Freeze-drying as an alternative method of human sclera preservation

Arquivos Brasileiros de Oftalmologia

Ana Carolina De Arantes Frota²  Acácio Alves De Souza Lima Filho¹  Ana Beatriz Toledo Dias¹  Andréia Cristina Dos Santos Lourenço¹  Emilia Antecka¹  Miguel N. Burnier Jr.¹ 

[1] ,McGill University Department of Ophthalmology Henry C. Witelson Ocular Pathology LaboratoryMontreal,Canada

关键词: Freeze-drying;    Sclera;    Tissue preservation;    Dose-response relationship;    drug;    Ethanol;    Immunohistochemistry;    Collagen;    Liofilização;    Esclera;    Preservação de tecido;    Relação dose-resposta a droga;    Etanol;    Imunoistoquímica;    Colágeno;   

DOI  :  10.1590/S0004-27492008000200002

来源: SciELO

【 摘 要 】

PURPOSE: To compare the effect of preserving sclera samples in either 95% ethanol or freeze-dried. METHODS: Ninety-six samples of human sclera were studied. Half of them were freeze-dried and half preserved in 95% ethanol. Preservation periods of 18, 45, 90 or 174 days were studied. Automated immunostaining was carried out in the Ventana BenchMarkR LT platform using collagen 1 and fibronectin antibodies. Histological staining was also performed with hematoxilin-eosin and Masson trichrome. Samples were classified according to the degree of collagen fiber parallelism (0-2), intensity of Masson staining (0-2), and the expression of both antibodies (0-3). Friedman and Wilcoxon tests were applied to compare preservation methods and p-values below 0.05 were considered to ensure statistical significance. RESULTS: Relevant results were found in three situations: (i) Friedman's test showed better collagen fiber integrity in the freeze-dried group rehydrated after 174-days as compared to the 90-day group; (ii) Wilcoxon's test showed better collagen fiber integrity in the freeze-dried group after 18 and 174 days as compared to the ethanol group; (iii) the freeze-dried group disclosed higher immunohistochemical expression for COL-1 antibody in the sclera samples rehydrated after 45, 90 and 174 days as compared to the ones rehydrated after 18 days. CONCLUSION: Histological and immunohistochemical analysis showed freeze-drying to be a superior method for sclera preservation as compared to 95% ethanol. This technique provides an easy method to manipulate tissue, with longer shelf life, and storage at room temperature.

【 授权许可】

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