Diffusion of reduced arecoline and arecaidine through human vaginal and buccal mucosa.

Because alkaloids from areca nut, arecoline and arecaidine, have been implicated in the development of oral submucous fibrosis, we determined their diffusion kinetics through human buccal and vaginal mucosa. Four clinically healthy vaginal mucosa specimens (mean patient age +/- standard deviation: 47 +/- 15 years; age range: 31-60 years) and 4 buccal mucosa specimens from 2 male patients and 2 female patients (mean patient age +/- standard deviation: 31 +/- 9 years; age range: 17-53 years) were obtained during surgery. In vitro flux rates of reduced arecoline and arecaidine (r-arecoline and r-arecaidine) were determined by use of a flow-through diffusion apparatus. Analysis of variance, a Duncan multiple range test, and an unpaired t-test were used to determine steady state kinetics and flux differences over time intervals. Although statistically significant differences were observed between flux values for both alkaloids and tissues at certain time points, these were not considered to be of biological (clinical) significance. However, the flux rates across both mucosa of r-arecoline were significantly higher statistically than those of rarecaidine. The findings demonstrated the differences in the diffusion kinetics between r-arecoline and r-arecaidine across human buccal and vaginal mucosa, an observation that could be explained in terms of their ionisation characteristics. Additionally, the results obtained further support the hypothesis that human vaginal mucosa can be used as a model for buccal mucosa in studies of permeability to various chemical compounds.

Oral mucosal lesions associated with betel quid, areca nut and tobacco chewing habits: consensus from a workshop held in Kuala Lumpur, Malaysia, November 25-27, 1996.

A variety of betel/areca nut/tobacco habits have been reviewed and categorized because of their possible causal association with oral cancer and various oral precancerous lesions and conditions, and on account of their widespread occurrence in different parts of the world. At a recent workshop in Kuala Lumpur it was recommended that "quid" be defined as "a substance, or mixture of substances, placed in the mouth or chewed and remaining in contact with the mucosa, usually containing one or both of the two basic ingredients, tobacco and/or areca nut, in raw or any manufactured or processed form." Clear delineations on contents of the quid (areca nut quid, tobacco quid, and tobacco and areca nut quid) are recommended as absolute criteria with finer subdivisions to be added if necessary. The betel quid refers to any quid wrapped in betel leaf and is therefore a specific variety of quid. The workshop proposed that quid-related lesions should be categorized conceptually into two categories: first, those that are diffusely outlined and second, those localized at the site where a quid is regularly placed. Additional or expanded criteria and guidelines were proposed to define, describe or identify lesions such as chewer's mucosa, areca nut chewer's lesion, oral submucous fibrosis and other quid-related lesions. A new clinical entity, betel-quid lichenoid lesion, was also proposed to describe an oral lichen planus-like lesion associated with the betel quid habit.

The fibroblast population in oral submucous fibrosis.

The purpose of the investigation was to compare the morphology of fibroblasts cultured from healthy oral mucosa and mucosa of patients with oral submucous fibrosis (OSF) and to collate the occurrence of cell types of similar morphology. Cells cultured from biopsy specimens from the buccal mucosa of six subjects who did not chew the areca nut and six patients with OSF who chewed areca nut were grown according to standard techniques. Ninety cells per cell line were recorded daily for 8 days, classified into types F1, F2 and F3 according to their morphology, and the results statistically analyzed. We found that there was a relative increase of F3 cells in relation to F1 cells in OSF, resulting in the ratio of F3 to F1 cells being significantly larger in OSF than the ratio in the controls. As it has been reported that F3 cells in rat connective tissues produce significantly more collagen types I and III than F1 cells, we concluded that a change of fibroblast population has occurred in OSF and that this relative increase of F3 cells in humans, which could be committed to the production of large quantities of collagen, can be an explanation for the excessive collagen formation in OSF.

The effect of areca nut on salivary and selected oral microorganisms.

The aim of this study was to investigate the effect on the growth of salivary and selected oral microorganisms of areca nut, aqueous extracts of the nut, its major alkaloid arecoline and the components tannic acid and catechin of its tannin fraction. The antibacterial properties of the above were tested on Streptococcus mutans, Streptococcus salivarius, Candida albicans and Fusobacterium nucleatum and, as a control, Staphylococcus aureus. This was followed by investigating its effect on salivary organisms cultured from the saliva after chewing boiled areca nut. Extracts inhibited the growth of the selected organisms in a concentration dependent manner, baked and boiled nuts being significantly more potent than raw nut. Growth of C. albicans was the least affected by the nut extracts. Tannic acid was strongly antibacterial but not catechin or arecoline. No antibacterial effect could be demonstrated on salivary organisms after chewing the nut for 5 minutes but exposure of saliva to the cud for 1 hour caused a significant depression of bacterial growth. It is concluded that the hydrolysable tannins in the tannin fraction, which include tannic acid, are responsible for the antibacterial properties of the nut and that prolonged intraoral exposure to the nut can suppress bacteria in the mouth.

