Immunohistochemical Expression Of Ezrin In Oral Potentially Malignant Disorders-A Descriptive Study.

INTRODUCTION: Ezrin, also known as cytovillin, is a member of the ERM family of protein. Ezrin cross-links actin filament with the plasma membrane. They are involved in the formation of microvilli, cell-cell adhesion, maintenance of cell shape, cell motility, and membrane trafficking. Recent analysis reveals their involvement in signaling pathways. Ezrin is highly expressed in several types of human cancers, and correlation between its immunoreactivity and histopathological data as well as the patient outcome has previously been studied. OBJECTIVE: The objective of the study was to analyze the immunohistochemical expression pattern of ezrin in oral potentially malignant disorders (OPMDs), namely, oral submucous fibrosis (OSMF) with different grades and clinically leucoplakia (hyperkeratosis with various degree of dysplasia) and its use as a predictive marker for malignant transformation. SUBJECTS AND METHODS: Sample size n = 43, histopathologically confirmed cases of OPMDs (13 cases of OSMF with different grades and 30 cases of clinically leukoplakia) were retrieved from the Department of Oral and Maxillofacial Pathology. Immunohistochemistry was done using anti-ezrin antibody, and the expression was graded in terms of proportion and intensity. RESULTS: There was a significant expression of ezrin in OPMDs, and its cytoplasmic shift can be used as a predictive marker for malignant transformation. CONCLUSION: The findings of the current study revealed that the expression of ezrin in OPMDs may be related to the progression of the disease.

Expression of CD 68, CD 45 and human leukocyte antigen-DR in central and peripheral giant cell granuloma, giant cell tumor of long bones, and tuberculous granuloma: An immunohistochemical study.

BACKGROUND: Multinucleated giant cells (MNCs) form an integral part of numerous bone and soft tissue tumors, tumor-like lesions and are often associated with granulomas of immunological and nonimmunological origin. The presence of various types of giant cells depends on the lesions in which they are present which are difficult to be diagnosed under routine histological techniques. Immunohistochemistry can be used for a better diagnosis and understanding of the origin of various giant cells using various markers of immune response like human leukocyte antigen-DR (HLA-DR) and those expressed on monocytes and macrophages like CD 68 and leukocyte common antigen (LCA). MATERIALS AND METHODS: The study group consisted of 10 cases of giant cell tumor (GCT) of long bones, tuberculous granuloma, and giant cell granuloma to evaluate and analyze the expression pattern of LCA, CD 68, and HLA-DR in various giant cell lesions. RESULTS: Strong expression of CD 68 was observed in 80% of the lesions, strong and moderate expression of CD 45 observed in 70% of the lesions among and within the groups. In contrast, HLA-DR demonstrated negative expression in 80% of cases except for tuberculous granuloma where all the 10 cases showed moderate to strong immunoreactivity. CONCLUSION: CD 68 and CD 45 expression was found in central giant cell granuloma, peripheral giant cell granuloma and GCT, suggesting the origin from mononuclear phagocyte system and considering their clinical behavior of osteoclast type. High expressivity of HLA-DR in tuberculous granulomas which is an essential factor for presentation of the microbial antigen to CD 4 helper cells thus reassuring the fact that they are up-regulated in response to infection.

In vitro analysis of particle penetration of smokeless tobacco forms using egg shell membrane as a substrate.

AIMS: The aim of the following study is to determine the particle sizes of smokeless tobacco forms and thereby evaluate the degree of diffusion of the products using an egg shell membrane as a natural substrate by scanning electron microscopy (SEM). SUBJECTS AND METHODS: The particle size of smokeless tobacco forms namely mawa, gutka, khaini, and tobacco leaves was determined by image analysis and the products were subjected on an egg shell membrane subjected to artificial saliva along with constant grinding force. The processed egg shell membrane was then examined by SEM to evaluate the morphology and the degree of diffusion of these particles. RESULTS: The morphometric image analysis revealed khaini to be smallest in size followed by mawa, gutka, and tobacco leaves. The control group (egg shell membrane not subjected to any products) under SEM demonstrated intricately woven collagen meshwork, which was regular in length, branched and anastamosed with each other. Khaini exposed membrane demonstrated maximum particle diffusion with disruption of collagen meshwork. Mawa exposed membranes demonstrated minimal particle penetration which were adherent to the collagen meshwork which was irregular with increased interfibrillar space. The egg shell membrane exposed to gutka demonstrated particle penetration at higher magnification with more or less a regular collagen meshwork. The tobacco leaf exposed egg shell membrane demonstrated minimal blebs on the meshwork surface. CONCLUSIONS: The degree of diffusion gradually increased with a decrease in the particle size establishing an inverse relationship.

