Diagnoses of 964 oral biopsies from people aged over 50 years in Karnataka State, India.

INTRODUCTION: The drastic epidemiological transition has created a huge void in evidence with regard to the health statistics of elderly population. Data are sparse on the oral mucosal conditions prevalent among the elderly Indian population. This study aimed to review the oral biopsies reported in the institute over the last 21 years and to examine the trends in geriatric oral pathologies. METHODOLOGY: A total of 964 histologically confirmed geriatric (age ranging 50-97 years) oral lesions among 4000 cases reported were retrieved from the files of Department of Oral Pathology and Microbiology of KLE VK institute, Belgaum, Karnataka commencing since 1992 till 2012. These cases were retrospectively evaluated for various parameters such as age, gender, duration, location and type of lesions. RESULTS: The geriatric oral lesions were seen to occur predominantly in men (65%), especially the sixth decade of life (47%). The most common location was the buccal mucosa (29%). The malignant neoplastic lesions accounted for the majority of the geriatric lesions (36%) followed by non-neoplastic lesions (27%), potentially malignant disorders (26%) and benign neoplasms, which formed a minor proportion of (11%). The oral squamous cell carcinoma was the single most common geriatric lesion accounted. Gender and age-wise correlation analyses among the various lesion categories showed statistical significance. CONCLUSION: The study provides valuable information about major oral pathologies in our scenario that contribute to significant morbidity and mortality among the geriatric population. It also sheds light about the diseases that need to be targeted in future Indian public health policies and initiatives for geriatric care.

Evaluation of cannibalistic cells: a novel entity in prediction of aggressive nature of oral squamous cell carcinoma.

OBJECTIVE: Cellular cannibalism is a distinctive morphologic feature exclusively seen in aggressive malignancies and is defined as a large cell enclosing a slightly smaller one within its cytoplasm. This phenomenon has been illustrated in several malignancies and is correlated well with the degree of anaplasia, invasive and metastatic potential of tumor cells. However, this marker has not been extensively studied and is often unnoticed during the routine histopathological assessment of Oral squamous cell carcinoma (OSCC). Thus, the aim of this research was to evaluate the presence of cannibalistic cells (CC) and to find if there exists any association with the aggressive nature of OSCC. MATERIALS AND METHODS: In total, 20 neck dissection cases of OSCC with follow-up data were included in the study. The cannibalistic cells were evaluated in the lesion tissues. Cellularity of cannibalism is graded asGrade I: < 5 cells, Grade II: 6-15 cells and Grade III:> 16 cells. The data was analyzed using Fischer Exact test. RESULTS: Out of 20 cases, all the cases showed presence of CC, Grade I in five cases, Grade II in eight cases and Grade III in seven cases. A statistically significant relation between advanced grade of cellular cannibalism and lymph node positive status (p≤ 0.001) was obtained. CONCLUSION: Interestingly the cases with positive lymph node metastasis demonstrated Grade 3 CC.Hence, during routine histopathological examination, the search of CC can be considered as one of the important parameters to note the aggressive nature of OSCC.

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.

Can quantifying free-circulating DNA be a diagnostic and prognostic marker in oral epithelial dysplasia and oral squamous cell carcinoma?

