CC BY 4.0 · Eur J Dent 2023; 17(03): 818-822
DOI: 10.1055/s-0042-1755615
Original Article

Qualitative Histological Evaluation of Various Decalcifying Agents on Human Dental Tissue

Dusit Bumalee
1   Department of Oral and Maxillofacial Pathology, Faculty of Dentistry, Mahidol University, Bangkok, Thailand
,
1   Department of Oral and Maxillofacial Pathology, Faculty of Dentistry, Mahidol University, Bangkok, Thailand
,
Khumpee Songkampol
2   Department of Oral and Maxillofacial Radiology, Faculty of Dentistry, Mahidol University, Bangkok, Thailand
,
Natchalee Srimaneekarn
4   Department of Anatomy, Faculty of Dentistry, Mahidol University, Bangkok, Thailand
,
Nakarin Kitkumthorn
3   Department of Oral Biology, Faculty of Dentistry, Mahidol University, Bangkok, Thailand
,
4   Department of Anatomy, Faculty of Dentistry, Mahidol University, Bangkok, Thailand
› Author Affiliations

Abstract

Objective Dental hard tissue is among the hardest tissue of humans because it contains high amounts of inorganic substances. This leads to difficulty in preparing histological sections for histopathological examination. Acid and chelating agents are generally used to decalcify teeth. We aimed to compare the histological quality of teeth decalcified with various calcifying agents including 5% nitric acid, 50% formic acid with 20% sodium citrate (Anna Morse solution), 10% formic acid, commercial solution, and 14.4% neutral EDTA.

Materials and Methods Freshly extracted premolar teeth were fixed and submitted for decalcification using different agents. Histological examination was qualitatively evaluated for tissue integrity and staining quality.

Results Dentin integrity of teeth decalcified with all decalcifying agents did not show any statistical differences except that with the formic acid, whereas cementum integrity decalcified with neutral EDTA showed a superior score compared with other agents. Tissue integrity and staining quality of dental pulp cells were the best decalcified with neutral EDTA or Anna Morse solution.

Conclusion Our findings demonstrated that EDTA and Anna Morse solution gave a similar efficiency in the preservation of tissue integrity while Anna Morse solution may be recommended as a decalcification agent in routine use due to the more satisfying decalcification time than EDTA.



Publication History

Article published online:
11 October 2022

© 2022. The Author(s). This is an open access article published by Thieme under the terms of the Creative Commons Attribution License, permitting unrestricted use, distribution, and reproduction so long as the original work is properly cited. (https://creativecommons.org/licenses/by/4.0/)

Thieme Medical and Scientific Publishers Pvt. Ltd.
A-12, 2nd Floor, Sector 2, Noida-201301 UP, India

 
  • References

  • 1 Sanjai K, Kumarswamy J, Patil A, Papaiah L, Jayaram S, Krishnan L. Evaluation and comparison of decalcification agents on the human teeth. J Oral Maxillofac Pathol 2012; 16 (02) 222-227
  • 2 Perin CP, Papalexiou V, Rodrigues Johann ACB, Ribeiro Mattos NH, Westphalen FH, Ditzel Westphalen VP. Evaluation of intracanal acetazolamide in late reimplanted rat teeth. Eur J Dent 2017; 11 (04) 417-421
  • 3 Zorlu FY, Darici H, Turkkahraman H. Histomorphometric and histopathologic evaluation of the effects of systemic fluoride intake on orthodontic tooth movement. Eur J Dent 2019; 13 (03) 361-369
  • 4 Pribadi N, Budiarti D, Kurniawan HJ, Widjiastuti I. The NF-kB and collagen type 1 expression in dental pulp after treated calcium hydroxide combined with propolis. Eur J Dent 2021; 15 (01) 122-126
  • 5 Lynch MJRSS. Lynch's Medical laboratory technology. Philadelphia: W.B. Saunders; 1983
  • 6 Cook SF, Ezra-Cohn H. A comparison of methods for decalcifying bone. J Histochem Cytochem 1962; 10: 560-563
  • 7 Culling CFA. Handbook of histopathological and histochemical techniques: (including museum techniques). London: Butterworth; 1974
  • 8 Mattuella LG, Frasca LC, Bernardi L, Moi GP, Fossati AC, de Araújo FB. Tooth supported prosthetic rehabilitation in a 5-year-old child with early childhood caries. J Clin Pediatr Dent 2007; 31 (03) 171-174
  • 9 Gupta S, Jawanda MK, Sm M, Bharti A. Qualitative histological evaluation of hard and soft tissue components of human permanent teeth using various decalcifying agents - a comparative study. J Clin Diagn Res 2014; 8 (09) ZC69-ZC72
  • 10 Zappa J, Cieslik-Bielecka A, Adwent MCT, Sabat D. Comparison of different decalcification methods to hard teeth tissues morphological analysis. Dent Med Probl 2005; 42 (01) 21-26
  • 11 Bhat N, Thakur K, Bhardwaj N, Singh R. Comparision of 10% nitric acid, EDTA and 10% formic acid for tooth decalcification. Adv Med Dent Health Sci 2019; 2: 26-28
  • 12 Ehrlich H, Koutsoukos PG, Demadis KD, Pokrovsky OS. Principles of demineralization: modern strategies for the isolation of organic frameworks. Part II. Decalcification. Micron 2009; 40 (02) 169-193
  • 13 Prasad P, Donoghue M. A comparative study of various decalcification techniques. Indian J Dent Res 2013; 24 (03) 302-308
  • 14 Morse A. Formic acid-sodium citrate decalcification and butyl alcohol dehydration of teeth and bones for sectioning in paraffin. J Dent Res 1945; 24 (3–4): 143-153
  • 15 Fernandes MI, Gaio EJ, Rosing CK, Oppermann RV, Rado PV. Microscopic qualitative evaluation of fixation time and decalcification media in rat maxillary periodontium. Braz Oral Res 2007; 21 (02) 134-139
  • 16 Selvig KA. Ultrastructural changes in human dentine exposed to a weak acid. Arch Oral Biol 1968; 13 (07) 719-734
  • 17 Bussolati G, Leonardo E. Technical pitfalls potentially affecting diagnoses in immunohistochemistry. J Clin Pathol 2008; 61 (11) 1184-1192
  • 18 Evans N, Krajian A. New method of decalcification. Arch Pathol (Chic) 1930; 10 (03) 447-451
  • 19 Glickman I, Wood H. Bone histology in periodontal disease. J Dent Res 1942; 21 (01) 35-54