The effect of baked areca nut extract on the growth of buccal mucosa fibroblasts from healthy non-areca nut chewers.

Oral submucous fibrosis is a serious oral disease associated with the habit of chewing areca nut. The habit is relatively common among South Africans of Indian descent. The aim of this study was to observe the effect on growth of buccal mucosa fibroblasts derived from healthy individuals not practising the areca nut habit when the cells are exposed to nut extract. Fibroblast cell-lines from 6 individuals were grown in medium without extract and medium containing 50 and 100 [symbol: see text] g/ml extract of baked nut for 8 days. Cells not exposed to the nut extract behaved homogeneously. Reaction to the nut extracts, however, was dissimilar. The cells of 5 individuals showed no discernable reaction to the extracts while in one instance, the cells of a healthy adult male with no physical disabilities, showed marked growth inhibition. Thus, the finding indicates that when the effect of the nut or its constituents are tested on cells, it is necessary to use several cell-lines of the same cell type or a cell-line of which the growth parameters are standardized.

Growth of oral and skin fibroblasts from patients with oral submucous fibrosis.

The purpose of the investigation was to evaluate and compare the proliferation (growth) of mouth fibroblasts and skin fibroblasts from patients with oral submucous fibrosis (OSF). Material comprised fibroblasts from fibrous bands situated in the buccal mucosa and from the inner aspect of the forearm of 8 patients with classic features of OSF as well as fibroblasts from 6 buccal mucosa and 8 skin biopsy specimens from healthy non-areca nut chewing individuals. Cells were cultured for 8 days according to standard techniques. Their growth was monitored daily, under optimal conditions as well as exposure to concentrations of arecoline. The data were analyzed using regression analysis, analysis of variance and the Kruskal-Wallis test. We found no statistically significant differences between the proliferation patterns of oral and skin fibroblasts from patients or between those from patients and controls. The reaction of the cells exposed to concentrations of arecoline was similar; at low concentrations (0.1-10 micrograms/ml) normal growth was maintained, while 100 micrograms/ml inhibited growth. It is concluded that fibroblasts from mouths affected by OSF have proliferation patterns which fall within normal parameters, that the excessive collagen formation in established OSF is not due to increased fibroblast proliferation and that arecoline does not stimulate fibroblast proliferation.

Effect of chilli (Capsicum frutescens) extract on proliferation of oral mucosal fibroblasts.

Various concentrations of chilli extract were tested in 2 stages on the growth of a human buccal mucosa fibroblast cell line. Firstly, concentrations of 25, 75, 150 and 300 micrograms/ml were tested on the cell line for 6 days followed by a repeat which included concentrations of 400 and 500 micrograms/ml. In the latter, cell growth was monitored for 18 days. From day 3 (72 hr) suppression of cell growth was evident with concentrations 300-500 micrograms/ml. Total cell death occurred at 16 days with 300 micrograms/ml and at 6 days with 400-500 micrograms/ml. With the lower concentrations, 25-150 micrograms/ml, the daily counts were lower than the control but the difference was not statistically significant. Growth continued unabated. It is therefore concluded that cytopathic effect of chilli extract to fibroblasts is concentration dependent.

Observations on the effect of areca nut extracts on oral fibroblast proliferation.

The effects of aqueous extracts of raw, baked and boiled areca nuts were tested on cultured human buccal mucosa fibroblasts. Cells were exposed to extract concentrations of 0, 50, 100, 150, 300 and 500 micrograms/ml. The arecoline and arecaidine content was determined in the extracts with HPLC and raw nut contained 5.5% m/m, baked nut 6.6% m/m and boiled nut 7.1% m/m. Extract concentrations of 50 to 150 micrograms/ml inhibited cell growth in a concentration-dependent manner but did not lead to total cell death during a 7 day period. However, total cell death did occur with concentrations of 300 and 500 micrograms/ml. It is concluded that areca nut extract is toxic to cultured fibroblasts and inhibits their proliferation in a concentration-dependent manner.

