Diminishing reactive adipogenesis leads to disease progression of oral submucous fibrosis.

Oral fibroblasts, similar to dermal fibroblasts, have the potential to resist the local insults like trauma to the oral mucosa by differentiating into adipocytes and secreting antimicrobial peptide cathelicidin (Camp) and this physiologic process in known as reactive adipogenesis. We hypothesize that in oral submucous fibrosis (OSF), due to constant secretion and up-streaming of transforming growth factor-beta (TGF- ß), oral fibroblast lose their adipogenic differentiation potential and Camp production, which leads to progressive fibrosis in OSF. The implication of this hypothesis could open some promising vistas on still unexplored innate immune systems harboured by oral mucosa. Restoring and maintaining the adipogenic and protective potential of oral fibroblasts by inhibiting TGF- ß receptors could hinder the disease progression of OSF.

Proteomic profiling of various human dental stem cells - a systematic review.

BACKGROUND: The proteomic signature or profile best describes the functional component of a cell during its routine metabolic and survival activities. Additional complexity in differentiation and maturation is observed in stem/progenitor cells. The role of functional proteins at the cellular level has long been attributed to anatomical niches, and stem cells do not deflect from this attribution. Human dental stem cells (hDSCs), on the whole, are a combination of mesenchymal and epithelial coordinates observed throughout craniofacial bones to pulp. AIM: To specify the proteomic profile and compare each type of hDSC with other mesenchymal stem cells (MSCs) of various niches. Furthermore, we analyzed the characteristics of the microenvironment and preconditioning changes associated with the proteomic profile of hDSCs and their influence on committed lineage differentiation. METHODS: Literature searches were performed in PubMed, EMBASE, Scopus, and Web of Science databases, from January 1990 to December 2018. An extra inquiry of the grey literature was completed on Google Scholar, ProQuest, and OpenGrey. Relevant MeSH terms (PubMed) and keywords related to dental stem cells were used independently and in combination. RESULTS: The initial search resulted in 134 articles. Of the 134 full-texts assessed, 96 articles were excluded and 38 articles that met the eligibility criteria were reviewed. The overall assessment of hDSCs and other MSCs suggests that differences in the proteomic profile can be due to stem cellular complexity acquired from varied tissue sources during embryonic development. However, our comparison of the proteomic profile suffered inconsistencies due to the heterogeneity of various hDSCs. We believe that the existence of a heterogeneous population of stem cells at a given niche determines the modalities of regeneration or tissue repair. Added prominences to the differences present between various hDSCs have been reasoned out. CONCLUSION: Systematic review on proteomic studies of various hDSCs are promising as an eye-opener for revisiting the proteomic profile and in-depth analysis to elucidate more refined mechanisms of hDSC functionalities.

Comparison of the Efficacy of Sediment Cytology over Oral Brush Cytology in Oral Leukoplakia.

INTRODUCTION: A method called sediment cytology includes the investigation of smears arranged from the sediment of the biopsy specimen fixatives. The sediment from this fixative is used to prepare smears and provides a potentially rich source for cytological material. Investigation of the fixative sediment and understanding of the cytological picture with pertinent clinical and radiological information permits diagnosis in a few hours. AIM: To evaluate the diagnostic efficacy of sediment cytology and oral brush cytology compared with histopathological findings in oral leukoplakia (OL) cases. METHODS: Cytological smears were obtained from 30 clinically diagnosed OL lesions using 2 techniques: oral cytobrush and 10% formalin fixative sedimentation. Both smears were stained with Papanicolaou. Cytological smear evaluation was conducted with respect to cellularity, cell distribution, cellular clumping, and the presence of blood, debris, inflammatory cells, and microbial colonies. The cytopathological scores for all cases were compared between sediment and brush cytology and correlated with the histopathological diagnosis. For statistical analysis, the κ test and the Wilcoxon matched-pair test were used. RESULTS: The cytobrush technique had a sensitivity of 83.3% for OL cases histopathologically diagnosed as severe dysplasia, while the sediment cytology technique had a sensitivity of 16.6%. For moderate/mild dysplasia cases, the cytobrush technique had a sensitivity of 7.7%, whereas the sediment technique showed no diagnostic sensitivity. CONCLUSION: Based on the results from the present study, sediment cytology, unlike oral brush cytology, is not a useful screening tool for the preliminary diagnosis of potentially malignant oral lesions.

