Assessment of dental age estimation using dentinal translucency in ground sections of single rooted teeth: a digital image analysis.

Human dentition is unique to individuals and helps in identification of individuals in forensic odontology. This study proposes to study the manually ground sections of single rooted teeth using digital methods for dental age estimation. To assess the dentinal translucency from the scanned digital images of manually ground section of teeth using commercially available image edition software. Corroborating the root dentinal translucency length and region of interest (ROI) of translucency zone in pixels (as a marker of dental age) with the chronological age of the subject, as stratified by different age groups. Twenty single-rooted extracted teeth from 20 patients each from 6 groups divided as per age. Manual sectioning of the teeth followed by scanning the sections was done. Root area in pixels and ROI of translucency zone were measured. From the observed values, translucency length percentage (TLP) and percentage of ROI in pixels (TPP) was calculated and tabulated. Pearson's correlation coefficients were obtained for age with TLP and TPP. Positive correlation existed between age and TLP and also between age and TPP. With the obtained data, multilinear regression equations for specific age groups based on 10-year intervals were derived. By a step-down analysis method, age was estimated with an average error of around ±7.9 years. This study gives a novel method for age-estimation that can be applied in real-time forensic sciences.

Diagnostic Utility of Gram Stain for Oral Smears - A Review.

For rapid and successful treatment of infectious diseases, detection of the presence of microorganisms is essential. Traditional culture-based approaches are limiting and time consuming for microbial identification. The most popular staining technique for identifying Gram-positive and Gram-negative microorganisms in various tissues is called Gram staining. This method is utilized in both clinical practice and research. Gram staining of the oral smears is the preliminary step in the identification of any pathological shift in normal oral microbiota. This review discusses the principle of gram stain emphasizing its significance in diagnostic utility for oral smears.

Radiologic-Pathologic correlation of Pindborg tumor of maxilla with extension to maxillary sinus, nasal cavity, and infraorbital margin: A case report.

Calcifying epithelial odontogenic tumor is a benign epithelial odontogenic tumor thought to originate from the stratum intermedium. Clear cell type, Langerhans cell/non-calcified type, and cystic/microcystic are the three recently recognized histological subtypes of CEOT in the 5th edition of the World Health Organization Classification of Head and Neck Tumors. Almost 350-400 cases of CEOT have been reported in literature, accounting for less than 1% of the reported cases of odontogenic tumors. We are reporting a case of Pindborg tumor of the maxilla with extension to the maxillary sinus, nasal cavity, and infraorbital margin, with an emphasis on radiographic and histopathologic presentation.

Ganglion cyst of temporomandibular joint - A systematic review.

Ganglion cyst of the temporomandibular joint (TMJ) is an uncommon pathology with uncertain etiology. There is no consensus on their management. The current systematic review aimed to discuss the clinical and histopathological features of ganglion cysts of TMJ, to aid in appropriate treatment. A literature search was done and a total of 20 cases were retrieved from published databases such as PubMed, SCOPUS, and Google Scholar. The cyst presented with swelling in all the cases followed by pain (50 %) and trismus (35 %) as other common symptoms. Though CT and MRI proved helpful in determining the location of the cyst, a histopathological examination was essential in concluding its final diagnosis. It is a pseudocyst lined by dense fibro-connective tissue with myxoid tissue degeneration. Histologically, it is essential to distinguish them from the clinically and radiographically similar true cyst of TMJ, synovial cyst. The lining of ganglion cyst is devoid of epithelium and synovial cells. Surgical excision was found to be the treatment of choice with minimal recurrence (10 %) being reported.

Inhibitory effect of garlic and omam extracts on Mucor circinelloides, a fungus causing mucormycosis: An in vitro study.

Context: Mucor circinelloides is reported to be the second among the most common causative agents of emerging mucormycosis. It is intrinsically resistant to most known antifungals. Further the use of antifungals cause side effects. Traditional knowledge system for treating various ailments is stronger in India and it also backs deriving various bioactive compounds from herbal sources, in the modern system of medicine. Therefore, two most commonly used culinary herbal materials viz., ginger and omam were studied in vitro against M. circinelloides, as an alternate to antifungal drugs. Aims: To explore the traditional herbal resources as alternate to Amphotericin B to use against M. circinelloides, a fungus causing mucormycosis. Methods and Material: Aqueous extracts of garlic and omam were prepared and tested against M. circinelloides, at different concentrations. A positive control with Amphotericin B and negative control without any supplements were also maintained. The inhibitory effect was assessed by adopting optical density (OD) measurement method in SD broth and SD Agar Well Plate using spore suspension as inoculum. Statistical Analysis Used: Paired student T test was employed using SPSS Version 1.6. Results and Conclusions: Both garlic and omam extracts were found to inhibit the M. circinelloides and their MICs were 600 and 700 µL/mL. It is comparable with the MIC of Amphotericin B, 200 µg/mL. Thus, the regular use of garlic and omam can reduce the risk of possible mucormycosis and these herbs can be explored for drug formulations against M. circinelloides.

