p. 384-392, 2018.
0004-2749
http://hdl.handle.net/11449/164620
10.5935/0004-2749.20180076
WOS:000443858200006
WOS000443858200006.pdf
url http://dx.doi.org/10.5935/0004-2749.20180076
http://hdl.handle.net/11449/164620
identifier_str_mv Arquivos Brasileiros De Oftalmologia. Sao Paulo: Consel Brasil Oftalmologia, v. 81, n. 5, p. 384-392, 2018.
0004-2749
10.5935/0004-2749.20180076
WOS:000443858200006
WOS000443858200006.pdf
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Arquivos Brasileiros De Oftalmologia
0,518
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 384-392
application/pdf
dc.publisher.none.fl_str_mv Consel Brasil Oftalmologia
publisher.none.fl_str_mv Consel Brasil Oftalmologia
dc.source.none.fl_str_mv Web of Science
reponame:Repositório Institucional da UNESP
instname:Universidade Estadual Paulista (UNESP)
instacron:UNESP
instname_str Universidade Estadual Paulista (UNESP)
instacron_str UNESP
institution UNESP
reponame_str Repositório Institucional da UNESP
collection Repositório Institucional da UNESP
repository.name.fl_str_mv Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)
repository.mail.fl_str_mv
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