HLA-antigens in oral submucous fibrosis.

HLA-typing was carried out on 122 areca nut chewers who attended hospitals for complaints unrelated to the habit. The subjects were South Africans of Indian extraction. The study did not include haplotypes. Palpable fibrous bands in the mouth indicated oral submucous fibrosis. The subjects were divided into 4 groups based on specific oral symptoms and signs. Groups A and B were without fibrous bands. Group A (47 subjects) included those with one or no symptoms while group B (28 subjects) suffered from 2 to 7 oral symptoms. Group C (17 subjects) had oral symptoms and represented early or mild oral submucous fibrosis and exhibited at least one discrete palpable fibrous band. Group D (30 subjects) were classic oral submucous fibrosis cases with multiple bands. The high occurrence of oral submucous fibrosis in this study group (39%) is similar to the occurrence in comparable age groups reported earlier in South Africa and is conceivably due to the higher age range of the subjects and their relatively long exposure to the areca nut. We were unable to demonstrate a specific pattern of HLA-antigen frequencies in chewers with or without the disease. Furthermore, there were no differences between the study population and the controls. It is concluded that there is not necessarily a HLA-associated susceptibility in oral submucous fibrosis.

The areca nut chewing habit and oral squamous cell carcinoma in South African Indians. A retrospective study.

A retrospective study (1983-1989) of oral squamous carcinomas and concomitant oral habits was undertaken in South African Indians from Natal. Information came from hospital records and interviews with patients, families and friends. There were 143 oral squamous carcinomas; these occurred in a ratio of 1:1,6 for men and women respectively. Squamous carcinomas of the cheek (buccal mucosa, alveolar sulcus and gingiva) occurred most frequently, especially in women (57/89--64%), while in men tongue cancer predominated (22/54--41%). Ninety-three per cent of women (83/87) and 17% of men (9/54) habitually chewed the areca nut. Thirty-nine of 57 women (68%) with cheek cancer and 21/25 (84%) with tongue cancer only chewed the nut (no tobacco, snuff or smoking). Analyses confirmed an association between nut chewing and cheek cancer. The odds ratio (OR) for oral cancer in women 25 years and older who only chewed the nut was 43,9 and the attributable risk (AR) 0,89 (89%). With tobacco the OR increases to 47,42 and the AR to 0,91 (91%). The data showed that the areca nut habit with or without tobacco use is important in the development of oral squamous carcinoma. Elimination of this habit can reduce the risk in these women substantially (89-91%) if all other factors remain the same.

Toothbrushing with the root of Euclea natalensis.

The custom of cleaning teeth and the gums with a chewed root of the tree Euclea natalensis, in the belief that it benefits oral health, is practised in South Africa by married women of an African Zanzibari community. The root of E. natalensis contains naphthoquinones which are bactericidal and, as they are also pigments, the mouths of the root users are stained bright orange. Samples of fresh root were tested against Streptococcus mutans and organisms in saliva and from deep periodontal pockets. Aerobic as well as anaerobic bacterial growth was suppressed in all instances. In addition we compared the plaque index (Silness et Löe), the periodontal index (CPITN), the DMFT and the state of the oral mucosa of root users and non-users. No significant differences were recorded in any of the variables. Although the beneficial effects claimed for this custom may have scientific validity, it is concluded that the daily practice is too limited to have a demonstrable beneficial effect.

Betel-nut chewing and submucous fibrosis in Durban.

A stratified random sample among South African Indians living in Durban revealed the occurrence of betel-nut chewing and the resultant submucous fibrosis (SF) to be relatively high. Women chewers predominated in a ratio of 13:1. The habit increased with age and 30.6% of women over 65 years practised the custom. Thirty-eight per cent of chewers revealed signs of impending and established SF; women predominated 70:1, and the majority of sufferers in this instance belonged to the age group 45-54 years (12.9%). Forty-six per cent of those with signs associated with SF had fibrous bands in the mouth and were regarded as having established SF. It was calculated that 5% of the total Indian population in South Africa could be chewers and that 2.3% may develop SF.

Betel chewing and dietary habits of chewers without and with submucous fibrosis and with concomitant oral cancer.

