ABSTRACT
Background: Oral submucous fibrosis (OSMF) is graded according to various histological factors which include the epithelial changes and the connective tissue changes. These features could be identified in routine Hematoxylin and Eosin (H and E) staining in shades of pink. However, the use of special stains provides a contrast to various connective tissue components thereby aiding in better visualization of these connective tissue changes in advanced OSMF cases. Objective:To compare and evaluate muscle involvement and degeneration in advanced oral submucous fibrosis using three different stains namely, H&E, Van Gieson, and Masson's Trichrome. Material and Methods: 10 Formalin-fixed paraffin-embedded tissue sections of advanced OSMFwere stained using 3 different stains namely Hematoxylin and Eosin (H&E), Van-Gieson, and Masson trichrome. The results obtained were tabulated and statistically analyzed using Kruskal Wallis ANOVA. Results: The hyalinization and fibrosis involving the skeletal muscle were better visualized in Masson's Trichrome but were not statistically significant. The muscle degeneration in deeper areas was better visualized in Masson's trichrome when compared to the H&E and Van Gieson. Conclusion: Hematoxylin and eosin stains all the connective tissue components in various shades of pink, use of special stains bestows contrast between different components of connective tissue, thus improvising grading of OSMF. Masson's trichrome stain can be used as a single adjunct to hematoxylin and eosin stain as changes in the superficial and deeper connective tissue could be ascertained (AU)
Contexto: A fibrose submucosa oral (FSO) é classificada de acordo com vários fatores histológicos que incluem alterações epiteliais e do tecido conjuntivo. Essas características podem ser identificadas na coloração de rotina com Hematoxilina e Eosina (H&E) em tons de rosa. No entanto, o uso de colorações especiais fornece um contraste para vários componentes do tecido conjuntivo, auxiliando assim na melhor visualização dessas alterações do tecido conjuntivo em casos avançados de FSO. Objetivo: Comparar e avaliar o envolvimento e degeneração muscular da fibrose submucosa oral avançada usando três colorações diferentes: H&E, Van Gieson e Tricrômio de Masson. Material e Métodos: 10 seções de tecido fixadas em formalina e embebidas em parafina de FSO avançada foram coradas usando essas três colorações. Os resultados obtidos foram tabulados e analisados estatisticamente por meio do teste de Kruskal Wallis. Resultados: A hialinização e fibrose envolvendo o músculo esquelético foram melhor visualizadas no tricromo de Masson, mas não foram estatisticamente significativas. A degeneração muscular em áreas mais profundas foi melhor visualizada no tricrômico de Masson, quando comparado ao H&E e Van Gieson. Conclusão: Hematoxilina e Eosina coloram todos os componentes do tecido conjuntivo em vários tons de rosa. O uso de colorações especiais confere contraste entre os diferentes componentes do tecido conjuntivo, melhorando assim a avaliação da FSO. A coloração com tricrômio de Masson pode ser usada como um único adjunto para a coloração de Hematoxilina e Eosina, pois assim é possível verificar alterações superficiais e profundas no tecido conjuntivo. (AU)
Subject(s)
Oral Submucous Fibrosis , Fibrosis , Connective TissueABSTRACT
Tissue processing in histotechnique is an important procedure after specimen collection involving three main procedures namely, dehydration using alcohol, clearing using xylene, and infiltration by paraffin wax. Isopropyl alcohol, the widely used dehydrating agent is toxic and when exposed to heated isopropyl alcohol fumes, it leads to numerous health hazards. Anhydrous copper sulphate is less toxic than isopropyl alcohol and requires less amount of exposure to the chemicals. The aim of the study was to investigate the dehydrating potential of anhydrous CuSO4 as an isopropyl alcohol substitute. METHODSA descriptive study of forty paired soft tissue specimens were obtained from the Department of Oral Pathology and Microbiology and were subjected to routine histopathological tissue processing with isopropyl alcohol and anhydrous CuSO4 as dehydrating agents. Histomorphological criteria namely nuclear staining, cytoplasmic staining, artefacts, and background staining were evaluated and the scores were tabulated, and statistical analysis was done using SPSS version 20 by IBM. RESULTSOur study results showed that 40 % of alcohol dehydrated tissue samples had better nuclear staining than tissue samples dehydrated by anhydrous copper sulphate and 15 % of alcohol dehydrated tissue samples had better cytoplasmic staining than tissues dehydrated by Anhydrous CuSO4. Background staining of more than 20 % was seen in anhydrous CuSO4 dehydrated tissue specimens than alcohol dehydrated tissue specimens. Artefacts were seen in similar ratios in both alcohol (80 %) and anhydrous copper sulphate (75 %) dehydrated tissue specimens. CONCLUSIONSAnhydrous CuSO4 has superior dehydrating properties than isopropyl alcohol which lead to over-dehydration of the tissue specimens. Further studies are required to validate the findings.