Sun, Dec 22, 2024
Volume 18, Issue 3 (May-Jun 2024)
mljgoums 2024, 18(3): 25-28 |
Back to browse issues page
Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
Shafieian R, Ebrahimzadeh-Bideskan A, Ranjbar E. Evaluation of different decalcifying agents on histochemical and immunohistochemical staining properties of canine osseous tissue. mljgoums 2024; 18 (3) :25-28
URL: http://mlj.goums.ac.ir/article-1-1599-en.html
URL: http://mlj.goums.ac.ir/article-1-1599-en.html
1- Department of Anatomy and Cell Biology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran
2- Department of Anatomy and Cell Biology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran; Applied Biomedical Research Center, Mashhad University of Medical Sciences, Mashhad, Iran
3- Department of Anatomy and Cell Biology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran; Student Research Committee, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran ,ranjbar.esmaeil.mums@gmail.com
2- Department of Anatomy and Cell Biology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran; Applied Biomedical Research Center, Mashhad University of Medical Sciences, Mashhad, Iran
3- Department of Anatomy and Cell Biology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran; Student Research Committee, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran ,
Abstract: (582 Views)
Background: The purpose of tissue processing is to strengthen the tissue and place it in a suitable environment for slicing without causing damage. In routine histopathology, decalcification of calcified tissues is a principal step before tissue processing. Thus, the aim of this study is to evaluate the effect of several decalcifying agents on morphological and antigenic preservation in canine mandibular tissue.
Methods: Four different decalcifying solutions, including 5% nitric acid (NA), 10% and 20% formic acid (FA), and 10% ethylene diamine tetraacetic acid (EDTA), were employed to remove mineral salts from specimens harvested from mandibular osseous tissues of four healthy Mongrel dogs (32 samples). The solutions were compared regarding the decalcification time, ease of tissue slicing, morphological organization after Alizarin red S staining, and osteocalcin antigenic preservation.
Results: Considering the short decalcification time and ease of sectioning, 5% NA) yielded better results than 10% and 20% FA and 10% EDTA (pH 7.4). In terms of the preservation of morphology and antigenicity of the tissue samples, 10% EDTA was found to be the most optimal solution, followed by 10% and 20% FA and 5% NA.
Conclusion: Our findings support EDTA as a highly preferred choice for the decalcification of canine osseous tissue when aiming for immunohistochemistry, despite its time-consuming nature. However, for general histological staining procedures, 20% FA and 5% NA are preferred.
Methods: Four different decalcifying solutions, including 5% nitric acid (NA), 10% and 20% formic acid (FA), and 10% ethylene diamine tetraacetic acid (EDTA), were employed to remove mineral salts from specimens harvested from mandibular osseous tissues of four healthy Mongrel dogs (32 samples). The solutions were compared regarding the decalcification time, ease of tissue slicing, morphological organization after Alizarin red S staining, and osteocalcin antigenic preservation.
Results: Considering the short decalcification time and ease of sectioning, 5% NA) yielded better results than 10% and 20% FA and 10% EDTA (pH 7.4). In terms of the preservation of morphology and antigenicity of the tissue samples, 10% EDTA was found to be the most optimal solution, followed by 10% and 20% FA and 5% NA.
Conclusion: Our findings support EDTA as a highly preferred choice for the decalcification of canine osseous tissue when aiming for immunohistochemistry, despite its time-consuming nature. However, for general histological staining procedures, 20% FA and 5% NA are preferred.