Antioxidant and pro-oxidant activity of Vitamin C in oral environment.

CONTEXT: Antioxidant properties and Vitamin C. BACKGROUND: Vitamin C is a naturally occurring organic compound and a potent antioxidant preventing oxidative damage to lipids and other macromolecules. It can also exhibit bimodal activity as a pro-oxidant at a higher concentration. Vitamin C has a switch over role from being an antioxidant in physiologic conditions to a pro-oxidant under pathologic conditions. A systematic review of this role would help to elucidate whether it is an antioxidant or a pro-oxidant in the oral environment. OBJECTIVE: To review studies reported in the literature elucidating the activity of Vitamin C and determine whether it is an antioxidant or a pro-oxidant. MATERIALS AND METHODS: Articles were searched in PubMed, MEDLINE using appropriate key words like "Vitamin C," "antioxidant activity," "pro-oxidant activity," "oral health" "oral disease." Hand search of journals was also performed. Articles were reviewed and analyzed. RESULTS: Search strategy reviewed 10 relevant articles which studied the dual role of Vitamin C. 65% of authors analyzed antioxidant action of ascorbic acid compared to 35% of the pro-oxidant potential. Vitamin C acts as an antioxidant and a pro-oxidant by a plethora of mechanisms. Factors determining its bimodal activity were studied, and the frequencies of their occurrence in the literature were depicted in percentage. CONCLUSION: The data validates the role of Vitamin C as an antioxidant under physiologic conditions exhibiting a cross over role as a pro-oxidant in pathological conditions. Further studies are required to substantiate its pro-oxidant activity to draw concrete conclusions.

Detection of species diversity in oral candida colonization and anti-fungal susceptibility among non-oral habit adult diabetic patients.

BACKGROUND AND OBJECTIVES: Patients with diabetes mellitus are prone to secondary infections. In this study we aim to determine the prevalence of one such secondary infection (oral Candida colonization) and evaluate the influence of local and systemic factors on the oral candidal colonization in patients with diabetes mellitus. MATERIALS AND METHODS: Forty non-insulin-dependent diabetic patients and 40 healthy individuals were included in this study. Samples were collected by using the oral rinse method. The candidal species were isolated and identified through phenotypic methods. An in vitro antifungal susceptibility profile was evaluated. Glycemic control, as determined by the glycosylated hemoglobin concentrations (HbA1c) of the study subjects, was correlated with the candidal colonization. RESULTS: Patients with diabetes showed a significantly higher prevalence of candidal colonization. The rate of carriage and density (P = 0.001) was higher. Candida albicans was the most predominantly isolated species, however, C. dubliniensis, C. tropicalis, and C. parapsilosis were also observed. Variable resistance toward the antifungal drugs (amphotericin B and fluconazole) was observed in the Candida isolated from diabetics, but not from healthy patients. Interestingly, a positive correlation was observed between glycemic control and candidal colonization. CONCLUSION: Diabetic patients had a higher candidal carriage rate, with a variety of candidal strains, which significantly varied in their resistance to routinely used anti-fungal agents. Interestingly the higher oral candidal colonization in diabetic patients is related to local and systemic factors, independent of their oral habits.

Metastasizing Ameloblastoma - a perennial pathological enigma? Report of a case and review of literature.