PURPOSE: Previous studies have reported significantly higher concentrations of serum DNA in various types of cancers. Thus the study aims to determine whether circulating free DNA (CFDNA) can aid in the diagnosis and prognosis of oral epithelial dysplasia and oral squamous cell carcinoma (OSCC). MATERIALS AND METHODS: A nonrandomized case-control study was planned where cases were derived from patients who presented to the KLE Institute of Dental Sciences, Belgaum, India, for evaluation and management of oral lesions between 2007 and 2009. In this study the predictor variable was status of the disease in the patients, and the outcome variable was CFDNA. Demographic variables included age, gender, tobacco consumption, and stage at diagnosis of cancer. Subjects with any known systemic disease, other tumors, or metastatic OSCC were excluded (CFDNA is altered in cases of tissue destruction and inflammatory diseases). The amount of CFDNA was determined through spectrophotometry (NanoDrop ND-1000 spectrophotometer; Thermo Fisher Scientific, Waltham, MA) in sampled blood and plasma. Mean and range for DNA quantification in plasma and blood were calculated in all groups and compared by use of the analysis of variance test. RESULTS: A total of 390 cases (90 potentially malignant lesions, 150 OSCC cases, and 150 post-treatment OSCC cases) and 150 control subjects were studied. No significant difference was observed in levels of CFDNA in blood between the groups. Similarly, levels of CFDNA in plasma showed no significant difference except between the OSCC and potentially malignant groups, which was probably artifactual. CONCLUSIONS: This study shows that disease progression in oral malignancy does not correlate with changes in levels of CFDNA in blood or plasma.

Cytological assessment of Barr bodies using aceto-orcein and papanicolaou stains in buccal mucosal smears and their sex estimation efficacy in an Indian sample.

OBJECTIVE: Buccal epithelial cells in saliva traces found at a crime scene can be used for sex determination by examining the presence of Barr bodies in the nucleus. Papanicolaou (PAP) and fluorescent stains, among others, have been used in the past. Aceto-orcein (AO) is a relatively unexplored staining technique for this purpose. This study aims to assess the efficacy of sex determination using AO and PAP stains for the detection of Barr bodies in buccal mucosal scrapes. STUDY DESIGN: Buccal scrapings were collected from 120 healthy individuals (60 males and 60 females). They were stained with AO and PAP. Fifty cells in each sample were analyzed for identification of Barr bodies. Samples with a presence of Barr bodies ≤5% were recorded as male and those with >5% were recorded as female. The percentage accuracy in determining sex using both stains was evaluated. RESULTS: The percentage of Barr bodies in AO-stained slides ranged from 5 to 32 among females and from 0 to 8 in males, while with PAP the ranges recorded were 4-20 in females and 0-5 in males (p < 0.001). The sensitivity and specificity of AO for detecting sex accurately was around 98.3 and 95% for PAP. CONCLUSION: Sex determination using Barr bodies in the buccal cells is a simple method that provides up to 95-98% accuracy; making it a significant accessory for sex determination. AO proved to be a better stain than PAP for visualizing nuclear details, and its staining time was remarkably shorter. It also demonstrated enhanced sex estimation efficacy compared to PAP.

Expression of transforming growth factor ß and its correlation with lipodystrophy in oral submucous fibrosis: an immunohistochemical study.

OBJECTIVE: In our routine review of Oral Submucous Fibrosis (OSMF) biopsies, we observed decreased adipose tissue even though most are from buccal mucosa. Pathogenesis of OSMF has demonstrated the role of Transforming Growth Factor ß (TGF ß), in causing fibrosis. This study aims to correlate the role of TGF ß with loss of adipose tissue in OSMF. STUDY DESIGN: From our archives, 84 OSMF cases (24 early and 60 advanced OSMF) were screened for adipose tissue. Immunoexpression of TGF ß in these cases were investigated. RESULTS: Adipose tissue was seen in 67% of early OSMF and in 13% of advanced cases. Early cases showed more intense TGF ß staining of epithelium, fibroblast, macrophages and inflammatory cells than the advanced cases. CONCLUSION: These findings suggest that TGF ß plays a key role in causing lipodystrophy in OSMF and is secreted more during early course of the disease than in advanced stage.

Evaluation of Presence of Enterococcus faecalis in Root Canals of Deciduous Molars with Necrotic Pulp by Agar Culture and Polymerase Chain Reaction.