Anti-cariogenic property of Carica papaya, Trachyspermum ammi, Caesalpinia crista linn extracts and their effect on human oral keratinocytes.

Objectives: Dental caries is a dynamic and composite process. The multifactorial etio-pathogenesis thus influences the initiation and the progression of the disease. The prime pathogenic bacterium includes Streptococcus mutans, Lactobacillus sp and Actinomyces viscosus. The purpose of this in vitro study was to analyze the antimicrobial property of the test herbal extracts and also their effects on the human oral keratinocytes. Materials and Methods: The bacterial strains S. mutans (American Type Culture Collection [ATCC]-25175); Lactobacillus species (ATCC 4356) and A. viscosus (ATCC 15987) were cultured in the specific culture media-Mitis Salivarius Bacitracin, Man Rogosa Sharpe and Enrichment media, respectively. The test extracts were exposed to the cultured plates and the mean zone of inhibition was measured. The test herbal extracts were also tested for deleterious effects on oral keratinocytes via the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Independent Student's t-test and analysis of variances were performed.-25175); Lactobacillus species (ATCC 4356) and A. viscosus (ATCC 15987) were cultured in the specific culture media-Mitis Salivarius Bacitracin, Man Rogosa Sharpe and Enrichment media, respectively. The test extracts were exposed to the cultured plates and the mean zone of inhibition was measured. The test herbal extracts were also tested for deleterious effects on oral keratinocytes via the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Independent Student's t-test and analysis of variances were performed. Results: The extracts of Carica papaya, Trachyspermum ammi and Caesalpinia crista linn inhibited the growth of bacteria and the antimicrobial effect was found to be statistically significant at the neat/standard concentration (100 µg/ml). The three extracts showed a cell viability range 96%-99% indicating that the test extracts did not produce or display any deleterious effects on the oral keratinocytes. Conclusions: The three test herbal extracts possess effective anti-cariogenic properties with near par with the efficacy of chlorhexidine and T. ammi proved to be the most potent. The extracts at different concentrations also proved to be safe, noncytotoxic producing a range of 96%-99% of cell viability of the oral keratinocytes.

Evaluation of the Anti-Carcinogenic Effect of Centella Asiatica on Oral Cancer Cell Line: In vitro Study.

OBJECTIVE: To evaluate the anti-carcinogenic effect of Centella Asiatica on to evaluate the  Anti-Carcinogenic Effect of Centella Asiatica on Oral Cancer Cell Line oral cancer cell line. MATERIALS AND METHODS:  Oral Cancer cell line and normal oral keratinocyte cell line were procured.Centella asiatica extract was prepared. The cells were then subjected to the test herbal specimens -Centella asiatica extract in succeeding concentrations of 25 µg/ml, 50 µg/ml, 100 µg/ml at time intervals of 24,48 and 72 hours. Cisplatin (2 µg/ml, 4 µg/ml, 6 µg/ml, 8 µg/ml) was used as a positive control. This experiment was done in triplets. RESULTS: The study revealed that the p values were less than 0.05 at concentration 12.5µg/ml, 25µg/ml, 50 µg/ml,100 µg/ml and time period of 24hrs,48hrs,72hrs, thus implying that at these concentrations and time period, the obtained data were statistically significant, thus indicating that there is a statistically significantly decreases in the viable cells as the concentration of the drug as a time period increases The results reveals that  centella asiatica possess potential effect of anti-carcinogenic, effect when compared to positive control (Cisplatin). CONCLUSION: The current study reveals that Centella asiatica has an potential anti-carcinogenic effect on oral cancer cell line. So this can be used to treat oral cancer with minimal crippling as compared with allopathic drugs.

Salivary stimulatory effect of novel low level transcutaneous electro neurostimulator in geriatric patients with xerostomia.