The Ameloblastoma is a slow growing locally invasive odontogenic epithelial neoplasm with a high recurrence rate and a low tendency to metastasize. Metastasis in Ameloblastoma was first described by Simmons and Emura in the 1920s. Slootweg and Muller proposed the term Malignant Ameloblastoma to describe a well-differentiated ameloblastoma that metastasizes but maintains the characteristic cytologic features of the original tumour and the term Ameloblastic Carcinoma to an ameloblastoma with malignant cytological features. About 2% of ameloblastomas undergo metastasis. So far there have only been two cases of Metastasizing Ameloblastoma reported from the Indian Subcontinent. We present the case of a 22-year-old male Indian patient, who presented with a diffuse swelling in the left posterior mandible. Radiographs revealed a multilocular radiolucency in the left mandible. On histopathological examination, the lesion was diagnosed as follicular ameloblastoma. Four years later the patient presented with a swelling in the left submandibular region. Histological examination revealed metastatic ameloblastoma within the cervical lymph node.

Promoter hypermethylation patterns of P16, DAPK and MGMT in oral squamous cell carcinoma: a systematic review and meta-analysis.

BACKGROUND: Oral squamous cell carcinoma (OSCC) is a common cancer world-wide that is highly lethal due to its recurrence and metastasis. Methylation is a common epigenetic mechanism that leads to gene silencing in tumors and could be a useful biomarker in OSCC. The prevalence of P16, death-associated protein kinase (DAPK) and O6-methylguanine-DNA-methyltransferase (MGMT) promoter hypermethylation in OSCC has been evaluated for several years while the results remain controversial. OBJECTIVE: The aim of this systematic review is to critically analyze and perform a meta-analysis on the various studies in the literature that have reported the promoter hypermethylation of P16, DAPK and MGMT genes in OSCC. SEARCH STRATEGY: Articles were searched and selected through PubMed. Hand search from the relevant journals was also performed. Articles were reviewed and analyzed. RESULTS: The estimated prevalence of P16 methylation was 43%, DAPK methylation was 39.7% and MGMT methylation was 39.8%. Heterogeneity in methylation prevalences and correlations with the clinical outcomes of the disease prevailed in various studies. CONCLUSION: We can conclude from our systematic review that a higher prevalence of methylation of P16, DAPK and MGMT occur in OSCC. Further studies are required to substantiate the role of methylation of P16, DAPK and MGMT as a marker in OSCC.

Amino Acid profile in oral submucous fibrosis: a high performance liquid chromatography (HPLC) study.

BACKGROUND: Collagen is a significant structural protein, the integrity of which is essential to be maintained for proper homeostasis. Oral submucous fibrosis (OSMF), being a collagen metabolic disorder, may be subject to changes in amino acid profiling. AIM: The present study was attempted to evaluate the amino acid profile to assess its feasibility as a biological marker in OSMF. MATERIALS AND METHODS: The study group comprised of 13 patents with OSMF and the normal group comprised of 13 normal patients without associated habits or systemic disorders. Venous blood was collected from the antecubital vein, plasma was separated and the plasma was then subjected to high profile liquid chromatographic analysis. RESULTS: The assay levels of threonine, alanine and tyrosine did not yield any significant results. The decreased assay levels of valine, Isoleucine and the increased assay level of methionine and glycine observed in group II yielded significant results in correlation with the control group. The decreased assay level seen in phenylalanine in group II and III in correlation with group IV is statistically significant. CONCLUSION: A few amino acids have been identified which can be used as biological markers for the severity of the disease such as valine, methionine and phenyl alanine. Large scale studies are required to elucidate the potential of these biological markers.

Immunohistochemical evaluation of oral epithelial dysplasia using cyclin-D1, p27 and p63 expression as predictors of malignant transformation.