Objective: To assess the presence of Enterococcus faecalis in root canals of deciduous molars with necrotic pulp by agar culture and polymerase chain reaction (PCR) assay. Materials and methods: This is an experimental study, where a total of 120 endodontic samples were taken from deciduous molars with necrotic pulps. The presence of Enterococcus faecalis was assessed by culture, using Enterococcus confirmatory agar, and by PCR assay. Statistical analysis of the results was performed using McNemar's test. Results: The presence of Enterococcus faecalis was detected in 20 samples (16.67% of total) by microbial culture and in 45 samples (37.5% of total) by PCR assay, with a statistically significant difference between the two methods (p < 0.001). Microbial culture and PCR both detect Enterococcus faecalis, with the latter detecting an additional 25 positive samples. Conclusion: In this study, PCR assay was significantly more sensitive than agar culture method in detecting the presence of Enterococcus faecalis in root canals of deciduous molars with necrotic pulp, that is, 37.5% of all samples. Clinical significance: Importance of presence of Enterococcus faecalis in necrotic pulps of deciduous teeth, as it is primarily responsible for failure of endodontic treatment, thus helping clinicians to advocate the use of local drug delivery in primary teeth endodontics and also aids clinicians in choosing the most effective intracanal medication. How to cite this article: Nalawade TM, Bhat KG, Kale AD, et al. Evaluation of Presence of Enterococcus faecalis in Root Canals of Deciduous Molars with Necrotic Pulp by Agar Culture and Polymerase Chain Reaction. Int J Clin Pediatr Dent 2023;16(6):816-819.

Sex assessment using odontometry and cranial anthropometry: evaluation in an Indian sample.

Crania and teeth are considered to be useful adjuncts for sex assessment and in construction of a postmortem profile, however, there is very little information regarding the relationship between tooth and cranial size. The purpose of this study was to demonstrate the extent of sexual dimorphism of teeth and cranial size in an adult Indian population and their potential in sex estimation using logistic regression analysis. The sample consisted of 200 subjects (96 males and 104 females; age range of 18-30 years) of Indian origin. Cranial anthropometric measurements i.e. maximum head length and head breadth were measured and cephalic index was calculated. Tooth size (maximum mesiodistal and buccolingual dimensions) was measured for all the permanent teeth of the right side of the maxillary and mandibular arches, except the third molars. To ascertain the usefulness of absolute measurements of crania and teeth and the combination of both these parameters in sex prediction, logistic regression analysis was done. The cranial anthropometric measurements gave a sex assessment accuracy ranging from 53.5 to 79.9%, with head length giving the best accuracy. The sex classification accuracy of the odontometric measurements ranged from 61.5 to 76%, with combination of maxillary and mandibular teeth giving better accuracy. The use of both these parameters together vastly improved the accuracy to 88.4%. This study demonstrates that cranial anthropometry along with odontometry could be used for determining the gender of adult Indians in a forensic context. Among all the parameters, head length gave a superior sex prediction alone (79.9%) as well as in combination with odontometry (88.4%).

Evaluation of myofibroblasts in oral submucous fibrosis: correlation with disease severity.

BACKGROUND: Oral submucous fibrosis (OSMF) is a chronic debilitating disease and a premalignant condition of the oral cavity characterized by generalized submucosal fibrosis. Myofibroblasts are contractile cells expressing α-smooth muscle actin (α-SMA) and are considered primary producers of extracellular matrix after injury. Their accumulation has been established as a marker of progressive fibrosis in organs like lungs, liver, kidney and skin. This study aims to evaluate the presence of myofibroblasts in various histological stages of OSMF. MATERIALS AND METHOD: Seventy cases of OSMF, which were further categorized histologically into early (35 cases) and advanced (35 cases), were subjected to immunohistochemistry using α-SMA antibody for detection of myofibroblasts. Fifteen normal oral mucosa specimens were also stained as controls. RESULTS: The number of α-SMA-stained myofibroblasts in OSMF was significantly increased when compared to that of the normal controls (P<0.001). Additionally, a statistically significant increase in the myofibroblasts population between early and advanced stages was observed (P=0.000). CONCLUSIONS: Our results corroborate the possibility that OSMF actually represents an abnormal healing process in response to chronic mechanical and chemical irritation because of areca nut chewing as demonstrated by the increased incidence of myofibroblasts in this disease. Furthermore, the progressive increase in myofibroblasts from early to advanced stages suggests their potential use as markers for evaluating the severity of OSMF.

