Effects of application of various forms of areca nut preparations in buccal mucosa of Sprague-Dawley rats with relation to the development of oral submucous fibrosis

Background: Areca nut is the main etiological agent in the development of oral submucous fibrosis (OSF) and is consumed in different forms such as raw, roasted, boiled, and commercial preparations (pan masala, gutkha). This variation in availability of forms is closely linked to degree of affliction and clinical presentations of the disorder. Aims and Objectives: Identification and evaluation of tissue changes in rat buccal mucosa subjected to various areca nut forms (raw, roasted, and boiled), pan masala and pure arecoline solution for 36 weeks and its correlation with the development of OSF. Assessment of arecoline content in raw, roasted, boiled and commercial preparations of arecanut and its correlation to the tissue changes occurring after its administration along with development of OSF. Materials and Methods: A total of 37 healthy inbred rats were selected for the study. One rat was kept as normal (untreated) and rest divided into six groups (raw, boiled, roasted areca nut, pan masala, pure arecoline, and control group) and treated with respective extracts every alternate day for 36 weeks. At every 6 weeks interval, animals were sacrificed randomly and mucosal tissue biopsies collected and processed. hematoxylin and eosin, Van Gieson stains were used to assess the epithelium thickness, keratin thickness, fibrosis, and vascularity. Results: Significant histological changes seen in areca nut treated rats in comparison to control group. Histological changes included decreased keratin thickness, thinning of epithelium with partial or complete loss of rete ridges, consistent increase in deposition of compacted collagen fibers and decreased vascularity. Arecoline concentration was highest in raw areca nut, followed by pan masala, roasted areca nut, and boiled areca nut. Conclusion: Areca nut consumption in various forms (natural or processed) has significant detrimental effects on oral mucosa consistent with findings of OSF.