OBJECTIVE: To evaluate the degree of expression of cyclin-D1, p27 and p63 in mild, moderate and severe dysplasia using immunohistochemical evaluation in order to illustrate their prognostic value and attempt to propose a molecular grading system for oral epithelial dysplasia. MATERIALS AND METHODS: The analysis included thirty cases of mild, moderate and severe dysplasia from Department of Oral and Maxillofacial Pathology, Saveetha Dental College, Chennai after a critical review of the Hematoxylin and Eosin (H and E) stained sections. They were subjected to immunohistochemical evaluation using the markers cyclin-D1, p27 and p63. The assessment of the expression based on staining intensity and distribution of immunohistochemical staining of the various markers was analyzed followed by statistical analysis. RESULTS: A highly significant increase in the expression of cyclin-D1 (P < 0.000) and p63 (P < 0.001) and a moderately significant decrease in the expression of p27 (P < 0.012) with the increasing severity of dysplasia was observed in our study. CONCLUSIONS: The result of our research affirms the fact that the increase in the expression of markers of cell cycle regulators such as cyclin D1, decrease in the expression of cell cycle inhibitors like p27 and increased expression of p63 in parallel with the increasing severity of dysplasia, emphasizes the use of immunohistochemical markers cyclin D1, p27 and p63 as prognostic markers for better understanding the behaviour of these potentially malignant disorders aiming towards proposing a molecular grading system for oral epithelial dysplasia to enable timely management prior to their possible malignant transformation.

Immunohistochemical profiling of Ameloblastomas using cytokeratin, vimentin, smooth muscle actin, CD34 and S100.

BACKGROUND: Ameloblastoma is characterized as a slow growing, non-metastatic and a locally invasive tumor with a high risk of recurrence. Immunohistochemical evaluation of ameloblastomas using epithelial and connective tissue specific markers help in studying the histogenesis and assessing the biological behavior. The aim of the study was to study the expression patterns of cytokeratin, vimentin, smooth muscle actin (SMA), S100 and CD34 in ameloblastomas. MATERIALS AND METHODS: The material for the study consisted of 24 cases of ameloblastomas. The excised specimens were grossed and bits were taken from different areas of the specimen. Based on the histopathology, the cases were classified into different types and stained for immunohistochemistry. RESULTS: The cases showed strong positivity to cytokeratin, vimentin, moderate positivity for SMA and S100. Five cases were also moderately positive for CD34 in blood vessels. CONCLUSION: The results and hypothesis achieved from the study, proved to be consistent, not only augmenting the already existing hypothesis but also imparting new concepts of hypothesis.

Epithelial maturation pattern of dysplastic epithelium and normal oral epithelium exposed to tobacco and alcohol: a scanning electron microscopic study.

BACKGROUND: The detection of oral cancer at an early stage is an optimal strategy and is the most effective approach for preventing further progression. The rationale of the study was to evaluate the epithelial maturation pattern in oral mucosa exposed to tobacco/alcohol and on dysplastic oral mucosa using the scanning electron microscope. METHODS: Fifteen subjects were selected based on clinical examination and divided into three groups: group 1-patients with apparently normal oral mucosa; group 2-patients with oral mucosa exposed to tobacco/alcohol; group 3-patients with clinical diagnosis of leukoplakia. An incisional biopsy was performed from the buccal mucosa. One part of the specimen was prepared for light microscopy and the other part was prepared for scanning electron microscopy. RESULTS: Light microscopy revealed nonkeratinized stratified squamous epithelium in group 1, while group 2 demonstrated hyperparakeratinized stratified squamous epithelium with mild cytological atypia, and group 3 showed architectural and cytological changes. Scanning electron microscopy demonstrated flat-surfaced cells with equidistant parallel microridges in group 1, while group 2 showed irregular and widened microridges with numerous pits and absence of honeycomb pattern. Group 3 showed irregularly arranged broad and swollen cells with numerous pits and irregular microvilli projecting over the surface. CONCLUSION: The present study establishes the relationship of the surface abnormalities to the tendency of the cells to become malignant and thus serves as a tool in early detection of squamous cell carcinoma. It also emphasizes the need of routine follow-up in these high-risk patients for progression of carcinoma.

Ascorbic acid and its pro-oxidant activity as a therapy for tumours of oral cavity -- a systematic review.

