Relationship between dental fluorosis, dental caries and salivary levels of Streptococcus mutans.

Background and Objective: Worldwide, dental caries is an endemic infection and a significant public health problem. Fluoride reduces caries by helping to prevent demineralisation, by remineralising early carious lesions, and by decreasing the activity and growth of Streptococcus mutans (S. mutans). Fluoride intake above the optimal levels leads to a condition known as dental fluorosis, which causes hypo-mineralisation of the tooth. Some studies have revealed that the severity of fluorosis is directly proportional to caries, but some showed opposite results. Hence, with these contradicting results, this study was undertaken to identify the relationship between different grades of dental fluorosis, dental caries and the most common cariogenic bacteria S. mutans in saliva. Materials and Methods: A total of 90 subjects within 14- to 16-year age range were selected and categorised depending on the severity of fluorosis into three groups: group I (mild), group II (moderate) and group III (severe) based on modified Dean's fluorosis index criteria (1942). Unstimulated whole saliva samples were collected using the oral rinse technique and processed for quantification of S. mutans using Mitis Salivarius Bacitracin Agar medium. The number of colony-forming units (CFUs) was determined using a colony counter and expressed as ≥102, ≥103, ≥104 and ≥105 CFUs per ml of the sample, in accordance with the scale recommended by the manufacturer (HiMedia Laboratories). The severity of the caries was measured using decayed, missing or filled teeth (DMFT). Results: Observations revealed that the overall DMFT was directly proportional to the level of S. mutans titres. It was observed that 67 to 73% of participants showed fewer colony counts (102 and 103) with minimal DMFT scores and 27 to 33% showed higher counts (104 and 105) with higher DMFT scores in all the groups. 105 CFUs of group III showed the highest mean DMFT scores (2.9) with an increased number of involved subjects than group II (2.3) and group I (1.5). In all, male participants had higher bacterial titres and DMFT scores than females. Conclusion: Fluoride in the drinking water served as anticariogenic agent regardless of the severity of fluorosis. Severe fluorosis with a higher incidence of caries and increased CFUs of S. mutans clearly indicates the importance of preventive measures and early treatment to reduce the severity of fluorosis and prevalence of dental caries.

Assessment of Cytogenetic Damage in Flexographic Workers: Micronucleus Assay in Exfoliated Buccal Epithelial Cells.

Background and Objective: Literature review revealed that chemicals used in the printing industry show an association between genotoxicity and occupational exposure. Flexography is one type of printing technique recently becoming popular because of its fast, cost-efficient, and high-quality label printing. The micronucleus (MN) is considered to be a reliable marker for genotoxic damage, and it has a close association with cancer incidences by determining the presence and the extent of the chromosomal damage. Because there are no studies on flexographic workers (FWs), this study was intended to analyze and evaluate the effect of occupational exposure on the MN frequency of buccal epithelial cells. Materials and Methods: The study comprised 100 FWs and 100 age-matched healthy controls with and without smoking habits. Buccal epithelial cells were collected from all subjects by using a cytobrush, followed by staining with Feulgen fast green. The MN frequency was recorded for each individual using the Tolbert et al. criteria. Data was statistically analyzed by using one-way analysis of variance and the posthoc test. Results: FWs with smoking habits showed a significant increase in MN frequency (1.86 ± 1.77) than workers without the habit (1.02 ± 1.08) and controls (with the habit 1.26 ± 1.33 and without the habit 0.62 ± 0.92). However, there was no significant increase of MN in FWs without the habit when compared with controls. Conclusion: This study observed the cytogenetic damage in FWs and concludes that these workers are at greater risk for genotoxicity, and the MN assay can serve as a useful biomarker.

Expression of human chorionic gonadotropin-ß in tissue specimens, saliva and urine of oral squamous cell carcinoma patients.

