Salivary and Serum Interleukin-6: A Credible Marker for Predicting Oral Leukoplakia and Oral Squamous Cell Carcinoma by Enzyme-Linked Immunosorbent Assay (ELISA).

BACKGROUND: Oral squamous cell carcinoma (OSCC) is the most prevalent subtype of oral cancer. Detecting oral potentially malignant disorders (OPMDs) in their early stages is crucial to prevent their advancement into OSCC. One of the primary factors contributing to OSCC is tobacco use, which can lead to increased production of cytokines. Among these cytokines, interleukin-6 (IL-6), an immune molecule involved in inflammation, may serve as a valuable indicator for assessing the progression of OPMDs and OSCCs. AIMS: The aim of this study is to assess the levels of IL6 in both serum and saliva using the enzyme-linked immunosorbent assay (ELISA) technique and to determine the prognostic value of these measurements in individuals with oral leukoplakia and OSCC. MATERIALS AND METHODS: The research involved 45 participants, who were categorized into three groups: OSCC (15), leukoplakia (15), and a control group consisting of healthy individuals (15). Saliva and serum samples were collected from each individual within all three groups and analyzed using the ELISA method. Subsequently, the gathered data underwent statistical analysis for evaluation. RESULTS: There were elevated levels of IL-6 in both saliva and serum among individuals with OSCC in comparison to those with leukoplakia and the healthy control group, and this difference was statistically significant. The analysis of ROC (Receiver Operating Characteristic) curves demonstrated that salivary IL-6 was a more effective indicator than serum IL-6 for detecting the advancement of OSCC. As the histological grade of differentiation increased in both OSCC and leukoplakia cases, there was a corresponding rise in salivary IL-6 levels. CONCLUSION: Both salivary and serum IL-6 levels have the potential to serve as valuable prognostic biomarkers for oral leukoplakia and OSCC which shows possible involvement of IL-6 in the development and progression of these conditions. Salivary IL-6 is a superior prognostic marker compared to serum IL-6 due to its non-invasive nature which makes it a useful tool for mass screening.

Eosinophils: An imperative histopathological prognostic indicator for oral squamous cell carcinoma.

BACKGROUND: Inflammation in tumor microenvironment assists in both promotion and growth of tumor. Tumor-associated tissue eosinophilia (TATE) is the term used when eosinophils are observed in a tumor tissue with inflammatory infiltrate. Although carcinogenesis with inflammation is one of the important hallmarks, the exact role of eosinophils remains unclear. Various studies on oral squamous cell carcinoma (OSCC) that focused on eosinophils reported both favorable and unfavorable prognosis in cancer tissue, because of which the exact function of eosinophils still remains uncertain. AIMS AND OBJECTIVES: The present study aims at identifying the role of TATE in OSCC and in malignant transformation of oral epithelial dysplasia (OED). MATERIALS AND METHODS: The study includes 70 samples that divided into two groups, of which 50 histopathologically proven cases of different grades of OSCC and 20 cases of OED (oral leukoplakia). Congo red stain was used to stain the tissue sections. Each slide was viewed under high power in 10 consecutive microscopic fields for counting of eosinophils. RESULTS: Statistical analysis of values obtained was done using ANOVA, unpaired t-test and Mann-Whitney test. The results were statistically significant (P < 0.05) with a mean total eosinophil count of 2.12 in OED and 4.31 in OSCC. CONCLUSION: The present study showed higher eosinophil counts in OSCC when compared to dysplasia which should prompt for a thorough evaluation of tumor front for invasiveness. Therefore, tissue eosinophil count may be used as an adjunct to predict the malignant transformation of dysplastic lesions to OSCC.

Salivary Tumour Necrosis Factor-α as a Biomarker in Oral Leukoplakia and Oral Squamous Cell Carcinoma.

Background: Oral cancer is one of the life threatening disease which requires an availability of a biomarker for its early detection and also for effective treatment strategies. The current study is done to evaluate the efficacy of one such biomarker i.e. TNF- α as an indicator for oral precancer and oral cancer. Objectives: To evaluate the efficacy of Tumour necrosis factor - alpha (TNF)-α as a salivary biomarker in histopathologically diagnosed cases of oral leukoplakia and Oral squamous cell carcinoma. To correlate the levels of TNF- α with varying histologic grading in Oral Squamous Cell Carcinoma and dysplasia grading in Oral leukoplakia or Hyperkeratosis. Materials and Methods: The study group included 90 subjects that were divided into three groups. OSCC (n=30), leukoplakia (n=30) and controls (n=30). Cases were selected based on inclusion and exclusion criteria of the study. Salivary samples were then collected from all three groups. Salivary levels of TNF-α were estimated using Enzyme Linked Immunosorbent Assay (ELISA). The data on concentration gradients obtained were subjected to appropriate statistical analysis. Results: The results of the present study demonstrated higher levels of salivary TNF-α in individuals with OSCC compared to leukoplakia and healthy control subjects with a high level of statistical significance. ROC curve analysis along with diagnostic parameter calculation also revealed that salivary TNF-α to be a better medium for detecting OSCC. There is also an increase in the salivary TNF-α levels with increase in the histological grade of differentiation in OSCC as well as leukoplakia. Conclusion: The present study concludes that salivary TNF ­ α can be used as a prognostic biomarker of OSCC. In view of the elevated levels of TNF ­ α in saliva of individuals with severe dysplasia, it can also be used to monitor the malignant transformation to leukoplakia to OSCC.

Evaluation of Proliferative Marker Ki-67 in Adenoid Cystic Carcinoma: A Retrospective Study.

