Formulation and Development of a Herbal Antifungal Gel Containing Origanum vulgare and Syzygium aromaticum Essential Oils Against Oral Candida albicans.

Background Oral candidiasis is the most prevalent oral fungal infection, and existing antifungal agents have side effects such as drug intolerance, resistance, and toxicity. Herbal essential oils are emerging as an alternative therapeutic approach for treating fungal infections. Origanum vulgare (O. vulgare), commonly known as oregano, and Syzygium aromaticum (S. aromaticum), commonly known as clove, are known to have antifungal properties and are effective against fluconazole-resistant strains. A combination of essential oils has a synergistic effect and aids in achieving effective antifungal activity at sufficiently low concentrations, which could lead to reduced side effects and resistance. Aim of the study This study aimed to formulate and develop an herbal antifungal gel containing O. vulgare and S. aromaticum and evaluate its synergistic antifungal efficacy against oral Candida albicans (C. albicans). Methodology Minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) determinations of O. vulgare and S. aromaticum essential oils were performed individually and in combination to assess the antifungal activity against C. albicans. Based on the obtained MIC and MFC of essential oils in combination, an herbal antifungal gel was formulated. Further, to determine the biocompatible nature of the gel, a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed. Results We found that a combination of O. vulgare and S. aromaticum essential oils showed antifungal activity at a lesser concentration, with a MIC of 0.19 µl/ml and MFC of 0.39 µl/ml when compared to their individual concentrations. Based on our results, an antifungal herbal gel comprising a concentration of 0.6 µl/ml of both essential oils was developed to achieve synergistic antifungal activity against oral C. albicans. The MTT assay of the herbal gel did not show any cytotoxicity. Conclusion The novel herbal antifungal gel containing O. vulgare and S. aromaticum is biocompatible in nature and provides an alternative therapeutic approach for treating oral candidiasis.

Evaluation of Immunoexpression of AJUBA Protein in Normal Oral Mucosa and Oral Squamous Cell Carcinoma.

AJUBA is multifunctional scaffold protein which belongs to Zyxin family of proteins. It is known to have dual role in cancer as a tumor promoter and tumor suppressor. AJUBA has a key role in systemic malignancies like esophageal squamous cell carcinoma, colorectal cancer, cervical, breast, prostate cancer, etc. But there is very sparse literature available regarding its expression profile in oral squamous cell carcinoma (OSCC) and moreover its expression has not been observed in normal oral mucosa (NOM). Thus, the aim of this research is to explore the expression profile of AJUBA by immunohistochemical method in NOM and OSCC. Furthermore, we also evaluated the association of AJUBA expression with clinicopathologic parameters. A total of 84 samples of formalin fixed paraffin embedded tissue blocks comprising of 42 cases each of NOM and OSCC were subjected to detect immunoexpression of AJUBA. We found enhanced intense immune-expression of AJUBA in OSCC cases than compared with NOM and found to be statistically significant. The parameters specific to histologic tumor grade and inflammatory response in OSCC also found to have statistically significant with AJUBA expression. Our study is first of its kind to reveal AJUBA expression in basal and suprabasal layer of NOM suggestive of its definitive role in differentiation and stratification process. We also observed its intense expression in peripheral cell of tumor islands of OSCC cases, which can suggest its possible role in tumor growth and progression.

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.

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.

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.

Correlation of matrix metalloproteinase-9 expression with morphometric analysis of mucosal vasculature in oral squamous cell carcinoma, oral epithelial dysplasia, and normal oral mucosa.

OBJECTIVE: Oral squamous cell carcinoma (OSCC) invades and metastasizes, by degrading the extracellular matrix (ECM) and is associated with poor prognosis. Matrix-metalloproteinase (MMP-9) can initiate ECM degradation and angiogenesis which brings a significant change in tumor microenvironment favoring tumor progression. A major thrust has been laid on understanding this key enzyme as it has significant implications for cancer therapy. Comprehending the association of vasculature with MMP-9 expression in precancerous lesions oral epithelial dysplasia (OED) and OSCC is essential since the data regarding the same are fewer. The aim is to evaluate and correlate MMP-9 expression with morphometric analysis of mucosal vasculature in the normal oral mucosa (NOM), OED, and OSCC. METHODS: A total of 60 histologically diagnosed cases of OED (n = 30); OSCC (n = 30); along with 10 NOM (n = 10) as control were included. Immunohistochemical staining of MMP-9 and vascular morphometric analysis was performed for all the cases. Results were analyzed using the Chi-square test, Fischer exact test, and Spearman correlation test. RESULTS: A statistically significant difference in MMP-9 was noted among the groups with P = 0.011* (epithelium); P = 0.001* (stroma) by the highest value in OSCC group. Morphometry also revealed a progressive increase from NOM to OED to OSCC. Spearman's correlation of MMP-9 with vascular parameters illustrated a positive relation of MMP-9 with mean vascular density (MVD) and mean vascular area percentage (MVAP). CONCLUSION: Positive correlation of MMP-9 with MVD and MVAP demonstrates this markers effect on angiogenesis. Henceforth, MMP-9 can be embattled as a potential therapeutic target in combating tumor progression.

