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Abstract

Predicting the malignant transformation risk of oral epithelial dysplasia (OED) is critical for effective patient management and cancer prevention. In this study, we performed a layer-based analysis of immunohistochemically stained OED biopsy sections to assess cell proliferation patterns layer-wise in oral epithelial dysplasia to determine if proliferation can be used as a biomarker to predict the progression of low-grade OED. The study consists of a longitudinal patient cohort of 26 progressors and 53 non-progressors, with a minimum of 5 years of follow-up for non-progressors or progression to severe dysplasia, carcinoma in situ (CIS), or oral squamous cell carcinoma (OSCC). Biopsies were digitally scanned, and the epithelial tissue was delineated into basal, parabasal, suprabasal, and higher layers. Proliferation, as indicated by Ki67 expression, was automatically quantified. Findings demonstrate that the non-progressing cases display significantly higher proliferation within the parabasal layer compared to progressing cases (p < 0.001). In analyses of layer combinations, the parabasal layer is present in every statistically significant result. In contrast, any proliferative activity in suprabasal layers and above fails to reach statistical significance. The findings suggest that the absence of normal proliferation activity in the progressing samples is linked to malignant transformation. While proliferation alone is insufficient as a predictive biomarker, the layer-specific analysis produces novel insights into OED biology and highlights potential avenues for further research.