Comparison of oral microbiota in orthodontic patients and healthy individuals.

Orthodontic treatments often include dental displacement using a fixed retainer such as braces, which may result in the accumulation of plaque that provides a suitable environment for microorganisms to cause oral infection. So, this study was designed to investigate the microbial diversity among orthodontic patients and healthy individuals. METHODS: Fifty individuals i.e. 30 orthodontic patients and 20 normal individuals were included in this study. Samples were collected during the midterm of orthodontic treatment (10-12 months). Saliva samples were collected and total DNA was isolated. Polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE) with universal primers targeting the V3 region of 16s rRNA was used to study the bacterial diversity among different orthodontic and control groups. After DGGE profile analysis, the predominant product bands from the gel were excised, cloned, and sequenced to confirm the taxonomic identity followed by its quantification by using real-time PCR with gene-specific primers. RESULTS: Both orthodontic treatment and control groups formed two distinct clustering profiles, but the Shannon-Weaver index (H') indicated greater microbial diversity in the orthodontic group (P = 0.08). Sequence analysis and real-time PCR revealed a greater number of Pseudomonas spp. in the orthodontic group, while there was no significant difference in Streptococcal spp. CONCLUSION: This study suggested alterations in the oral microbiota following orthodontic treatment would provide diagnostic tools to identify prevalent microbes associated with oral infections that may prove useful for developing future therapies.

Effect of Lactobacillus species on Streptococcus mutans biofilm formation.

Streptococcus mutans is the primary pathogen responsible for initiating dental caries and decay. The presence of sucrose, stimulates S. mutans to produce insoluble glucans to form oral biofilm also known as dental plaque to initiate caries lesion. The GtfB and LuxS genes of S. mutans are responsible for formation and maturation of biofilm. Lactobacillus species as probiotic can reduces the count of S. mutans. In this study effect of different Lactobacillus species against the formation of S. mutans biofilm was observed. Growing biofilm in the presence of sucrose was detected using 96 well microtiter plate crystal violet assay and biofilm formation by S. mutans in the presence of Lactobacillus was detected. Gene expression of biofilm forming genes (GtfB and LuxS) was quantified through Real-time PCR. All strains of Lactobacillus potently reduced the formation of S. mutans biofilm whereas Lactobacillus acidophilus reduced the genetic expression by 60-80%. Therefore, probiotic Lactobacillus species can be used as an alternative instead of antibiotics to decrease the chance of dental caries by reducing the count of S. mutans and their gene expression to maintain good oral health.

Evaluation of cytotoxicity of areca nut and its commercial products on normal human gingival fibroblast and oral squamous cell carcinoma cell lines.

Consumption of areca nut products is the most common cause of oral cancers, particularly in South Asian countries. This study evaluates the cytotoxic and necrotizing effects of areca nut and its formulations on normal human gingival fibroblasts (HGF-1) and oral squamous cell carcinoma (OSCC, CAL-27) cell lines. Identification of various carcinogens and adulterants using LC-HR-ESI-MS/MS analysis was performed in the extracts of areca nut and its products. Apart from alkaloids and flavonoids, a major adulterant, saccharin was found in all the samples of chalia (one of the most common chewing products of areca nut) in the ranges between 1.697-7.170 mg/g of the sample. Cytotoxic studies showed that most of the areca nut products were found cytotoxic to HGF-1 cells while being relatively non-cytotoxic against CAL-27 cells, rather they promote the growth of cancer cells. Our findings revealed that the components of areca nut and its products were injurious to HGF-1 cells and caused necrosis, which may attenuate HGF-1 protection toward oral epithelial cells. Moreover, the non-cytotoxic effect of these products on cancer cell lines suggests further predisposal of the habitual chewers for developing oral carcinomas. This study will give a better understanding of the hazardous effects of areca nut products.