Immunohistochemical expression of extracellular matrix metalloproteinase inducer (EMMPRIN) in normal oral mucosa, oral epithelial dysplasia and oral squamous cell carcinoma.

BACKGROUND: In oral squamous cell carcinoma (OSCC), extracellular matrix metalloproteinase inducer (EMMPRIN) expression has been noted in the cell membrane throughout the epithelium of the lesion, suggesting its increased expression. OBJECTIVES: The present study was conducted to evaluate and compare the expression of EMMPRIN in the normal oral mucosa (NOM), different histological grades of oral epithelial dysplasia (OED) and OSCC. MATERIALS AND METHODS: A total of 100 formalin-fixed paraffin-embedded tissue blocks of NOM (n = 10), 20 cases each of mild, moderate and severe (OED) (n = 60), and 10 cases each of well differentiated, moderately differentiated and poorly differentiated carcinomas (n = 30) were included in the study. The tissues sections were immunohistochemically stained and were evaluated for intensity and area of expression in different groups. RESULTS: Out of 60 cases of OED, 29 (48%) cases showed intense dark brown staining in the epithelium. The stroma in 38 (63%) cases showed positive immunoexpression. The expression of EMMPRIN in OSCC revealed intense dark brown staining in 9 (90%) cases of well differentiated, and a decent thereon in 8 (80%) cases of moderately differentiated and 4 (40%) cases of poorly differentiated carcinomas. CONCLUSION: The role of EMMPRIN in precarcinogenesis and early carcinogenesis needs to be studied on considerable sample size. This can enable oncologists to detect cancer at an early stage before it progresses to malignancy.

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.

Multiple Mott Cells in Periapical Lesion of the Oral Cavity.

An entity of bizarre plasma cells has been observed, containing spherical inclusions of immunoglobulins within the cytoplasm. These cells are termed as Mott cells and are believed to occur commonly in cases of chronic inflammation. Until now, they were reported to occur in systemic diseases, various hematolymphoid malignancies and in some syndromes; but their occurrence in chronic periapical infections of the oral cavity, given the paucity of available literature, raises questions about their obscure nature. In the present case report, a 24-year male presented with a draining sinus and periapical lesion involving the maxillary right incisors. After clinical and radiographic examination, an excisional biopsy sample of the periapical lesion was obtained. Histopathological examination revealed multiple bizarre Mott cells in the connective tissue stroma. The significance, nature or fate of these Mott cells is still unknown. Frequently overlooked or a chance occurrence, the presence of these multiple Mott cells in chronic inflammatory lesions of the oral cavity could be of some significance and demand increased awareness and further research.

Oral Carcinoma Cuniculatum: A New Entity in the Clinicopathological Spectrum of Oral Squamous Cell Carcinoma.

Carcinoma cuniculatum is principally recognized as a variant of carcinoma involving foot. The World Health Organization (WHO) recognizes Oral Carcinoma Cuniculatum (OCC) as a distinct and rare clinicopathological variant of Oral Squamous Cell Carcinoma (OSCC). OCC is confused clinically and histologically with Verrucous Carcinoma (VC) and is often misdiagnosed as either VC or OSCC. To best of our knowledge, till date, only 50 cases of this tumour have been reported in oral cavity (including the present case) and only limited number of cases have been reported from Indian subcontinent. Pathognomonic feature of OCC is proliferation of stratified squamous epithelium and its infiltration into underlying stroma forming a complex pattern of keratin cores and keratin filled crypts. These complex crypts give it a likeness of rabbit burrow hence, the name cuniculatum (cuniculatus='rabbit warren'). The report aims to present a case of OCC of mandibular gingiva, discuss its diagnostic features and highlight its differences from VC and OSCC.

Immunoexpression of Epithelial Mesenchymal Transition Proteins E-Cadherin, ß-Catenin, and N-Cadherin in Oral Squamous Cell Carcinoma.

