Effect of Areca Nut on Cultured Human Bucccal Fibroblast: A Cell Culture Study

Yakob, Martin (2010) Effect of Areca Nut on Cultured Human Bucccal Fibroblast: A Cell Culture Study. Masters thesis, Ragas Dental College & Hospital, Chennai.


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Betel quid (BQ) chewing is a popular habit in Taiwan, India and many Southeast Asian countries. It has been estimated that, worldwide, around 600 million people chew areca nut. Areca nut popularly known as betel nut accounts for a major portion, by weight of the betel quid, which is composed of betel leaf, areca nut, catechu and lime to which tobacco may or may not be added. Condiments, sweetening agents and spices maybe added according to individual preferences. Betel leaf is perishable and BQ preparation is complex, and this led to the emergence of commercial areca nut products about three decades ago. In India, areca nut is chewed as such or in the form of commercial products such as scented supari, manipuri tobacco, pan masala (areca quid, catechu, lime, flavors and spice), gutka or mawa (areca quid and tobacco). Oral submucous fibrosis is a pre-malignant and crippling condition of the oral mucosa characterized by submucosal fibrosis leading to atrophic changes in the epithelium resulting in mucosa appearing pale or whitish, presence of palpable fibrous bands, sensitivity to hot and spicy food and restricted mouth opening. Oral submucous fibrosis was first described by Schwartz in 1952 among five East African women of Indian origin. This was followed by first description of this condition in India in 1953 by Lal and Joshi. The overall prevalence of OSF has increased from 2.42 in 2002 to 6.42 per 1000 per year in 20047. Rapid onset of the disease in younger chewers has been reported in a hospital based case control study in Chennai, South India. Several studies concerning the etiologic factors of OSF have been done but not many focusing on the habitual use of different forms of areca nut. Areca nut contains several alkaloids of which arecoline is the most abundant. Many of the undesirable effects of areca nut have been attributed to arecoline because of its mutagenic activity in mammalian cells. Arecoline is also a cytotoxic agent to primary human buccal fibroblast cultures. Paradoxically, it has been experimentally observed that arecoline in lower concentrations stimulates collagen synthesis by fibroblasts in vitro. Strong association between areca nut use and OSF has been studied but not many have been done to assess the effects of alkaloids derived from products at a cellular level. Hence, the present study was done to isolate, culture and characterize fibroblasts from buccal mucosa of subjects without areca nut chewing habit and to evaluate the changes in the cultured fibroblast cells when exposed to extracts of various commercial areca nut products. AIMS AND OBJECTIVES: 1. To isolate and culture fibroblasts from the buccal mucosa of subjects who do not chew areca nut and have clinically normal appearing oral mucosa. 2. To evaluate the effects of commonly used areca nut products on the cultured fibroblast cells. 3. To evaluate and compare the following features: • The ratios between F1, F2, F3 subpopulations. • The ratios between mitotic and post-mitotic phenotypes. • The growth curve and its derivatives for each cell line. (Seeding efficiency and population doubling time). SUMMARY: 1. The present study was undertaken to evaluate the effects of areca nut on cultured human buccal fibroblasts from subjects without areca nut chewing habits. 2. Determination of F1:F2:F3 fibroblast subpopulation ratios, Mitotic: post mitotic fibroblast subpopulation ratios, growth curve with its derivatives were done to characterize the isolated fibroblasts. 3. We estimated the concentration levels of arecoline and arecaidine in a commercially available areca nut product using HPLC. 4. There was in an increase in post mitotic fibroblast and a decrease in mitotic fibroblast in the control group which was not statistically significant. 5. Post mitotic cells were not observed on exposure to low concentration of areca nut extract (0.175μg/ml) in five days of observation. 6. We found that there is an increase in F3 and F2 subpopulation (p<0.001) and a decrease in F1 subpopulation (p<0.001) within the control and study group. 7. We found that there is an increased trend in F3 and F2 subpopulations of cells in controls when compared to the study group.

Item Type: Thesis (Masters)
Uncontrolled Keywords: Areca Nut ; Cultured Human Bucccal Fibroblast ; Cell Culture study.
Subjects: DENTAL > Oral Pathology and Microbiology
Depositing User: Kambaraman B
Date Deposited: 12 May 2018 04:10
Last Modified: 12 May 2018 04:10
URI: http://repository-tnmgrmu.ac.in/id/eprint/7702

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