The search for formalin substitute has been enduring from the time it was proven as a carcinogen. The natural sweetener, jaggery, as a tissue fixative has already been established and its compatibility with special stains demonstrated in our previous study. Hence, in the present study, the efficacy of jaggery, along with honey, was further evaluated using immunohistochemical procedures.
Materials and methods
A cross-sectional study with 30 samples of commercially available fresh goat oral mucosa, fixed with 10% buffered formalin (positive control), 20% honey, and 30% jaggery solution was conducted. After 24 hours of fixation, tissue sections were subjected to immunohistochemical stains pan-cytokeratin and desmin. Following which blinded evaluation was done by two examiners. Statistical analysis was accomplished using Kruskal—Wallis analysis of variance, Mann—Whitney U test, chi-square test, and Kappa statistics.
Results
The pan-cytokeratin staining intensity of jaggeryfixed tissues (2.67 ± 0.488) was observed to be more intense than honey (1.60 ± 0.507, p < 0.001) and formalin (2.13 ± 0.352, p = 0.003)-fixed tissues. Jaggery (2.33 ± 0.617) and formalin (2.40 ± 0.632) showed comparable results with respect to desmin.
Conclusion
Based on the findings, the fixative properties of jaggery surpassed honey and was found to be on par with formalin.
How to cite this article
Majumdar B, Rao RS, Patil S. Tissue Preservation with Natural Fixatives: An Immunohistochemical Evaluation. World J Dent 2016;7(2):87-91.
; Gamble, M. Theory and practice of histological techniques. 6th ed. Philadelphia, PA: Churchill Livingstone, Elsevier; 2008.
Chemical agents and related occupations: a review of human carcinogens. formaldehyde. Vol. 100F. Switzerland: WHO Press; 2012. p. 401-435.
Evaluation of non-formalin tissue fixation for molecular profiling studies. Am J Pathol 2002 Feb;160(2):449-457.
Basics of immunohistochemistry. J Invest Dermatol 2015 Mar;135(3):e30.
Revelation in the field of tissue preservation.a preliminary study on natural formalin substitutes. J Int Oral Health 2013 Feb;5(1):31-38.
Natural sweeteners as fixatives in histopathology: a longitudinal study. J Nat Sci Biol Med 2015 Jan-Jun;6(1):67-70.
Vimentin is preferentially expressed in human breast carcinomas with low estrogen receptor and high Ki-67 growth fraction. Am J Pathol 1990 Jan;136(1):219-227.
; Hornickel, I. Tissue fixation. the most underestimated methodical feature of immunohistochemistry. In: MndezVilas, A.; Daz, J., editors. Microscopy: science, technology, applications and education. Badajoz: Formatex Research Centre;. 2010. p. 953-959.
Chemical and physical basics of routine formaldehyde fixation. J Oral Maxillofac Pathol 2012 Sep;16(3):400-405.
Comparison of honey with ethanol as an oral cytological fixative: a pilot study. J Cytol 2015 Apr-Jun;32(2):113-117.
One of the properties of honey in wound healing is prevention of autolysis. Der Pharmacia Lettre 2010;2(3):321-325.
Review on recent advances in value addition of jaggery based products. J Food Process Technol 2015 Jan-Mar;6(4):1-2.
Jaggery - a traditional Indian sweetener. Indian J Tradit Know 2007 Jan;6(1):95-102.
; Doner, LW. Honey composition and properties: beekeeping in the United States. Agriculture Handbook No. 335; 1980. p. 82-91.
Influence of the time/temperature binomial on the hydroxymethylfurfural content of floral honeys subjected to heat treatment. Ciencia e Agrotecnologia 2012 Apr;36(2):204-209.
; Monaro, E.; Siguemoto, E.; Sefora, M. Maillard reaction products in processed food: pros and cons. In: Valdez B, editor. Food industrial processes - methods and equipment. Croatia: InTech Publishing; 2012.
Use of pharmaceutical compounds in the treatment of symptoms of disorders related to neurological diseases and etiologically related symptomology [Internet]; 1992 [Filing date 1992 Feb 21; published 1992 Sep 3; cited 2015 Oct 7]. Available from: http://www.google.co.in/patents/WO1992014456A1?cl=en
The effectiveness of honey as a substitute for formalin in the histological fixation of tissue. J Histotechnol 2006 Aug;29(3):173-176.
Honey as an alternative to formalin in the demonstration of connective tissue components. J Histotechnol 2008 Apr;31(2):67-72.
Honey as a substitute for formalin?. Biotech Histochem 2012 Feb;87(2):148-153.
Immunocytochemical evaluation of ductal carcinoma in breast after preservation in honey. J Histotechnol 2009 Mar;32(2):54-59.
Fixative properties of honey in comparison with formalin. J Histotechnol 2014 Mar;37(1):21-25.
