Correlation of Immunohistochemical Biomarkers and Differential Staining Techniques to Investigate the Role of Subepithelial Hyalinization in the Aggressiveness of Odontogenic Keratocyst

Sharon John; Eram Khan; Ayushi Jain; Priya Devi; Shalini Gupta

doi:10.1007/s12070-024-05044-5

Correlation of Immunohistochemical Biomarkers and Differential Staining Techniques to Investigate the Role of Subepithelial Hyalinization in the Aggressiveness of Odontogenic Keratocyst

Indian J Otolaryngol Head Neck Surg. 2024 Dec;76(6):5610-5617. doi: 10.1007/s12070-024-05044-5. Epub 2024 Sep 9.

Authors

Sharon John^{1

2}, Eram Khan¹, Ayushi Jain¹, Priya Devi¹, Shalini Gupta¹

Affiliations

¹ Department of Oral Pathology, King George's Medical University, Lucknow, UP 226003 India.
² Department of Oral & Maxillofacial Pathology and Oral Microbiology, King George's Medical University, Lucknow, India.

PMID: 39559161
PMCID: PMC11569315 (available on 2025-12-01)
DOI: 10.1007/s12070-024-05044-5

Abstract

Odontogenic Keratocysts (OKCs) are known for their aggressive behavior along with rapid expansion. Subepithelial hyalinization (SEH) is one of the causes of recurrence. The ability to predict this biological behavior histologically may help medical experts choose the best course of action. To investigate the aggressiveness of odontogenic keratocyst caused by SEH and its recurrence tendency in the north Indian population, this study will link differential staining methods with immunohistochemistry biomarkers that can be used in routine investigative procedures. Consequently, the evaluation and grading of SEH were established by measuring from the basement membrane to the extent of connective tissue. The levels were correlated to Ki67, Alcian blue, and O -safranine for validation. Forty OKCs were examined for the histological investigation of SEH using the immunohistochemical marker Ki67 and differential staining with O-safranine and Alcian Blue. The histological trait of separation of epithelium from the connective tissue interface due to SEH was noted. SEH-positive cases that were evaluated with Ki67, had increased proliferative activity. The differential staining techniques were validated with Ki67, cross-tabulations in SPSS, and kappa statistic value was given to analyze the results. Spearman's rank correlation was done between Ki67 vs Alcian blue and O-Safranine. A p value of less than < 0.05 was considered statistically significant. In SEH-positive cases, a higher proliferative index was observed. Additionally, histological metrics were statistically significantly higher in SEH-positive cases. Consequently, SEH is a reliable histopathological indicator in OKC for predicting recurrence. The presence of SEH indicates that OKCs are more likely to recur.

Keywords: Alcian Blue; O-safranine; Odontogenic Keratocysts; Subepithelial hyalinization.

© Association of Otolaryngologists of India 2024. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.