Nucleolar organizer regions (AgNOR) and subepithelial vascularization as field cancerization markers in oral mucosa biopsies of alcoholic and smoking patients

Abstract

Objective: The aim of this study was to show that variations in nucleolar organizer regions (AgNOR) and the increase in subepithelial vascularization could reveal changes related to markers of field cancerization in alcoholic and smoking patients who have not yet expressed clinical or histological malignant lesions.

Study design: Quantitative variations in epithelial AgNOR and in the vascularization of the underlying connective tissue were assessed by image analysis in histologically normal biopsy specimens from alcohol drinkers and smoking patients (DS). AgNORs were evidenced by silver staining and vessel walls were labeled by immunohistochemical demonstration of the CD34 antigen. Samples of oral mucosa of nonalcoholic, nonsmoking patients (NDS) obtained during surgical procedures served as controls. Eight parameters related to number, volume, and shape of nuclei and AgNORs, and 4 parameters related to number and diameter of vascular sections were evaluated. Differences between DS and NDS groups were statistically evaluated by means of ANOVA test and posterior Bonferroni comparisons.

Results: The morphometric analysis revealed more irregular-shaped AgNORs in the superficial and suprabasal layers of the oral mucosa of DS patients. The suprabasal layers also exhibited a significantly larger number of AgNORs. The normal oral mucosa of DS patients exhibited a greater vascular density, with predominance of small-caliber blood vessels underlying the basement membrane.

Conclusion: The variations in AgNOR and epithelial vascularization would be practical biomarkers to evaluate changes underlying the augmented risk of cancerization in oral mucosa.