Isolation, culture and characterization of primary cell lines of human buccal mucosal fibroblasts: A combination of explant enzamytic technique.

BACKGROUND: The cell culture technique has become a routine and a popular method for its wide applications in the field of cell biology and biotechnology and in medical research. Isolation of primary cells over the cancer cells is an essential component of cell culture technology as they are the reliable source to understand normal physiological, morphological and molecular process of human cells. As fibroblasts are the prominent cells of the connective tissue of oral mucosa, many disease entities and histogenesis are linked to fibroblasts. Culture of oral fibroblast cells helps the oral biologists and researchers to study the morphological and molecular process in the oral diseases. AIM: The aim of our experiment is to isolate and culture the human buccal mucosal fibroblast cells from healthy individuals using a combination of explant-enzymatic method and characterization of the cells by short tandem repeat (STR) profiling. MATERIALS AND METHODS: The tissue samples were collected from healthy individuals undergoing routine impacted third molar extraction. A combination of explant-enzymatic technique was used for the isolation from the tissue samples. The cells were further subcultured, maintained and stored as per the standard protocols. Thus, to confirm the oral fibroblasts of human origin and its uniqueness, they were characterized using STR profiling. RESULTS AND CONCLUSION: Using the combination technique, we were successful in isolating the cells at a faster rate by detachment of cells on day 3 and confluency on day 10. The morphological assessment and STR profiling further confirmed that the isolated cell lines resemble human fibroblast cells.

Roles of the matricellular protein Tenascin-C in T-lymphocyte trafficking and etiopathogenesis of Oral Lichen Planus.

OBJECTIVE: This study was aimed at examining the role of Tenascin-C in T cell trafficking in Oral Lichen Planus (OLP). DESIGN: For the in vivo immunohistochemical analyses, 115 OLP samples were collected from patients and immunostaining was performed. The intensity and distribution of TN-C expression were quantified and correlated with histological analyses of basement membrane integrity and presence of inflammatory infiltrate. For the in vitro study, TN-C and collagen were coated on culture plates and migration of T lymphocytes was assessed. RESULTS: TN-C immunoexpression was increased in terms of both distribution and intensity along the basement membrane zone. These changes were significantly associated with basement membrane duplication (distribution p < 0.002 and intensity p < 0.001) and bands of inflammation (distribution p < 0.002 and intensity p < 0.001) assessed by Chi-square test. T lymphocytes demonstrated significant migration towards TN-C as compared to collagen (n = 3, p < 0.05). CONCLUSIONS: These findings indicate TN-C may have a key role in promoting T cell migration at the epithelial-mesenchymal junction in OLP. These observations suggest TN-C could be a good target for therapeutic intervention, either in itself or synergistically with anti-inflammatory directed strategies in this chronic disease management.

Tumor budding is a potential histopathological marker in the prognosis of oral squamous cell carcinoma: Current status and future prospects.

Tumor budding (TB) is a histopathologically evident feature that represents a scattered pattern of invasion consisting of isolated single tumor epithelial cells or tumor cells in small clusters (up to 5 cells) seen primarily at the invasive front dispersed within the stroma for variable distance. Presence of TB has been linked with lymph node metastasis, recurrence, distant metastasis and reduced survival in numerous cancers including oral squamous cell carcinoma (OSCC). Tumor buds are also considered as histopathological markers of epithelial-mesenchymal transition which is a molecular process implicated as a hallmark for invasion and metastasis. This review gives an overview of the current evidence regarding TB assessment in OSCC and its future prospects.

Stature estimation from regression analysis of facial anthropometry in Indian population.