Comparative Evaluation of Immunohistochemical Expression of p16 with p16 Microsatellite Marker by PCR in Surgical Margins of Oral Squamous Cell Carcinoma.

Oral squamous cell carcinoma (OSCC) is most common oral cancer with multifactorial etiology. Surgical therapy is treatment of choice but known to have recurrence. The main reason for recurrence is associated with surgical margins which need to be tumor free. Changes at genetic level cannot be ascertained only through routine light microscopy in surgical margins, even though they are tumor free. Detection of early marker like p16 can help in predicting the risk of recurrence. Hence study aimed to detect p16 microsatellite marker (D9s1747) in surgical margins of oral squamous cell carcinoma and compare the same with p16 marker through immunohistochemistry. Total of 40 paraffin embedded tissue samples diagnosed and surgically treated cases of OSCC were included. From each sample one tumor proper and one surgical margin was obtained. From paraffin embedded tissue sample 2 sections of 4 µm thick was obtained from tumor proper and tumor margin. One section was stained with hematoxylin and eosin and other section was stained immunohistochemically using p16 antibody. DNA extraction was done for tumor proper and surgical margin tissue and PCR analysis was carried for p16 microsatellite marker (D91747). Out of 40 cases 37 cases showed positivity in tumor proper for p16 with IHC. Out of 37 cases 23 cases showed positivity for both tumor proper and surgical margin. There were 3 cases negative for tumor proper. Out of these 3 cases, 1 (33.3%) case was positive for surgical margin. Out of 40 cases 27 cases showed positivity for tumor proper with p16 microsatellite marker. Out of 27 cases 16 cases showed positivity for both tumor proper and surgical margin. There were 13 cases negative for tumor proper. However there were 8 (61.5%) cases negative which were in tumor proper but showed positivity for surgical margin. Other 5 cases were negative in both tumor proper and surgical margin. Our study reveals that surgical margins of OSCC exhibit alteration in p16 markers both by IHC and PCR techniques. p16 and p16 microsatellite marker detection in margins indicates field change. Further studies with larger sample size comparing expression with clinical and histological parameter and follow up has to be done to substantiate our findings.

Cell-free tumor DNA: Emerging reality in oral squamous cell carcinoma.

The intention of this review was to condense ongoing findings on the use of circulating DNAs from bodily fluids (blood, serum and plasma) as cancer biomarkers in patients with oral squamous cell carcinoma. Studies were collected after searching databases: PubMed and Google library. Additional search was performed through cross-check on the bibliography of selected articles. After the selection process made by two of the authors, articles which met the inclusion criteria were included in the review. Results revealed that circulating DNAs from blood, serum or plasma appear as favorable candidates as cancer biomarkers in patients suffering from oral cancer. The possibility to forecast recurrences and metastases through follow-up by quantification of candidate DNAs serve as another possible characteristic to be directed in forthcoming studies. However, methodological standardization and even sampling are required to increase the power and accuracy of results.

Detection and comparison of microsatellite marker D9S1747 with clinical stages and grades of oral squamous cell carcinoma.

BACKGROUND: One of the main characteristics of oral squamous cell carcinoma (OSCC) is genetic alteration in specific target regions. Allelic imbalance in tumor suppressor genes is the key event in OSCC which is associated with loss of heterozygosity mostly on chromosome 9p21 locus which includes p16 marker. p16 (D9S1747) is a microsatellite marker which detects early changes in OSCC. To redefine more clearly the role of D9S1747 (p16 microsatellite marker) and its expression in OSCC, the study was designed with the aim to check the detection of D9S1747 in OSCC and to compare the same with histopathological grades and tumor node metastasis staging. MATERIALS AND METHODS: Forty cases of paraffin-embedded tissue section which was histologically confirmed as OSCC and 10 cases of normal tissues were retrieved from the archives. DNA was extracted from the tissue sections and subjected for polymerase chain reaction to detect p16 microsatellite marker D9S1747. Data were analyzed using Chi-square test and Fisher's exact test. RESULTS: Twenty-seven cases (67.5%) showed p16 microsatellite marker positivity for OSCC. It was observed that 44.4%, 51.9% and 3.7% p16 microsatellite markers were positive in Stage 1, Stage 2 and Stage 4 OSCC cases, respectively. p16 microsatellite marker positivity was found in 77.8%, 22.2% and 0% for well-differentiated, moderately differentiated and poorly differentiated OSCC cases, respectively. CONCLUSION: The observations of the present study revealed D9S1747 marker as an early event in OSCC, and this can be used as a prognostic marker.