BACKGROUND: Ascorbic acid or Vitamin C is a potent dietary antioxidant with a double faced character, in that it exhibits a pro-oxidant activity arising from its routine antioxidant property that generates reactive free radicals, which induce cytotoxic effects at pharmacologic concentrations. A systematic review of this effect of ascorbic acid in the oral tumours and normal oral tissues would clearly elucidate the merits or demerits of employing vitamin C in treating the same. OBJECTIVE: The aim of our systematic review is to critically review the studies reported in literature that have studied the pro-oxidant activity of ascorbic acid as a therapeutic option for treatment of oral neoplasms and its effects on normal oral cells. METHODS: Articles were searched in PUBMED, MEDLINE using appropriate key words like "ascorbic acid", "pro-oxidant activity", "treatment", "oral neoplasms". Hand search of Journals was also performed. Articles were reviewed and analysed. RESULTS: The search strategy included 17 potentially relevant articles for review of which, 12 were in vitro studies; 3 were in vivo animal studies; 1 was in vivo human study and 1 was ex vivo human study. The optimum concentration of ascorbic acid used to produce potential pro-oxidant associated cytotoxic effects was found to be 3-5mM in vitro, 0.88-5mM in vivo animals, 0.5-2mM ex vivo in humans, and the corresponding effects are induction of apoptosis (caspase activation), necrosis, free radical formation, H2O2 generation, and DNA fragmentation. In contrast, the same pro-oxidant concentrations had no effect on the normal cells. CONCLUSION: The results of our systematic review show that the pro-oxidant activity of pharmacologic ascorbic acid is a part of its dose-dependent bimodal activity and is a result of the proposed Fenton mechanism. In vitro, animal and ex vivo studies of pharmacologic ascorbic acid (AA) have yielded meritorious results proving vitamin C as an effective cytotoxic agent against oral neoplastic cells with potentially no harming effects on normal cells. However, a shortage of clinical trials and in vivo human studies pertaining to evaluation of anti-tumour activity of vitamin C in tumours of oral cavity remains a lacuna in concluding ascorbic acid as a beneficial therapeutic option in treatment of oral neoplasms.

Genetic alterations in syndromes with oral manifestations.

Ever since Gregor Johan Mendel proposed the law of inheritance, genetics has transcended the field of health and has entered all walks of life in its application. Thus, the gene is the pivoting factor for all happenings revolving around it. Knowledge of gene mapping in various diseases would be a valuable tool in prenatally diagnosing the condition and averting the future disability and stigma for the posterity. This article includes an array of genetically determined conditions in patients seen at our college out-patient department with complete manifestation, partial manifestation and array of manifestations not fitting into a particular syndrome.

Amino acid profile of saliva from patients with oral squamous cell carcinoma using high performance liquid chromatography.

Oral cancer is the leading cause of death worldwide and it is the eighth most common cause of cancer death. Cancer cells utilize more glucose and amino acids than their benign counterparts. Diagnosis of disease via the analysis of saliva is potentially valuable, as the collection of fluid is associated with fewer compliance problems than the collection of blood. Hence, the present study was undertaken to evaluate the comprehensive amino acid profiling of saliva by high performance liquid chromatography (HPLC). The study group comprised 16 subjects, of whom eight were classified as having well-differentiated oral squamous (OSCC) cell carcinoma (Group I) and eight were classified as having moderately differentiated oral squamous cell carcinoma (Group II). Eight healthy individuals comprised the control group (Group III). The results showed increased salivary levels of all the amino acids in both groups of OSCC patients (Groups I and II) when compared with healthy controls (Group III). Hence, our study showed higher levels of all amino acids in the saliva of OSCC patients than in the saliva of healthy controls. The increased levels may serve as a "diagnostic and prognostic marker" for oral squamous cell carcinoma and for further detection of metastatic spread.

Micronuclei and other nuclear anomalies in buccal mucosa following exposure to X-ray radiation.