BACKGROUND: Stature or body height is one of the most important and useful anthropometric parameters which determines the physical identity of an individual. As craniofacial structures have the advantage of being composed largely of hard tissue which is relatively indestructible, the careful study of these can enable reliable determination of stature of the person in life. Studies pertaining to stature estimation from facial measurements are limited in an Indian population. The present investigation attempts to estimate stature from anthropometric dimensions of face. MATERIALS AND METHODS: The material for the present study comprises 361 Indian students (151 males and 210 females) in the age range of 21-45 years. Stature and six facial measurements were taken on each participant following standard methods and techniques. Karl Pearson's correlation coefficient and linear regression were done to estimate stature. RESULTS: The results indicate that facial measurements are strongly and positively correlated (P < 0.001) with stature. The accuracy of the computed equations was further tested on 50 randomly selected study participants of each group, which shows close approximation of actual and estimated stature. CONCLUSION: Within the limits of this study, we conclude that facial dimensions can be used as a supplementary approach for the estimation of stature but with caution, as these are population-specific approach.

Assessment of morphological parameters associated with neural invasion in oral squamous cell carcinoma.

AIMS: Neural invasion (NI) is a form of tumor spread distinct from lymphatic and vascular invasion. It has been correlated with aggressive behavior, disease recurrence and increased morbidity and mortality. Despite the importance of NI as a prognostic indicator, the mechanism and associated factors are poorly understood. Hence, the aim of this study was to determine morphological parameters associated with NI in oral squamous cell carcinoma (OSCC). METHODOLOGY: One hundred and five archival specimens of patients with primary OSCC who underwent surgical excision and radical neck dissection were included in the study. The presence of NI was analyzed in slides and correlated with clinical as well as morphological parameters using univariate and multivariate analysis. RESULTS: NI was identified in 31 cases (29%) of OSCC. NI was significantly associated with tobacco habit, lymphovascular invasion and positive surgical margins. Multivariate analysis further emphasized these factors to be significant risk factors for NI. Peri-NI and intra-NI significantly associated with the size of the tumor, while the distance of invasion was associated with advanced stage. CONCLUSION: NI is associated with multiple morphological parameters and its identification may have a significant impact on the management and prognostic evaluation of OSCC.

Assessment of cell block technique in head and neck pathology diagnoses: A preliminary study.

OBJECTIVE: The head and neck region is a composite site made of multiple tissue components. These tissues when affected by disease or pathology present with an array of changes in the tissue architecture and pattern. It is essential to visualize the cellular details and tissue patterns for accurate diagnosis and treatment planning. Aspiration cytology primarily makes use of the cellular details for diagnosing lesions of the head and neck. Despite the promising results, its use is still limited in certain cases of the head and neck. The reason implicated could be the indiscernible appearance of cells in the absence of tissue integrity. In this regard, cell blocks are known to facilitate the visualization of the cytomorphological as well as the tissue arrangement patterns. Thus, the present study was designed to evaluate the role of cell block cytology in the diagnosis of various lesions of the head and neck. METHODS: Odontogenic lesions, epithelial carcinomas and connective tissue pathology of the head and neck origin were included in the study (n = 45). Aspiration cytology smears and cell block diagnosis were compared with tissue biopsy diagnosis for determining their sensitivity (%) and diagnostic efficacy. RESULTS: Cell blocks showed distinct preservation of the architectural pattern. In case of fluid-filled lesions, the contents were preserved and correlated with the tissue biopsy results. The results of cell blocks were similar to that of tissue biopsy in majority of the cases (95.56%). CONCLUSION: We recommend using cell blocks as a part of routine laboratory practice for all head-neck cases.

Role of immunoreactive patterns of lymph nodes in neck dissection cases of oral squamous cell carcinoma: a clinical and histopathological study.