The betel-nut chewing habit and the intake of chillies in the diet were analysed in 178 chewers identified during a stratified random survey and in 124 hospital patients suffering from submucous fibrosis (SF). There were no signs or symptoms of SF in 115 chewers from the survey; the remaining 63 exhibited features of impending and established SF. Of the 124 hospital patients, 10 also had oral cancer. Subjects suffering from SF had practised the habit in whatever form for a significantly shorter period than chewers without SF and belonged to a significantly younger age group. A significantly larger proportion of this group preferred the boiled nut by itself and not as part of the betel package (paan or pan). No relationship could be established between SF and the use of tobacco, lime or chillies. There was no difference in frequency of the habit between chewers without SF and chewers with SF. The 10 oral cancer patients all had moderately to well-differentiated squamous carcinomas arising in the buccal sulci (8), the soft palate (1) and upper oesophagus (1). Eight had cancer on presentation and 2 developed the tumour while under observation. Nine chewed the nut only and only 1 preferred paan which included tobacco, lime and catechu, which is contrary to the proposal that tobacco or lime are the carcinogens. It is concluded that a relationship between betel-nut chewing and SF exists but the mechanism by which the disease develops is still obscure. A genetic predisposition may be important.

The oral features of betel nut chewers without submucous fibrosis.

This study examines the betel nut chewing practices, and the resulting oral clinical and histological features of 20 chewers who were without the diagnostic characteristics of submucous fibrosis. All chewed pan; the average duration of the habit was 10.5 years and the average frequency 7.1 quids per day. Eleven complained of intermittent burning and soreness of the mouth and 12 depicted blanching of the mucosa. Only 2 subjects were free from symptoms or signs. Eighteen chewers revealed an abnormal epithelium with extensive hyperplasia in 10 subjects and the rest demonstrated varying degrees of hyperplasia with abnormal maturation of the epithelium. Seven of the latter showed atypia as well. Parakeratosis was present in 7 and pigment incontinence in 9. Varying degrees of chronic inflammation featured in 17 specimens and the lamina propria was dense and hyalinized in 9. No correlation was found between the histological features, the clinical features and the various aspects of the habit. It is concluded that chewing betel nut will lead to pathological changes in the mucosa, but that neither the frequency nor the duration of the habit are accurate predictors of the extent of these changes or when they are likely to occur.

Submucous fibrosis (SF) in ex-betel nut chewers: a report of 14 cases.

The clinical and histological features are described of 14 subjects with SF who had given up betel nut chewing. The typical clinical features of the disease: blanching of the mucosa, palpable fibrous bands and complaints of burning and dryness of the mouth were still present, even in subjects who had not practised the habit for as long as 13 years. The characteristic histological features of the disease: atrophy of the oral epithelium and extensive fibrosis of the lamina propria, which in some instances penetrated into the submucosa, also persisted.

Submucous fibrosis in non-betel nut chewing subjects.

Six patients with the typical features of submucous fibrosis (SF) but without a history of betel nut chewing or an abnormal intake of chillies are described. All had palpable fibrous bands in their mouths and limited mouth opening as well as a variety of symptoms associated with SF. Biopsy specimens of the cheeks showed the classic features of SF, an atrophic epithelium and a very dense lamina propria. Also seen were hyperkeratosis, atypia and signet cell-like degeneration of the epithelium and chronic inflammation of the lamina propria. In 3 cases the fibrosis extended into the submucosa. As no reason for the disease could be demonstrated the question is asked whether people with a genetic predisposition can develop SF spontaneously.

Calcium, phosphorus, fluoride, and pH levels of human dental plaque from areas of varying fluoride levels.

Plaque calcium, phosphorus, fluoride, and pH in samples obtained from 149 life-long resident children, aged from six to seven and 12 to 13 yr, from one of three naturally fluoridated communities were determined. In general, smaller amounts of phosphorus and fluoride, and larger amounts of calcium and hydrogen ions (lower pH values) are associated with smaller amounts of fluoride in the drinking water supply. Significant differences among the mean pH, fluoride, and Ca/P concentrations both in the six-to-seven- and 12- to 13-year age group were found when comparing the results of the three areas with different levels of fluoride in their drinking water supply.

Oral lesions caused by habits.

Oral lesions caused by habits can be of teeth only, of teeth and the soft tissues of the mouth or may be only of the soft tissues. Lesions of teeth are permanent and may remain even when there has been total destruction of soft tissues. Recognition of lesions due to habits such as betel chewing, snuff dipping, pipe smoking and certain sexual practices may help towards establishing the sex, the ethnic grouping or even the place of origin of a person or their remains. Certain dental and gingival changes may indicate a person's working circumstances and even the type of treatment received.