Comparison between the techniques of inclusion in glycol methacrylate (GMA)-based plastic resin and paraffin for evaluation intestinal morphometry in horses.
Abstract: To determine the effect of the inclusion method on the histomorphometric evaluation of the gastrointestinal mucosa of horses, jejunum samples were collected using flank laparotomy. Sixteen mixed breed healthy adult horses, including four males and 12 females, aged 4-14 years with an average body weight of 248.40 ± 2.28 kg, were used. Jejunal biopsies were collected and analyzed by light microscopy using two methods: group 1 comprised biopsies fixed using 10% neutral formalin and embedded in paraffin; biopsies in group 2 were fixed in 2.5% glutaraldehyde in 0.1 M phosphate buffer pH 7.2, followed by inclusion in glycol methacrylate (GMA)-based plastic resin. Intestinal villi height, crypt depth, glandular mucosa thickness, total mucosal thickness, and villus/crypt ratio were then evaluated. For all the variables studied, with exception of the villus/crypt ratio, a significant difference (p < 0.05) was found between samples in groups 1 and 2. Processing samples for embedding in plastic resin was quicker and easier to perform compared to that for paraffin embedding. In addition, the epithelial lining of tissues in group 2 showed better resolution for conducting cytological studies under a light microscope. The difference between the studied variables can be attributed to tissue retraction caused by conventional processing for inclusion in paraffin. Therefore, the method of inclusion in GMA described in the present study appears to be a more reliable choice for morphometric evaluation of the intestinal mucosa of horses. Para determinar o efeito do método de inclusão sobre a avaliação histomorfométrica da mucosa gastrointestinal de equinos foram coletadas amostras do jejuno por laparotomia pelo flanco. Foram utilizados 16 equinos adultos hígidos, sem raça definida, de ambos os sexos, quatro machos e 12 fêmeas, com idade variando entre quatro e 14 anos, e peso corporal médio de 248,40 2,28kg. Amostras do jejuno foram coletadas e processadas para biopsia em microscopia de luz sob dois métodos: grupo 1 - fixação em formalina neutra tamponada a 10% e inclusão em parafina, grupo 2 - fixação em glutaraldeído 2,5% em tampão fosfato 0,1M pH 7,2, seguido de inclusão em resina plástica à base de glicol metacrilato. Os seguintes parâmetros foram avaliados: altura das vilosidades intestinais, profundidade de cripta, espessura da mucosa glandular, espessura total da mucosa e relação vilo/cripta. Para todas as variáveis estudadas, exceto relação vilo/cripta, foi encontrada diferença significativa (p<0,05) entre os dois métodos. O processamento para inclusão em resina plástica foi rápido e de fácil execução quando comparado à inclusão em parafina. Além disso, o epitélio de revestimento apresentou melhor resolução das células para estudo histológico ao microscópio de luz. A diferença entre as variáveis pode ser atribuída a retração do tecido provocada pelo processamento convencional por inclusão em parafina. Portanto, o método de inclusão em GMA mostrou-se, no presente estudo, uma escolha mais fidedigna para as avaliações morfométricas da mucosa intestinal de equinos.
Copyright Ferreira et al.
Publication Date: 2022-04-12 PubMed ID: 35749088PubMed Central: PMC9179190DOI: 10.29374/2527-2179.bjvm004521Google Scholar: Lookup
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- Journal Article
Summary
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The research article discusses a comparison between techniques of embedding biological sample sections in glycol methacrylate (GMA)-based plastic resin and paraffin when evaluating intestinal morphometry in horses. The results found that GMA-based plastic resin embedding proved to be a more reliable choice for morphometric evaluation of the intestinal mucosa of horses.
Study Design and Subjects
- The study utilized sixteen mixed breed adult horses, including four males and 12 females aged between 4 and 14 years. The average body weight among the subjects was approximately 248.40 ± 2.28 kg.
Methods of Data Collection
- Jejunal biopsies were collected from the horses via flank laparotomy, a surgical method. The extracted biopsy samples were then fixed and embedded in two different ways, forming two distinct study groups.
- Group 1’s samples were fixed using 10% neutral formalin and embedded in paraffin. Group 2’s samples were fixed in 2.5% glutaraldehyde in 0.1 M phosphate buffer pH 7.2, and then included in glycol methacrylate (GMA)-based plastic resin.
Data Analysis and Findings
- Each sample’s intestinal villi height, crypt depth, glandular mucosa thickness, total mucosal thickness, and villus/crypt ratio were evaluated.
- A significant difference was noted between the two groups’ samples regarding all studied variables, except for the villus/crypt ratio.
- The study found that embedding samples in plastic resin (GMA) was quicker and easier than embedding in paraffin. It further adds that group 2’s epithelial tissue lining showed better resolution for cytological studies under a microscope.
- The difference between the metrics was attributed to tissue retraction caused by the conventional processing for embedding samples in paraffin.
Conclusion
- The research concluded that the method of inclusion in GMA-based plastic resin was more reliable for morphometric evaluations of the intestinal mucosa of horses compared to paraffin embedding. This finding can influence future histological studies in equine veterinary science.
Cite This Article
APA
Ferreira C, Palhares MS, de Melo UP, Chiarini-Garcia H, Maranhão RPA, Gheller VA, Leme FOP.
(2022).
Comparison between the techniques of inclusion in glycol methacrylate (GMA)-based plastic resin and paraffin for evaluation intestinal morphometry in horses.
Braz J Vet Med, 44, e004521.
https://doi.org/10.29374/2527-2179.bjvm004521 Publication
Researcher Affiliations
- Veterinarian, DSc. Faculdade Maurício de Nassau (Uninassau). Campus Natal, RN, Brazil.
- Veterinarian, DSc. Programa de Pós-Graduação em Ciência Animal, Departamento Clínica e Cirurgia Veterinárias. Escola de Veterinária, Universidade Federal de Minas Gerais (UFMG). Campus Pampulha, MG. Brazil.
- Veterinarian, DSc. Programa de Pós-Graduação em Ciência Animal, Departamento Clínica e Cirurgia Veterinárias. Escola de Veterinária, Universidade Federal de Minas Gerais (UFMG). Campus Pampulha, MG. Brazil.
- Veterinarian, DSc. Instituo de Ciências Biológicas, UFMG. Campus Pampulha, MG. Brazil.
- Veterinarian, DSc. Instituo de Ciências Biológicas, UFMG. Campus Pampulha, MG. Brazil.
- Veterinarian, DSc. Instituo de Ciências Biológicas, UFMG. Campus Pampulha, MG. Brazil.
- Veterinarian, DSc. Instituo de Ciências Biológicas, UFMG. Campus Pampulha, MG. Brazil.
Conflict of Interest Statement
Conflict of interests: There were no conflicts of interest in carrying out this study or in the publication of its results.
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Citations
This article has been cited 1 times.- Melo UP, Palhares MS, Ferreira C, Leme FOP, Gheller VA. Effects of total parenteral nutrition associated with glutamine, enteral fluid therapy with or without glutamine, and fluid therapy on the acid-base and electrolyte balance of horses starved after exploratory laparotomy.. Braz J Vet Med 2022;44:e003222.
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