Background: Oral squamous cell carcinoma (OSCC) remains as one of the leading causes of death in many of the developing countries including India. Early detection helps in improving the prognosis and survival rates. Over the years, tumor markers continue to play an important role in diagnosing and monitoring cancer progression. The ectopic production human chorionic gonadotropin-ß (hCG-ß) is one such marker that is seen in various nontrophoblastic cancers and serves as a marker for tumor prognosis. Few immunohistochemical studies have shown the presence of hCG-ß in oral cancers too. The present study investigated the immunohistochemical expression, levels of hCG-ß in saliva and urine of various grades of OSCC patients and correlated it with their histopathological grading. Materials and Methods: Tissue sections of 50 histologically confirmed OSCC were subjected to immunohistochemical staining by using hCG-ß antibody (well differentiated - 21, moderately differentiated - 21 and poorly differentiated - 8). The levels of hCG-ß in saliva and urine were estimated in these individuals, by using Beckman Coulter Access 2 automated immunoassay system and comparisons drawn. Results: hCG-ß immunopositivity was seen in 8 (38%) of 21 well-differentiated, 11 (52%) of 21 moderately differentiated and 6 (75%) of 8 poorly differentiated OSCC specimens. The levels of hCG-ß in both saliva and urine were increased in poorly differentiated (0.40 and 1.19 mIU/ml) than moderately (0.3 and 0.76 mIU/ml) and well-differentiated (0.36 and 0.48 mIU/ml) OSCC patients. Conclusion: Immunohistochemical expression, salivary and urine levels of hCG-ß could serve as an independent prognostic indicator in OSCC patients.

Effect of amygdalin on oral cancer cell line: An in vitro study.

BACKGROUND: Recently, studies have concentrated on complementary medicine in treating a large number of diseases, including cancer. Unfortunately, many of these treatment methods do not provide a permanent solution, and even if they do, many do not have a scientific corroboration. A shift toward alternative medicine and natural methods is being preferred to reduce if not counter the toxic effects of the drugs that are used to treat such diseases. Studies have shown that amygdalin is a natural plant product to have an anticarcinogenic effect on many types of cancers. However, the effect of amygdalin on oral cancers has not been studied, and hence, the present study aimed to evaluate the anticarcinogenic effect of amygdalin extracted from apricots and almonds on human oral cancer cell lines. MATERIALS AND METHODS: Oral cancer cell lines (KB cell line) were used in the present study. Ethanolic extracts of apricots and almonds were prepared using Soxhlet extraction method. The antiproliferative and cytotoxic activity on KB cell line was evaluated using 3-(4,5-dimethylthiazol-2-YL)-2,5-diphenyltetrazolium bromide assay. RESULTS: Both the extracts exhibited cytotoxic and antiproliferative activity on KB cell lines. While almonds exhibited maximum efficacy at 50 µg/mL, apricots extract required 100 µg/mL concentrations. CONCLUSION: Ethanolic extracts of amygdalin from almonds and apricots were effective as an antiproliferative agent which caused apoptosis in oral cancer cell line.

Stromal changes in apparently normal mucosa of smokers and pan chewers: a multi-parametric approach

Aim: To compare and contrast the various changes in the connective tissue among patients with the habits of smoking, pan chewing and controls by using fluorescence spectroscopy, histopathology and transmission electron microscopy. Method: Thirty subjects were categorized into three groups: pan chewers, smokers and controls without any oral lesions. Fluorescence spectroscopy was carried out using FLUOROMAX-2. Excitation spectroscopy was performed at 280 and 320nm respectively and Emission spectroscopy was performed at 340 and 390nm excitation. Subsequently, histopathological evaluation and transmission electron microscopy was done for biopsies taken from test groups and controls. Results: The mean, standard deviation and test of significance of mean values between different groups for intensity-380nm, intensity–420nm and intensity – 460nm at 320 nm excitations, showed that the Mean values in group I and group II were significantly higher than the mean value in group III with a p-value of less than 0.001. Considering the histopathological parameters, the pattern of the sub-epithelial connective tissue, the presence of chronic inflammatory cells and lysis of connective tissue was significant. The ultrastructural analysis revealed the presence of epithelioid, spindle or elongated and stellate shaped fibroblasts in the connective tissue. Conclusion: In this pilot study, we could find variations in emission characteristics of various amino acids which correlated with histopathology and electron microscopy. Our study suggests that there are connective tissue changes in oral mucosa among smokers and pan chewers, though it is apparently normal in clinical presentation. These initial connective tissue changes could determine the progression of altered mucosa to a pre-cancer or cancer, which is further related to other complex interactions. Elaborate studies are required to evaluate the significance of our hypothesis.