OBJECTIVES: Metastasis in oral squamous cell carcinoma (OSCC) can occur in a variety of ways, and draining lymphatics and lymph nodes serve as a common route. Prior to metastasis, lymph nodes elicit an immune response to either wall off or create a favorable environment for homing of tumor cells. This immune response to tumor stimuli is visualized by recognizing various immunoreactive patterns exhibited by the lymph node. The present study aims to evaluate the role of immuno-morphologic patterns of the lymph node in neck dissection for cases of OSCC. MATERIALS AND METHODS: Our retrospective study included 50 neck dissection cases of OSCC and a total of 1,078 lymph nodes. The grades of primary tumors with eight different immunoreactive patterns were compared. Vascularity and metastasis in lymph nodes were also evaluated. RESULTS: The lymphocyte predominant pattern was the most common immunoreactive pattern found in 396 of 1,078 lymph nodes. Patterns of lymphocyte predominant (P=0.0005), sinus histiocytosis (P=0.0500), paracortical hyperplasia (P=0.0001), cortical hyperplasia (P=0.0001), and increased vascularity (P=0.0190) were significantly associated with tumor grade. CONCLUSION: The present study adds to the understanding of lymph node immunoreactivity patterns and their correlation with tumor grade. We recommend further study of lymph node patterns for all sentinel lymph node biopsies and routine neck dissections for OSCCs.

Modified Alcohol-Formalin Cell Block Technique in Head and Neck Pathology Diagnosis.

OBJECTIVES: Cell block preparation is a recognized technique in histopathological diagnosis. This technique aids in maintaining an intact architecture and reducing the diagnostic errors associated with fine-needle aspiration cytology. Numerous techniques have been put forth, but the need for an optimal technique for routine use in the laboratory still persists. The aim of our study was to establish a cell block technique which aids in the accurate diagnosis of head and neck pathology. METHODOLOGY: A modified cell block technique was developed using alcohol-formalin as a fixative. Forty fine-needle aspiration fluids from clinically and radiologically diagnosed cases of head and neck pathology were used as samples. The cell block sections were compared with the cytology smears to determine the utility of the technique. RESULTS: The cell blocks presented with better preservation of the architectural framework and enabled a quick diagnosis. Cellular clumping was negligible, and nuclear as well as cellular details were maintained similar to tissue sections. It led to the integration of conventional techniques using 10% neutral buffered formalin with that of the 10% alcohol-formalin technique. CONCLUSION: Modified cell block technique can be used as a simple and effective tool in the routine diagnosis of head and neck pathology.

Comparison of staining of mitotic figures by haematoxylin and eosin-and crystal violet stains, in oral epithelial dysplasia and squamous cell carcinoma.

UNLABELLED: Mitosis of cells gives rise to tissue integrity. Defects during mitosis bring about abnormalities. Excessive proliferation of cells due to increased mitosis is one such outcome, which is the hallmark in precancer and cancer. The localization of proliferating cells or their precursors may not be obvious and easy. Establishing an easy way to distinguish these mitotic cells will help in grading and understanding their biological potential. Although immunohistochemistry is an advanced method in use, the cost and time factor makes it less feasible for many laboratories. Selective histochemical stains like toluidine blue, giemsa and crystal violet have been used in tissues including the developing brain, neural tissue and skin. AIM OF THE STUDY: 1) To compare the staining of mitotic cells in haematoxylin and eosin with that in crystal violet. 2) To compare the number of mitotic figures present in normal oral mucosa, epithelial dysplasia and oral squamous cell carcinoma in crystal violet-stained sections with that in H and E-stained sections. MATERIALS AND METHODS: Ten tissues of normal oral mucosa and 15 tissues each of oral epithelial dysplasia seen in tobacco-associated leukoplakia and squamous cell carcinoma were studied to evaluate the selectivity of 1% crystal violet for mitotic figures. The staining was compared with standard H and E staining. Statistical analysis was done using Mann-Whitney U test. RESULTS: A statistically significant increase in the mean mitotic count was observed in crystal violet-stained sections of epithelial dysplasia as compared to the H and E-stained sections (p=0.0327). A similar increase in the mitotic counts was noted in crystal violet-stained sections of oral squamous cell carcinoma as compared to the H and E-stained sections.(p=0.0443). No significant difference was found in the mitotic counts determined in dysplasia or carcinoma by either the crystal violet (p=0.4429) or the H and E-staining techniques (p=0.2717). CONCLUSION: One per cent crystal violet provides a definite advantage over the H and E-stained sections in selectively staining the mitotic figures.

