Comparison of histochemical methods for the histological analysis of human gingival samples
Keywords:
histological staining, gum, hematoxylin, collagen, fibroblasts.Abstract
Introduction: Histological staining techniques are useful in the ultrastructural analysis of tissue samples, including gingival tissue.
Objective: Compare the usefulness of three histochemical methods (hematoxylin-eosin, Masson-Goldner and sirius red) for identification of cellular elements and other constituents of gingival tissue.
Methods: An in vitro experimental study was conducted which included the analysis of gingival tissue from healthy donors without gingival inflammation signs and indication of periodontal surgery. The gum samples were obtained by gingivectomy, processed with paraffin, cut with a microtome and placed on Polysine adhesion slides. The samples were grouped, stained with hematoxylin-eosin, Masson Goldner and sirius red, and visualized under a Leica DM 750® microscope. Reading of the findings was performed by oral pathologists.
Results: Hematoxylin-eosin staining found cellular and extracellular elements of the epithelial and connective tissue: violet-blue nuclei, pink cytoplasms, light rose collagen fibers, and arterioles and venules with differentiated tunica adventitia, media and intima. Masson-Goldner staining differentiated purple nuclei and fuchsia cytoplasm. It displayed specificity identifying green collagen fibers with dense, homogeneous and parallel distribution in the gingival connective tissue. Sirius red staining allowed identification of bright rose collagen fibers, whereas epithelial tissue and blood vessels were yellow.
Conclusion: Each of the histological staining methods evaluated in the study shows a certain affinity with and sensitivity to cellular structures and components of the specific extracellular matrix. All three are useful for the study of gingival tissue and could contribute to the analysis of gingival biopsies.
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References
Koller A, Sapra A. Anatomy, Head and Neck, Oral Gingiva. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2020 [citado 2020 Ago 20]. Disponible en: https://www.ncbi.nlm.nih.gov/books/NBK560662/
Simancas-Escorcia Víctor, Díaz-Caballero Antonio. Fisiología y usos terapéuticos de los fibroblastos gingivales. Odous Científica [Internet]. 2019 [citado 2020 Ago 20]; 20(1): 41-57. Disponible en : http://servicio.bc.uc.edu.ve/odontologia/revista/vol20n1/art05.pdf
Moutsopoulos NM, Konkel JE. Tissue specific immunity at the oral mucosal barrier. Trends Immunol [Internet]. 2018 [citado 2020 Ago 20];39(4):276–87. Disponible en: https://pubmed.ncbi.nlm.nih.gov/28923364/
Alturkistani HA, Tashkandi FM, Mohammedsaleh ZM. Histological Stains: A Literature Review and Case Study. Glob J Health Sci [Internet]. 2016 [citado 2020 Ago 20];8(3):72–9. Disponible en: http://www.ccsenet.org/journal/index.php/gjhs/article/view/48521
Wick MR. The hematoxylin and eosin stain in anatomic pathology—An often-neglected focus of quality assurance in the laboratory. Seminars in Diagnostic Pathology [Internet]. 2019 [citado 2020 Ago 20];36(5):303–11. Disponible en: https://www.sciencedirect.com/science/article/abs/pii/S0740257019300619
Salazar L, Moreno F. Comparación de tres tipos de tinciones histoquímicas en secciones histológicas de paladar y lengua de rata Wistar. SSS [Internet]. 2017 [citado 2020 Ago 20];2(2). Disponible en: https://revistas.javerianacali.edu.co/index.php/salutemscientiaspiritus/article/view/155
Lattouf R, Younes R, Lutomski D, Naaman N, Godeau G, Senni K, et al. Picrosirius red staining: a useful tool to appraise collagen networks in normal and pathological tissues. J Histochem Cytochem [Internet]. 2014 [citado 2020 Ago 20];62(10):751–8. Disponible en: https://journals.sagepub.com/doi/pdf/10.1369/0022155414545787
Rittié L. Method for Picrosirius Red-Polarization Detection of Collagen Fibers in Tissue Sections. Methods Mol Biol [Internet]. 2017 [citado 2020 Ago 20]; 1627:395–407. Disponible en: https://link.springer.com/protocol/10.1007%2F978-1-4939-7113-8_26
Feldman AT, Wolfe D. Tissue Processing and Hematoxylin and Eosin Staining. In: Day CE, editor. Histopathology: Methods and Protocols [Internet]. New York, NY: Springer; 2014 [citado 2020 Ago 20]. p. 31–43. Disponible en: https://doi.org/10.1007/978-1-4939-1050-2_3
Simancas-Escorcia VH, Carmona Lorduy M, Díaz-Caballero A. Immunolocalization of s100a4 and α-sma in gingival tissues of patients with gingival hypertrophy due to orthodontic treatment: preliminary study. Archmed [Internet]. 2021 [citado 2020 Ago 20];21(1). Disponible en: http://revistasum.umanizales.edu.co/ojs/index.php/archivosmedicina/article/view/3693
Chan JKC. The wonderful colors of the hematoxylin-eosin stain in diagnostic surgical pathology. Int J Surg Pathol [Internet]. 2014 [citado 2020 Ago 20];22(1):12–32. Disponible en: https://pubmed.ncbi.nlm.nih.gov/24406626/
Kalele K, Kulkarni N, Kathariya R. Oral Squamous Cell Carcinoma: Hematoxylin and Eosin Staining. J Clin Diagn Res [Internet]. 2015 [citado 2020 Ago 20];9(9): ZJ01. Disponible en: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4606365/pdf/jcdr-9-ZJ01.pdf
Almeida T, Valverde T, Martins-Júnior P, Ribeiro H, Kitten G, Carvalhaes L. Morphological and quantitative study of collagen fibers in healthy and diseased human gingival tissues. Rom J Morphol Embryol [Internet]. 2015 [citado 2020 Ago 20];56(1):33–40. Disponible en: https://rjme.ro/RJME/resources/files/560115033040.pdf
Kim SS, Nikoloudaki G, Darling M, Rieder MJ, Hamilton DW. Phenytoin activates Smad3 phosphorylation and periostin expression in drug-induced gingival enlargement. Histol Histopathol [Internet]. 2018 [citado 2020 Ago 20];33(12):1287–98. Disponible en: https://core.ac.uk/download/pdf/215391509.pdf
Kanneganti S, Kattappagari KK, Tanuja K, Chandra KLP, Poosarla C, Baddam VRR. Oral submucous fibrosis: Clinical and histopathological correlation of collagen fibers using Masson’s trichrome and Van Gieson stains. Journal of Dr NTR University of Health Sciences [Internet]. 2018 [citado 2020 Ago 20];7(3):181. Disponible en: http://www.jdrntruhs.org/temp/JNTRUnivHealthSci73181-263634_071923.pdf
Arora KS, Nayyar A, Kaur P, Arora KS, Goel A, Singh S. Evaluation of Collagen in Leukoplakia, Oral Submucous Fibrosis and Oral Squamous Cell Carcinomas Using Polarizing Microscopy and Immunohistochemistry. Asian Pac J Cancer Prev [Internet]. 2018 [citado 2020 Ago 20];19(4):1075–80. Disponible en: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6031770/pdf/APJCP-19-1075.pdf
Roman-Malo L, Bullon B, de Miguel M, Bullon P. Fibroblasts Collagen Production and Histological Alterations in Hereditary Gingival Fibromatosis. Diseases [Internet]. 2019 [citado 2020 Ago 20];7(2). Disponible en: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6631910/pdf/diseases-07-00039.pdf
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