Research Article: Research Article |
Subject:
Pathology
Received: 2022/11/30 | Accepted: 2023/04/8 | Published: 2024/06/8 | ePublished: 2024/06/8
Received: 2022/11/30 | Accepted: 2023/04/8 | Published: 2024/06/8 | ePublished: 2024/06/8
References
1. González-Chávez SA, Pacheco-Tena C, Macías-Vázquez CE, Luévano-Flores E. Assessment of different decalcifying protocols on Osteopontin and Osteocalcin immunostaining in whole bone specimens of arthritis rat model by confocal immunofluorescence. Int J Clin Exp Pathol. 2013; 6(10): 1972-83. [View at Publisher] [PMID] [Google Scholar]
2. Bhaskar SN. Orban's oral histology and embriology. Orban's oral histology and embriology. 2012; 482. [View at Publisher]
3. Singh VM, Salunga RC, Huang VJ, Tran Y, Erlander M, Plumlee P, et al. Analysis of the effect of various decalcification agents on the quantity and quality of nucleic acid (DNA and RNA) recovered from bone biopsies. Annals of Diagnostic Pathology. 2013 ;17(4): 322-6. [View at Publisher] [DOI] [PMID] [Google Scholar]
4. Savi FM, Brierly GI, Baldwin J, Theodoropoulos C, Woodruff MA. Comparison of different decalcification methods using rat mandibles as a model. Journal of Histochemistry & Cytochemistry. 2017; 65(12): 705-22. [View at Publisher] [DOI] [PMID] [Google Scholar]
5. Chow DH, Zheng L, Tian L, Ho K-S, Qin L, Guo X. Application of ultrasound accelerates the decalcification process of bone matrix without affecting histological and immunohistochemical analysis. Journal of orthopaedic translation. 2019; 17: 112-20. [View at Publisher] [DOI] [PMID] [Google Scholar]
6. Gupta S, Jawanda MK, Manjunath S, Bharti A. Qualitative histological evaluation of hard and soft tissue components of human permanent teeth using various decalcifying agents-a comparative study. Journal of clinical and diagnostic research: JCDR. 2014; 8(9): ZC69. [View at Publisher] [DOI] [PMID] [Google Scholar]
7. Kapila SN, Natarajan S, Boaz K, Pandya JA, Yinti SR. Driving the mineral out faster: simple modifications of the decalcification technique. Journal of clinical and diagnostic research: JCDR. 2015; 9(9): ZC93. [View at Publisher] [DOI] [PMID] [Google Scholar]
8. Shafieian R, Matin MM, Rahpeyma A, Fazel A, Sedigh HS, Sadr-Nabavi A, et al. The effect of platelet-rich plasma on human mesenchymal stem cell-induced bone regeneration of canine alveolar defects with calcium phosphate-based scaffolds. Iranian journal of basic medical sciences. 2017;20(10):1131. [View at Publisher] [DOI] [PMID] [Google Scholar]
9. Williams TW. Alizarin red S and toluidine blue for differentiating adult or embryonic bone and cartilage. Stain Technology. 1941; 16(1): 23-5. [View at Publisher] [DOI] [Google Scholar]
10. Emans P, Bulstra S, Kuijer R. The effects of different decalcification protocols on TUNEL and general cartilage staining. Biotechnic & Histochemistry. 2005;80(3-4):111-5. [View at Publisher] [DOI] [PMID] [Google Scholar]
11. Mattuella LG, Bento LW, Vier-Pelisser FV, Araujo FB, Fossati ACM. Comparative analysis of two fixating and two decalcifying solutions for processing of human primary teeth with inactive dentin carious lesion. Revista Odonto Ciência. 2007; 22(56): 99-105. [View at Publisher] [Google Scholar]
12. Choi S-E, Hong SW, Yoon SO. Proposal of an appropriate decalcification method of bone marrow biopsy specimens in the era of expanding genetic molecular study. Journal of pathology and translational medicine. 2015; 49(3): 236. [View at Publisher] [DOI] [PMID] [Google Scholar]
13. Liu H, Zhu R, Liu C, Ma R, Wang L, Chen B, et al. Evaluation of Decalcification Techniques for Rat Femurs Using HE and Immunohistochemical Staining. BioMed research international. 2017;2017; 9050754. [View at Publisher] [DOI] [PMID] [Google Scholar]
14. Miquelestorena-Standley E, Jourdan M-L, Collin C, Bouvier C, Larousserie F, Aubert S, et al. Effect of decalcification protocols on immunohistochemistry and molecular analyses of bone samples. Modern Pathology. 2020; 33(8): 1505-1517. [View at Publisher] [DOI] [PMID] [Google Scholar]
15. Sangeetha R, Uma K, Chandavarkar V. Comparison of routine decalcification methods with microwave decalcification of bone and teeth. Journal of oral and maxillofacial pathology: JOMFP. 2013; 17(3): 386. [View at Publisher] [DOI] [PMID] [Google Scholar]
16. Sanjai K, Kumarswamy J, Patil A, Papaiah L, Jayaram S, Krishnan L. Evaluation and comparison of decalcification agents on the human teeth. Journal of oral and maxillofacial pathology: JOMFP. 2012; 16(2): 222. [View at Publisher] [DOI] [PMID] [Google Scholar]
17. Bogoevski K, Woloszyk A, Blackwood K, Woodruff MA, Glatt V. Tissue morphology and antigenicity in mouse and rat tibia: Comparing 12 different decalcification conditions. Journal of Histochemistry & Cytochemistry. 2019; 67(8): 545-61. [View at Publisher] [DOI] [PMID] [Google Scholar]
18. Senturk YECGE. Preparation Techniques of Luminal and Hard Tissues for Scanning Electron Microscopy.
Send email to the article author
| Rights and permissions | |
 |
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
Enamad
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.