BACKGROUND: Xerostomia (dryness of the mouth) is one of the most common long-term consequences of ageing, and it causes a tremendous impact on the function and morphology of the salivary ductal system. As a consequence, it leads to a decrease in the amount of salivary output and also affects the overall quality of life. The purpose of this study was to determine whether electrostimulation using a custom designed transcutaneous electrical nerve stimulation (TENS) device will help to improve the quality of secreted saliva following electrostimulation. METHODS: One hundred thirty-five participants underwent the intervention for three months, twice daily (80 Hz). Pre-intervention and post-intervention unstimulated saliva were collected. Parameters such as salivary pH, cortisol level, salivary antioxidants, total protein, the viscosity of saliva, and microbial carriage were analysed. RESULTS: Salivary pH, cortisol, microbial cultures, viscosity, and antioxidants showed a significant difference at the end of the 3rd month (p < 0.05). Irrespective of the patient's age, gender, and common underlying systemic illnesses (diabetes and hypertension), a significant change in the quality of the salivary analytes was observed. CONCLUSION: The study emphasises the use of a custom designed TENS device in improving the quality of secreted saliva among old patients with oral dryness.

Association of MICA gene Exon-5 polymorphism in oral submucous fibrosis.

OBJECTIVE: The present study was conducted to explore the allele frequencies of MICA gene Exon-5 transmembrane and to measure the circulatory MICA levels in various histologic grades of patients with oral submucous fibrosis (OSF) compared to healthy individuals. STUDY DESIGN: We enrolled a total of 595 patients for this cross-sectional study and divided them into 2 groups: healthy controls (n = 320) and patients with OSF (n = 275). Further, patients with OSF were subdivided based on their histologic gradings. The genomic DNA was extracted followed by a polymerase chain reaction and genotyping using the ABI Prism DNA Sequencer (ThermoFisher Scientific, Inc., Waltham, MA, USA). RESULTS: Our study showed that the A5 allele of the MICA gene in the Exon-5 region conferred significant risk for patients with OSF. With reference to the histologic gradings of OSF, we found that the MICA gene conferred statistically significant risk among patients with grade III OSF. On the other hand, the A8 allele of MICA gene in the Exon-5 region conferred significant protection among the overall OSF cohort and in the grade III of histologic grade. Finally, the circulatory human MICA levels were found to have a stepwise increase from grade I toward grade III in patients with OSF. CONCLUSION: Our results suggested that the A5 allele in MICA might confer risk for the progression of OSF among the South Indian ethnic population.

Determination of Barr bodies in Transgender Patients in India - A comparative study.

Sex determination in forensic medicine is considered one of the first and foremost steps in personal identification. The need for identifying the exact sex of the individual arises when deciding whether a person can exercise certain civil rights reserved for one particular sex, for competing in sex-specific athletic and sports events, legitimacy, divorce, paternity disputes and also to some criminal offenses. Nuclear sexing by Barr body examination can be done using buccal smears to establish the sex of the individual when routine methods fail to disclose the exact gender of the individual. Aim: To determine and compare the Barr bodies present in exfoliated buccal epithelial cells in males, females and transgender populations using light and fluorescence microscopy. Materials and Methods: A total of 90 patients were recruited for the study. Group I consisted of 30 female patients. Group II consisted of 30 male patients and group III consisted of 30 transgender patients. The buccal mucosa was then scraped using a wooden spatula and the cells obtained were fixed in 95% ethanol. Two smears per individual were made and stained. One smear was stained with papanicolaou (PAP) stain and the other with Acridine orange and viewed under light microscopy and fluorescent microscopy, respectively. Results: When PAP stained slides were examined, the percentage of Barr-bodies in females ranged from 3% to 5% and in males it was 0% and in transgenders, it ranged from 0% to 5%. In Acridine orange stained smears, the percentage of Barr bodies in females ranged from 1% to 3% and in males it was 0% and in transgenders, it was 0%. Kruskal-Wallis test to study the relation of Barr body percentage in females, males and transgender subjects demonstrated significant differences between the groups (P < 0.001). Wilcoxon signed rank test was done for pairwise comparison, which showed that the distribution of percentage of positive cells in females are statistically significant from males and transgenders (P < 0. 001). Conclusion: Nuclear sexing using Barr bodies offers a simple yet effective method for determining the sex of transgender patients which could help them in understanding their gender identity better and diagnose any underlying chromosomal aberration.

Quantitative Proteomic Analysis of Non-Tobacco Associated Oral Squamous Cell Carcinoma Reveals Deregulation of Cytoskeletal and Apoptotic Proteins.