Detection of antibodies against Aggregatibacter actinomycetemcomitans in serum and saliva through ELISA in periodontally healthy individuals and individuals with chronic periodontitis.

BACKGROUND: Periodontitis is a persistent polymicrobial infection, which leads to chronic inflammation in the tooth supporting tissues. Aggregatibacter actinomycetemcomitans are normal commensals of oral cavity but are low in number in periodontally healthy subjects. They are one of the major pathogens aetiologically linked to periodontal disease. Plasma and salivary antibody measurement may be useful to support diagnosis, disease activity, classification and prognosis of periodontitis. The aim of the present study was to investigate the association between the serum and salivary antibody levels to A. actinomycetemcomitans and therefore, to find whether this association was varying in different grades of periodontitis. METHOD: Total of 50 periodontally healthy and 50 chronic periodontitis subjects (35-65 years) of both sexes were included for the study. 2 ml of un-stimulated saliva and 5 ml of venous blood was collected under sterile conditions. The detection of antibodies against A. actinomycetemcomitans in periodontally healthy individuals and individuals with chronic periodontitis was performed using indirect ELISA. RESULTS: Results showed serum IgG, IgA mean levels against A. actinomycetemcomitans were higher in chronic periodontitis subjects compared to mean levels in periodontally healthy subjects. Similarly, salivary IgG, IgA levels were also raised in chronic periodontitis patients as compared in healthy subjects. Also the mean levels of serum IgG and salivary IgA were increased as the severity of disease increased. CONCLUSION: Antibody titer using saliva and serum could be useful tool for screening of patients with chronic periodontitis. Further, monitoring the various phases of treatment outcome using saliva could be a useful, non-invasive, prognostic indicator.

Comparison of palatal rugae pattern among Indian and Tibetan population.

INTRODUCTION: Palatoscopy is the study of palatal rugae pattern to establish the identity of a person. The palatal rugae are permanent and unique to each person and can establish identity through discrimination (via casts, tracings, or digitized rugae patterns). In addition, rugae pattern may be specific to racial groups facilitating population identification (which may require postdisasters). Hence, they can be used in postmortem identification provided an antemortem record exists. AIM: To determine the palatal rugae pattern and to assess the predominant palatal rugae pattern in Indian and Tibetan (in Mundgod Taluka, Karnataka) populations. MATERIALS AND METHODS: The impressions of the maxillary arch were made for a total of one hundred adults comprising fifty Indian and fifty Tibetan populations aged between 20 and 40 years, and the dental cast was made using dental stone. The rugae were highlighted by a sharp graphite pencil on the cast under adequate light and a magnification lens. Rugae patterns were assessed using Thomas and Kotze and Kapali et al. classification. RESULTS: Total number of palatal rugae in Indian population (461) was more than Tibetan population (351). Moreover, Indian population showed predominantly wavy (43.60%) rugae pattern, whereas Tibetan showed curved (38.2%) rugae pattern. CONCLUSION: This suggests that there is a difference in the rugae pattern between Indian and Tibetan populations. Hence, palatal rugae pattern can be used as one of the methods in determining the ethnicity.

Do the presence of Barr bodies in male jail inmates indicates criminality: A pilot study.

BACKGROUND: Cytogenetic studies from past decades have shown that interphase cells of female cats contain a densely stained chromatin mass in their nuclei called as Barr bodies (BBs) named after the scientist Murray Barr. BBs are unique chromatin structures formed due to the condensation of the X-chromosome. Many psychopathic disorders originate from defective genes including the multiple X syndromes. Males with extra X-chromosome generally present with severe personality disorder. The present study was conducted to determine the presence of extra X-chromosome in male jail inmates through the detection of BB in peripheral blood and buccal smear. MATERIALS AND METHODS: Study included 100 male subjects (fifty jail inmates and fifty controls), after obtaining the consent, peripheral blood smears (PBS) and buccal smears (BS) were prepared and stained using Leishman's and cresyl violet stain respectively. One hundred neutrophils in PBS and epithelial cells in BS were screened for detection of the BB; accumulated data were tabulated and statistically analyzed using t-test and Chi-square test. RESULTS: 60% of cases in PBS and 36% in BS showed positivity for the presence of BB in jail inmates as compared to 14% of cases in PBS and none in BS were positive for BB in controls. CONCLUSION: Presence of BB in male suggests increased likelihood of criminal tendencies. Further studies are to be carried out to compare the results with karyotyping.