Aims Epithelial mesenchymal transition (EMT) is a crucial process for acquisition of malignant phenotype, aggressiveness, and metastatic capacity in neoplasms. It is characterized by loss of epithelial markers and gain of mesenchymal markers. Studies on EMT and its potential association with the histological grading are sparse in oral squamous cell carcinoma (OSCC). This study aims to evaluate the expression of EMT-associated proteins-E-cadherin, ß-catenin, and N-cadherin-in different grades of OSCC. Methodology In all, 60 cases of OSCC further subdivided into 20 cases each of well-, moderately, and poorly differentiated OSCCs were stained immunohistochemically with E-cadherin, ß-catenin, and N-cadherin antibodies. The differences in the expression were evaluated using χ2 and Fisher exact tests, whereas Spearman's correlation was used to analyze the correlation between the markers. Results A reduced E-cadherin expression noted in 40% of the OSCCs was associated with reduced ß-catenin expression in 66.6% of the cases and increase in the expression of mesenchymal N-cadherin seen in 80% of cases. This expression pattern demonstrated a significant association with histological grades. A membrane to cytoplasmic shift of E-cadherin (73.3%) and ß-catenin (78.3%) increased with histological grade. A negative correlation was observed with the E-cadherin and N-cadherin localization, though it did not reach statistical significance. Conclusion OSCC tissues had high levels of EMT phenotype as compared with the normal oral mucosa. This phenotype was characterized by reduced E-cadherin and ß-catenin expression and overexpression of N-cadherin. Aberrant localization of the studied proteins was a hallmark for depicting EMT.

Clinicopathologic study of a series of giant cell fibroma using picrosirius red polarizing microscopy technique.

BACKGROUND: Giant cell fibroma (GCF) is a distinct type of fibroma with characteristic large, stellate mononuclear or multinucleated giant fibroblasts; the stroma of GCF is relatively unexplored. The Picrosirus red polarizing microscopy technique is used to characterize the collagen fibers. The aim of this study was to evaluate the staining properties of collagen fibers in GCF and to correlate it with fibroma using Picrosirius red under the polarizing microscope and van Gieson under bright field microscope. METHODS: In the present study van Gieson and picrosirius red stained slides of 7 cases each of GCF and fibroma were compared for the staining properties of collagen. Using picrosirus red polarizing microscopy; colors noted in fibroma included yellow, yellowish-orange and green, whereas the GCF showed predominantly yellow and orange colors. In Van Gieson stained sections it was observed that the collagen in GCF was densely packed and arranged perpendicular to the epithelium while the collagen in fibroma was loosely packed and arranged parallel to the epithelium. CONCLUSION:   Observable differences in the stroma of the collagen of GCF and fibroma were noted. Collagen in GCF was more mature and dense. The Picrosirius red polarizing technique can be used to assess the collagen in GCF.

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.

Efficacy of stereomicroscope as an aid to histopathological diagnosis.

UNLABELLED: Grossing and microscopic examination of the received specimens in a histopathological laboratory is a routine procedure for achieving a fi nal diagnosis. Errors in either of the steps may lead to an inaccurate diagnosis since wrong orientation of the specimen may either cause diagnostic delays or pose a diagnostic dilemma. Stereomicroscope is an important accessory instrument which can be used to study a variety of specimens. It not only enables us to study the surface details but also aids in minute work including dissection and microsurgery to name a few. AIM: To assess the effectiveness of stereomicroscope in routine grossing and its role in arriving at a final diagnosis. MATERIALS AND METHODS: Cases from the archives of the department for which stereomicroscopic evaluation was done were retrieved and reviewed. RESULTS AND CONCLUSION: Stereomicroscopic images aided not only in the proper orientation of the specimen but also in the diagnosis.

Expression of cytokeratin subtypes: MMP-9, p53, and αSMA to differentiate basaloid squamous cell carcinoma from other basaloid tumors of the oral cavity.

Basaloid squamous cell carcinoma (BSCC) is rare variant of oral squamous cell carcinoma (OSCC) with predilection for upper aerodigestive tract. Although it is characterized by distinct histologic features it is often confused with conventional OSCC and other basaloid tumors. The study aims to establish differentiating features of BSCC with oral basaloid tumors using immunohistochemical (IHC) markers. This retrospective study included 34 cases, including BSCC, OSCC, and basaloid tumors. IHC staining was performed with primary antibodies against cytokeratin (CK) 19, 14, 8/18, α-smooth muscle actin (αSMA), p53, and MMP-9. A prominent CK 19, 14, and 8/18 expression was observed in BSCC as compared with basaloid tumors suggesting of basal cell origin with undifferentiated type of tumor cells. Expression of αSMA was intense in tumor cells of myoepithelial differentiation but lacked in BSCC. The intense expression of p53 and MMP-9 was noted in all basaloid malignancies. Considering standard histologic criteria in diagnosing BSCC, when in confusion with other basaloid tumors, IHC markers gain importance. Hence, enhanced expression of CK 19, 8/18, and 14 and coexistence of p53 and MMP-9 expression and negativity for αSMA suggest an accurate diagnosis of BSCC.

Expression of transforming growth factor â 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 â (TGF â), in causing fibrosis. This study aims to correlate the role of TGF â 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 â 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 â staining of epithelium, fibroblast, macrophages and inflammatory cells than the advanced cases. Conclusion: These findings suggest that TGF â plays a key role in causing lipodystrophy in OSMF and is secreted more during early course of the disease than in advanced stage (AU)

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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.

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.