Immunoexpression of programmed cell death 4 protein in normal oral mucosa, oral epithelial dysplasia and oral squamous cell carcinoma.

BACKGROUND: Oral squamous cell carcinoma (OSCC) is the frequently reported cancer of the head and neck. Recent studies are being conducted to evaluate the role of potential markers for diagnosing the stages of development of OSCC from normal cells. AIM: The aim of this study is to evaluate and compare the immunoexpression of programmed cell death 4 (PDCD4) protein in normal oral mucosa, oral epithelial dysplasia (OED) and OSCC. MATERIALS AND METHODS: Histologically diagnosed, formalin-fixed paraffin-embedded archived cases (n = 100) of normal mucosa (n = 10), OED (n = 60) and OSCC (n = 30) were analyzed immunohistochemically in the present retrospective study using monoclonal rabbit antihuman PDCD4. OED and squamous cell carcinoma were graded according to the World Health Organization and Broder's histological grading criteria, respectively. Clinical parameters and immunohistochemical results were analyzed by Fisher exact test using SPSS software. P <0.05 was indicative of significant differences. RESULTS: PDCD4 expression was observed in the normal oral mucosa, OED and OSCC. The maximum expression was observed in the normal oral mucosa, which reduced significantly in OED and OSCC (P = 0.017). With the increase in the transformation from normal cells to cancer cells, a shift from nuclear to cytoplasmic staining was observed indicating predominant cytoplasmic localization of stain as a feature of altered cells. CONCLUSION: The present study delineates the molecular difference between the normal, dysplastic and carcinomatous cells; and points toward the role of PDCD4 localization in the proliferation of cells. This study thus highlights the need for further research with inclusion of long follow-up period and other pathological criteria such as inflammation and microenvironment, immune status of patient and tumor stage, which could aid in the development of prospective diagnostic options.

Validation of immunoexpression of tenascin-C in oral precancerous and cancerous tissues using ImageJ analysis with novel immunohistochemistry profiler plugin: An immunohistochemical quantitative analysis.

BACKGROUND: Immunohistochemistry (IHC) is a molecular technique that has grown tremendously over the years. However, the assessment is only qualitative which is subjective and causes errors. Due to this limitation, several excellent markers have not gained importance and reached clinical trials. Hence, we aimed to quantify IHC by ImageJ analysis with a novel IHC profiler plugin. ImageJ has not been tried in oral precancerous tissues with minimal attempt for matrix markers. AIM: This study aimed to validate the quantification of immunoexpression of tenascin-C (TN-C) in oral precancerous tissues and oral squamous cell carcinoma (OSCC) using ImageJ software with IHC profiler plugin. MATERIALS AND METHODS: After IHC staining for TN-C and image acquisition, ImageJ analysis was performed as per the standard recommended algorithm. Assessment was done by two observers by blinding the histopathological diagnosis. The immunoscore was assessed for interobserver variability using Kohen's kappa statistics. RESULTS: All our cases were in agreement and found to be statistically significant with P < 0.005. Moderate agreement was for mild dysplasia, moderate dysplasia and oral lichen planus. Substantial agreement was for oral submucous fibrosis and OSCC and almost perfect agreement noted for cases of severe dysplasia. CONCLUSION: IHC can now be quantified using freely downloadable software ImageJ analysis in oral precancerous tissues and OSCC. This software with good threshold control can quantify matrix marker such as TN-C. Hence, herewith, we propose that IHC markers should be quantified using ImageJ by our entire oral pathology fraternity so as to have a standard immunoscore for all markers.