OBJECTIVE: To evaluate DNA damage and cellular death in exfoliated buccal mucosa cells from healthy individuals following dental X-ray exposure, to compare effects of different types of radiographic techniques using the buccal cytome assay and to standardize the staining procedure. STUDY DESIGN: This study comprised 90 patients, categorized into 3 groups subjected to intraoral periapical radiography, conventional orthopantomogram and digital orthopantomogram X-ray exposure. Exfoliated oral mucosa cells were collected immediately before the X-ray and 10 days later and stained using DNA-specific stains and nonspecific DNA stains. RESULTS: The results indicated no statistically significant differences in micronucleated cells before and after dental X-ray exposure. On the other hand, X-ray exposure did increase other nuclear alterations closely related to cytotoxicity, such as karyorrhexis, pyknosis and karyolysis. CONCLUSION: These data indicate that dental radiography may not induce chromosomal damage, but it is able to promote cytotoxicity.

Lichen sclerosus of the oral cavity. Case report and review of literature.

BACKGROUND: Lichen sclerosus is a chronic muco-cutaneous inflammatory disorder of uncertain etiology. The prevalence of lichen sclerosus affecting only the oral mucosa is extremely rare and only 26 cases with histologically verified oral lichen sclerosus have been reported. MAIN OBSERVATIONS: A 60-year-old man was referred to our hospital for complaints of white lesions in the oral cavity, which was otherwise asymptomatic and did not have any cutaneous or anogenital lesions. Clinical examination revealed white patches with reddish areas on the buccal mucosa bilaterally and on the maxillary posterior gingiva. Microscopic analysis of the lesion showed atrophied epithelium with basal cell degeneration, hyalinized connective tissue stroma with minimal chronic inflammatory cell infiltrate. Verhoeff's staining revealed scantiness of elastic fibers in the connective tissue stroma. On the basis of these histological findings, the final diagnosis was given as Lichen Sclerosus. LS is rare in the oral cavity, particularly in the absence of simultaneous cutaneous and anogenital lesions. CONCLUSIONS: Only 7 cases of oral LS have been reported involving the gingiva. To our knowledge, this is the eigth case to be reported with gingival involvement and the first case to be reported with bilateral involvement of buccal mucosa and gingiva.

Glandular odontogenic cyst: report of two cases and review of literature.

Glandular odontogenic cyst (GOC) is an uncommon jaw bone cyst of odontogenic origin described in 1987 by Gardner et al. It is a cyst having an unpredictable and potentially aggressive behaviour. It also has the propensity to grow to a large size and tendency to recur with only 111 cases having been reported thus far. The first case occurred in a 42-year-old female and presented as a localized swelling extending from 19 to 29 regions. There was a history of traumatic injury at the site. There was evidence of bicortical expansion and radiographs revealed a multilocular radiolucency. The second case occurred in a 21-year-old male, as a large swelling in the mandible and radiograph revealed radiolucency in the region. On histopathological examination, these lesions were diagnosed as GOC. It was concluded that, two cases submitted by us correlate with the existing literature that GOC's affect more commonly in the middle age group, having predilection for mandible and that trauma could be a precipitating factor for its occurrence. The increased recurrence rates can be due to its intrinsic biological behavior, multilocularity of the cyst, and incomplete removal of the lining following conservative treatment.

Epidermoid cysts: an exclusive palatal presentation and a case series.

BACKGROUND: Epidermoid cysts (EC) can occur anywhere in the body. In the head and neck region, they more often present in the midline between the submental region and the supra sternal notch. EC in the oral cavity are extremely rare and present as benign, slow growing lesions. METHODS: Records from College of Dental Surgery, Saveetha University, from 2002 to 2006 were searched for cases coded as EC. The study included 13 cases within the oral and maxillofacial region. RESULTS: Of the 13 cases, 11 occurred in male patients and 2 in female patients. Specific anatomic locations included posterior auricular region (n = 2), forehead region (n = 2), lateral side of the face (n = 3), chin (n = 1), gingiva (n = 3), intraosseous within the anterior maxilla (n = 1), and hard palate (n = 1). CONCLUSION: Besides the previously reported locations, we present the first case occurring in the hard palate, measuring 5 x 3 cm in size along with melanin pigmentation.