Immunohistochemical Expression of CD105 and TGF-ß1 in Oral Squamous Cell Carcinoma and Adjacent Apparently Normal Oral Mucosa and its Correlation With Clinicopathologic Features.

Angiogenesis in oral squamous cell carcinomas (OSCC) is essential for its growth, invasion, and metastasis. This entails a shift in the balance between proangiogenic and antiangiogenic factors. CD105 and TGF-ß1 are 2 such proangiogenic factors wherein CD105 exerts its angiogenic effect by binding to and modulating the TGF-ß1 pathway. A total of 50 resected specimens of OSCC were considered. One tissue specimen was taken from tumor proper and another specimen from adjacent apparently normal mucosa (AANM). Both tissues were immunohistochemically stained using CD105 and TGF-ß1 antibodies. The expression of each antibody was individually assessed and then compared. Pearson χ test was used for statistical comparison of expression. CD105 was significantly expressed in OSCC as compared with AANM and also correlated with increasing TNM stage. The mean microvessel density was higher in OSCC. TGF-ß1 was significantly expressed in epithelium of OSCC as compared with AANM. On comparing expression of TGF-ß1 and CD105, 79.54% of endothelial cells expressed positivity for both molecules. Both CD105 and TGF-ß1 were increased in OSCC, although based on our results CD105 alone can be used as a prognostic marker. On the basis of immunohistochemical expression of CD105 and TGF-ß1 in endothelial cells, our results demonstrate that CD105 acts as one of the receptors of TGF-ß1 on endothelial cells and induces the angiogenic pathway in OSCC.

Histomorphometric study to compare histological changes between oral squamous cell carcinoma and apparently normal adjacent oral mucosa.

Despite the advances in surgery, radiotherapy and chemotherapy the annual death for oral squamous cell carcinoma (OSCC) is rising rapidly. The carcinoma has propensity to develop in a field of cancerization. Clinically may it be apparently normal mucosa (ANM) adjacent to squamous cell carcinoma which harbours certain discrete molecular alteration which ultimately reflects in cellular morphology. Hence the aim of the study is to assess histomorphometric changes in ANM adjacent to OSCC. A prospective study was done on 30 each of histologically diagnosed cases OSCC, ANM at least 1 cm away from OSCC, and normal oral mucosa (NOM). Cellular and nuclear morphometric measurements were assessed on hematoxylin and eosin sections using image analysis software. Statistical analysis was done using analysis of variance test and Tukey's post hoc test. The present study showed significant changes in cellular and nuclear area in superficial and invasive island of OSCC compared to ANM. The basal cells of ANM showed significant decrease in cellular and nuclear areas and nuclear cytoplasmic ratio when compared to NOM. Histomorphometry definitely can differentiate OSCC form ANM and NOM. The basal cells of ANM showed significant alterations in cellular area, nuclear area and nuclear cytoplasmic area when compared to NOM suggesting change in the field and have high risk of malignant transformation. These parameters can be used as indicator of field cancerization.

Myofibroblast presence in apparently normal mucosa adjacent to oral squamous cell carcinoma associated with chronic tobacco/areca nut use: evidence for field cancerization.

OBJECTIVE: Myofibroblasts are primary cellular components of activated tumor stroma, associated with poor prognosis in oral squamous cell carcinoma (OSCC). However, their role in field cancerization has not been addressed. This study aims to evaluate the presence of myofibroblasts in patient-matched histologically normal mucosa adjacent to oral squamous cell carcinoma (HNMAOSCC) and OSCC tissues. MATERIALS AND METHODS: Fifty patient-matched tissues of OSCC and HNMAOSCC associated with chronic areca nut/tobacco use were subjected to immunohistochemistry using α-SMA for detection of myofibroblasts. Normal oral mucosa (n = 15) were stained as controls. RESULTS: The number of α-SMA stained myofibroblasts in OSCC and HNMAOSCC were significantly increased as compared to that of the normal controls (p < 0.001). Further, a significant correlation was established for the presence of myofibroblasts in the stroma of OSCC and HNMAOSCC. CONCLUSIONS: Myofibroblasts are an early stromal change in the HNMAOSCC, highlighting the possible role of myofibroblasts as likely mediators for field cancerization and their potential use as a field effect marker.