BACKGROUND: The exact etiology of non-tobacco associated oral squamous cell carcinoma (NT-OSCC) is still unknown. The lack of established biomarkers for oral NT-OSCC has resulted in less effective management and poor prognosis. Here, we report for the first time a panel of potential markers identified from the quantitative proteomic analysis of NT-OSCC using two-dimensional gel-electrophoresis (2D-GE) using matrix-assisted laser desorption/ionization - time of flight (MALDI-TOF) coupled with mass spectrometry (MS) and further analysis using protein analysis through evolutionary relationships (PANTHER) database. OBJECTIVE: To quantitatively analyze the proteomic profile of non-tobacco associated oral squamous cell carcinoma. METHODS: Twenty fresh tissue samples were collected from healthy controls and NT-OSCC, ten each, and were subjected to proteomic analysis. Sample quantification for the presence of protein was done using Bradford assay and bovine serum albumin was used as a standard protein to obtain the standard graph. Fractionation of protein was done using sodium dodecyl sulphate - polyacrylamide gel electrophoresis (SDS-PAGE) and they were separated based on their molecular weight. MS analysis was done and the purified peptides were analysed using MALDI-TOF. PANTHER database for functional classification and pathway analysis was done for identification of protein expression. RESULTS: Our approach of combining 2D-GE with MS identified four candidate proteins including keratin, alpha-1-antitrypsin (AAT), S100 and serpin B5 with significant differential expression in NT-OSCC as compared with healthy controls. The results showed that the levels of these proteins were significantly upregulated in NT-OSCC when compared to the healthy controls that suggests that these proteins can be used as candidate targets for NT-OSCC therapeutics. CONCLUSION: The differentially expressed proteins are found to be involved in apoptotic signalling pathways, cytoskeletal dynamics and are known to play a critical role in oral tumorigenesis. Put together, the results provide available baseline information for understanding the development and progression of NT-OSCC. These identified proteins on further validation may be used as potential biomarkers in future for early detection and predicting therapeutic outcome of patients with NT-OSCC.

Anti-fibrotic activity of licorice extract in comparison with colchicine on areca nut-induced fibroblasts: An in vitro study.

Objective: Oral submucous fibrosis (OSMF) is a debilitating chronic disease of the oral cavity with a high potential for malignant transformation. The main etiological agent attributed to the development of OSMF is the use of smokeless tobacco products like areca nut. There is no known cure for the disease. Current modalities of treatment do not provide a complete cure and often prove invasive for the patient. Herbal preparations using natural compounds and medicinal plant extracts have long since been used in India, as an acceptable, noninvasive and cost-effective method in the treatment of various diseases. Hence, the present study aims to assess the anti-fibrotic effect of licorice in comparison with colchicine on areca nut-induced fibroblasts. Materials and Methods: Extracts of areca nut, licorice and colchicine were prepared in accordance with established protocols. Human fibroblast cell lines were procured from ATCC®(PSC-201-018). Fibroblast cultures were established, and upon reaching confluence the cells were subjected to the 25 µg/ml areca nut extract for 24 h to induce fibrosis, with CCl4 used as control fibrosing agent. The areca nut and CCl4 induced cells were then subjected to varying concentration of the test antifibrotic agent, licorice extract for the periods of 24 and 48 h, with colchicine used as positive control. Total collagen quantification was done using spectrophotometry. Results: Collagen accumulation decreased with increase in the concentration of licorice extract with maximum reduction seen at 200 µg/ml. Kruskal-Wallis test was done to analyze the difference in collagen accumulation. Analysis revealed that the P < 0.05 for both periods in both the areca and CCl4 induced cell lines following the addition of licorice extract. The data were found to be statistically significant. Conclusion: The current study proves the antifibrotic efficacy of licorice in areca nut induced cell lines and hence, this agent can be used for the therapeutic management of OSMF.

Analysis of Metabolic Regulators PGC1-α and PGC1-ß in Oral Squamous Cell Carcinoma with and without Hyperglycemia.

AIM: To assess the expression of PGC1-α and PGC1-ß in Oral squamous cell carcinoma in the presence and absence of hyperglycemia. MATERIALS AND METHODS: Fresh tissue samples were collected from 14 well differentiated OSCC patients with hyperglycemia, 14 OSCC patients without hyperglycemia  and 14 healthy controls and subjected to quantitative real-time PCR to assess expression of PGC1-α and PGC1-ß. The relative gene expression of PGC1-α and PGC1-ß was calculated using the double delta Ct method. A two-fold difference was defined as over or under- expression.  To further evaluate clinicopathological association, Independent t-test was employed. RESULTS: The expression of both PGC1-α and PGC1-ß were increased in OSCC patients when compared to healthy controls and similar findings were observed on calculating the fold change healthy controls and OSCC study groups. On assessing the expression of target genes within study groups, they did not present with significant fold change and the hyperglycemic status of the individual did not contribute to the expression of the target genes as P value obtained for PGC1-α and PGC1-ß were >0.05. CONCLUSION: The hyperglycemic status of the individual does not influence the expression of PGC1-α and PGC1-ß in OSCC tissues and the cause for over-expression of the study targets in OSCC tissues must be further evaluated  to assess their potential as possible candidates for targeted therapy in OSCC patients.

