Application of AgNOR Scores for the Evaluation of Cytological Atypia for Oral Mucosa Exposed to Toombak from a Dental Teaching Hospital, of Sudan

Abstract

Oral cancer is one of the highly prevalent cancers worldwide and a leading cause of mortality in certain regions like South-Central Asia. Oral cancer in Sudan constitutes a serious health problem, squamous cell carcinoma (SCC) is the most prevalent type of oral malignancy. The aim of this study was to assess cellular proliferative activity of clinically healthy oral mucosal epithelial cells of toombak dippers by means of AgNOR counts and cytological atypia using H&E and Pap stain, to find out the best staining results of the three stains, and to find out the most affected age group, frequency of dipping toombak per day, and the effect of snuffing duration in cellular proliferative activity. Smears were collected from normal-appearing mouth floor mucosa and tongue from 100 patients of toombak dippers, 50 non-tobacco users for negative controls and 10 cases were toombak users and they had oral cancer for positive control. AgNORs were counted in the first 50 well-fixed, nucleated squamous cells via microscope. The laboratory analysis showed that; decreasing the number of the normal cases associated with increasing to snuffing duration and to frequency of dipping per day; that the normal cytological appearance was 25 cases of snuffing duration of 2-5 years, 20 cases of snuffing duration of 6-10 years, and 4 cases of 11-15 years of snuffing. Increasing in the cases of inflammation associated with increasing to snuffing duration and to frequency of dipping per day; that the inflamed samples was 3 cases of snuffing duration of 2-5 years, 15 cases of 6-10 years of snuffing, 16 cases in 11-15 years, in 16-20 years were 8 cases, and 3 cases in above 20 years of snuffing. The cases of a typical cells appears associated in cells appears associated with increasing to snuffing duration after 11-15 years of snuffing and to increasing in frequency of dipping per day. The AgNor number like the cytological diagnosis of H&E and Pap stain that the normal count found in short snuffing duration and the abnormal count appears with the increasing in snuffing duration after 11-15 years of snuffing, as general alteration in oral mucosa appears after 5 years and more than 5 times/day, the difference was statistically significant (P value 0.01), and associated with increasing in frequency of dipping per day. In conclusion oral exfoliative cytology using Pap and H&E stain is useful in evaluation of epithelial a typia that is frequently encountered with pre-malignant and early malignant oral lesions. The results in AgNor count like the cytological diagnosis in H&E and Pap stain, the intensive exposure to toombak is a significant factor of increasing cellular  a typia. The study recommended that there is a need to standardize the cytological proliferative marker methods to provide rapid, inexpensive, simple and applicable screening test for subjects who have been identified as being at high risk for developing oral cancer, therefore the micro nuclei frequency and AgNOR counts recommended to being used with Pap and H&E stain.