Reliability of mandibular canines as indicators for sexual dichotomy.

INTRODUCTION: Amongst the various calcified structures in the human body, teeth have gained lot of popularity in estimating the sex of an individual as they are highly resistant to destruction and decomposition. Using permanent mandibular canines many researchers have predicted a high level of accuracy in identifying the sex correctly. The purpose of our study was to gauge the effectiveness of mandibular canines in discerning sex. MATERIALS & METHODS: Fifty dental casts each of males and females were utilized for the study. Mesio-distal dimension and inter-canine distance of mandibular right and left canine was recorded using digital vernier caliper and mandibular canine index was calculated. RESULTS: The mean value of mesio-distal dimensions of right and left mandibular canine was slightly greater in males compared to females. The mandibular canine index was equal in both sexes. Inter-canine distance was marginally higher in males compared to females. Despite of higher values in males none of the parameters were statistically significant. CONCLUSION: The results herein bolster contemporary studies that mesio-distal dimensions of mandibular canines and mandibular canine index do not reflect sexual dimorphism and that its application should be discontinued in sex prediction among Indian populations. How to cite this article: Hosmani J V, Nayak R S, Kotrashetti V S, Pradeep S, Babji D. Reliability of Mandibular Canines as Indicators for Sexual Dichotomy. J Int Oral Health 2013; 5(1):1-7.

Analysis of Expression and Localization of TLR-2 by Immunofluorescent Technique in Healthy and Inflammed Oral Tissues.

INTRODUCTION: Toll-like receptors (TLRs) are an important component of immune system. Among them, TLR-2 plays a dominant role in the oral tissues in initiating inflammation in chronic periodontitis. Not many studies have been done on quantitative expression of TLR-2 by using immunofluorescent techniques (IFT) in oral tissues. In this study, the expression and localisation of TLR-2 were detected in gingival tissues of chronic periodontitis patients and healthy individuals. MATERIAL AND METHODS: Immuno Fluorescent Technique (IFT) was used for the expression and localization of TLR-2 in gingival tissue samples from 25 chronic periodontitis patients and from 25 healthy controls. Haematoxylin and Eosin staining was also done for all the samples to determine the histological characteristics of the gingival tissue samples. RESULT: Both healthy and periodontitis gingival tissues expressed TLR-2. We found that the expression level of TLR-2 was higher in all the periodontitis patients than in healthy individuals. We also found out that the expression of TLR-2 was higher in the epithelial cells than in the connective tissue cells. CONCLUSION: These data suggest a definite involvement of TLR-2 in initiating an inflammatory response in periodontal tissues. More studies are required to define the mechanisms and expression levels of TLR-2 in oral health and diseases.

Establishment of field change by expression of cytokeratins 8/18, 19, and MMP-9 in an apparently normal oral mucosa adjacent to squamous cell carcinoma: A immunohistochemical study.

BACKGROUND: Squamous cell carcinoma is the most frequent malignancy of the oral cavity. The survival rate of this malignancy has not improved from past two decades. The major factors responsible for this could be due to loco regional and distant metastatic spread. However, the other important prognostic factor is concomitant occurrence and recurrence of multiple primary carcinomas in the head and neck region, which is explained as the concept of field cancerization. The evidence to support the field change in normal mucosa of Oral squamous cell carcinoma (OSCC) through biological markers using immunohistochemistry has always been challenging. AIM: Hence, the aim of the present research is to identify changes in the expression of CK 8/18, 19, and MMP-9 to visualize field changes in the clinically normal mucosa adjacent to OSCC and compare with non neoplastic normal oral mucosa. MATERIALS AND METHODS: 20 cases of OSCC with radical resection specimens were included in the study. Lesional tissue and adjacent normal looking mucosa were taken during grossing. Ten cases of non-neoplastic normal oral mucosa are also included in the study. Markers such as CK 8/18, CK 19, and MMP-9 are used by the immunohistochemical method in this present study. RESULT AND CONCLUSION: The enhance expression of CK 8/18 (80%), CK 19 (70%), and MMP 9 (90%) in ANM was noted and furthermore in six ANM showing severe dysplasia with enhance expression of CK 8/18, CK 19, and MMP 9 in the apparently normal oral mucosa can suggest a field cancerization.