Salivary Metabolomics for Oral Cancer Detection: A Narrative Review.

The development of low- or non-invasive screening tests for cancer is crucial for early detection. Saliva is an ideal biofluid containing informative components for monitoring oral and systemic diseases. Metabolomics has frequently been used to identify and quantify numerous metabolites in saliva samples, serving as novel biomarkers associated with various conditions, including cancers. This review summarizes the recent applications of salivary metabolomics in biomarker discovery in oral cancers. We discussed the prevalence, epidemiologic characteristics, and risk factors of oral cancers, as well as the currently available screening programs, in India and Japan. These data imply that the development of biomarkers by itself is inadequate in cancer detection. The use of current diagnostic methods and new technologies is necessary for efficient salivary metabolomics analysis. We also discuss the gap between biomarker discovery and nationwide screening for the early detection of oral cancer and its prevention.

Diagnosing oral lesions in immunocompromised individuals: A case report with a review of literature.

Oral lesions are often the first tell-tale sign for human immunodeficiency virus infections (HIV). Numerous oral lesions have been associated with HIV infections, some lesions such as candidiasis being more common than others. Regular oral screening can aid in identifying such lesions allowing for the early diagnosis of HIV and help in monitoring the progression of HIV in such individuals. We report a case of a family who manifested with oral lesions consistent with HIV. A review of literature on diagnosing immunocompromised individuals in clinical practice has also been summarized.

Phyto-intruders in oral tissues: A polarized light microscopic study.

OBJECTIVES: Foreign body reactions are common in the oral cavity due to its proximity to the external environment. Rarely, foreign body of plant origin may be encountered in the histopathological sections making the diagnosis problematic. The aim of the present study was to analyze the histological features of various products of plant origin emphasizing on the pathogenesis of tissue reaction occurring in response to their implantation. METHODS: This observational study included various plant products or phytoproducts commonly consumed in South Indian diet such as rice, curry leaves, coriander leaves, spinach leaves, coconut, green chilli, onion, French beans, urad dal, lentil beans, mustard seed, ginger, and garlic. Formalin-fixed specimens were routinely processed and stained with hematoxylin and eosin. The slides were viewed under light microscope and polarized microscope and evaluated by two oral pathologists. RESULTS: Each of the phytoproducts had distinctive histological appearance and exhibited positive birefringence. Phytoproducts such as rice, curry leaves, French bean, onion, and green chilli resembled pathological structures such as calcifications, ghost cells, clear cells, atypical adipocytes, and fungal hyphae, respectively. CONCLUSIONS: Plant products appear as puzzling structures in histological section posing difficulties to the pathologist. Recognition of these structures as foreign body based on their histological appearance is inevitable and their identification may avoid unnecessary delay in treatment planning. The current study serves as an atlas for the histology of extraneous material study and also as a reference for the pathologists whenever mystifying structures are encountered.

Alpha-L-fucosidase levels in patients with oral submucous fibrosis and controls: A comparative study.

Background: Oral submucous fibrosis (OSMF) in recent times has been recognized as a potentially malignant disorder (PMD) with an increased risk of developing oral squamous cell carcinoma with malignant transformation rates that vary from 0.6% to 36%. Alpha-L-fucosidase (AFU) is a lysosomal enzyme that is involved in maintaining the homeostasis of fucose metabolism. In benign and malignant tumors, the cells modulate their surface by increasing fucosylation leading to uncontrolled growth. Aims and objectives: This study was designed to estimate the levels of salivary and serum AFU in patients with OSMF and healthy controls and also to evaluate the clinical utility of salivary AFU levels over serum. Materials and Methods: Saliva and blood samples were collected from twenty participants in both the groups (OSMF and healthy controls). Serum and salivary alpha-L-fucosidase levels were measured by enzyme-linked immunosorbent assay. The data were subjected to appropriate statistical analysis. Results: We found a significant increase in alpha-L-fucosidase level in OSMF compared with healthy subjects. Pearson's correlation showed salivary alpha-L-fucosidase level to have superior sensitivity in detecting OSMF compared with serum alpha-L-fucosidase. Conclusion: The outcome of this study suggests that salivary alpha-L-fucosidase can be utilized as a biomarker in early detection of oral precancer and cancer.