AIM: Adenoid cystic carcinoma (ACC) is a malignant tumor of salivary gland origin. Although the histologic appearance of ACC is low grade, management of this malignancy is a distinct therapeutic challenge because of its tendency for perineural involvement and potential for distant metastasis. Ki-67 antigen is expressed during the G1, S, G2 and M phases in the cell cycle but is absent in the quiescent G0 phase in tissue sections. Aim of the study was to review hematoxylin and eosin stained slides in order to confirm the previous histopathological diagnosis as per the criteria given by World Health Organisation (WHO) and to evaluate the expression of cell proliferation marker, Ki-67 antigen in Adenoid cystic carcinoma and correlate the expression of Ki-67 antigen histopathologically with different grades in Adenoid cystic carcinoma. MATERIALS AND METHODS: Tissue samples of 32 cases (12 males and 20 females) were selected from minor salivary glands with age range from 21 to 70 years. Two paraffin-embedded sections of these total 32 cases each of 4 µm thick were cut on a rotary microtome. One section was stained using hematoxylin and Eosin (H&E) and the other was used for Immunohistochemical staining with Ki-67 antigen. RESULTS: Among these 32 cases of Adenoid cystic carcinoma, Histologically 14 (43.75%) tumors were classified in grade I, 8 (25%) were in grade II, and 10 (31.25%) were in grade III. The average percentage of Ki-67 expression was 27.12% in grade I, 34.43% in grade II and 38.45% in grade III. CONCLUSION: Ki-67 immunoreactivity increased with increase in histopathological grades of ACC. CLINICAL SIGNIFICANCE: Since Ki-67 is a useful marker for assessing the proliferative potential of tumors, the prognosis of patients can definitely be predicted.

Calcifying cystic odontogenic tumor associated with ameloblastoma - A rare histological variant.

Calcifying odontogenic cysts (COCs) represent a heterogeneous group of lesions that exhibits a variety of clinical, behavioral, and histological features. COC or Gorlin's cyst is now classified by the World Health Organization as a tumor. Calcifying cystic odontogenic tumor (CCOT) is an uncommon developmental odontogenic lesion that demonstrates histopathologic diversity. Odontogenic tumors such as ameloblastoma have been reported to be associated with CCOT. In this paper, we report a case of ameloblastomatous CCOT in a 16-year-old girl with involvement of mandibular posterior region, which is an extremely rare histologic variant.

Protein 53, B-Cell Lymphoma-2, Cyclooxygenase-2, and CD105 Reactivity in Keratocystic Odontogenic Tumors: An Immunohistochemical Analysis.

OBJECTIVE: The objective is to evaluate and compare immunohistochemically, the biological behavior of keratocystic odontogenic tumor (KCOT) with normal oral mucosa by analyzing cell proliferation, angiogenesis, and antiapoptosis using cyclooxygenase-2 (COX-2), protein 53 (p53), B-cell lymphoma-2 (Bcl-2), and CD105 (endoglin). MATERIALS AND METHODS: The present study comprised 30 cases of KCOT in the study group, in which the expression of COX-2, p53, Bcl-2, and CD105 were analyzed and compared with that of control group consisting of 30 normal oral mucosae. RESULTS: The results were evaluated based on a number of positively stained cells; among 30 samples of KCOT, 22 samples were p53 positive, 23 samples were Bcl-2 positive, 18 samples were COX-2 positive, whereas all the above markers were negative in all the 30 samples of the normal oral mucosa and CD105 mean vascular density in KCOT 13.8 in normal oral mucosa. CONCLUSION: The present study suggests that angiogenesis, cell proliferation, and antiapoptosis may be the possible factors contributing for the unique biological behavior of KCOT.

Molecular Detection of Porphyromonas gingivalis in Chronic Periodontitis Patients.

AIM: In the current study, Porphyromonas gingivalis was identified in chronic periodontitis patients and healthy subjects by polymerase chain reaction (PCR) and its presence correlated with the severity of clinical periodontal parameters. MATERIALS AND METHODS: Subgingival plaque samples were collected with sterile curette and subjected to deoxyribonucleic acid (DNA) extraction and subsequent PCR for detection of P. gingivalis. RESULTS: Porphyromonas gingivalis was detected in 60% of patients of group II (pocket depth up to 5 mm), and in 93.33% of patients of group III (pocket depth more than 5 mm). One periodontally healthy subject in group I (probing depth < 3 mm) showed the presence of P. gingivalis. CONCLUSION: Detection frequency of bacterium increased significantly with increase in probing pocket depth (PPD), loss of attachment (LOA), and gingival index (GI). CLINICAL SIGNIFICANCE: Porphyromonas gingivalis is strongly associated with chronic periodontitis and its detection frequency positively correlates with the severity of periodontal destruction.

Role of cell proliferation and vascularity in malignant transformation of potentially malignant disorders.

BACKGROUND: Significant increase in cell proliferation and vascularity occurs during the transition from normal oral mucosa through differing degrees of dysplasia to oral squamous cell carcinoma (OSCC). AIMS: To evaluate the cell proliferation and vascularity in potentially malignant disorders and OSCC. SETTINGS AND DESIGN: Proliferating cell nuclear antigen (PCNA), vascular endothelial growth factor (VEGF) and CD34 were quantified immunohistochemically (IHC) using anti-PCNA, anti-VEGF and anti-CD34 antibody. MATERIALS AND METHODS: A total of 60 archival specimens included 10 oral lichen planus, 10 oral leukoplakia, 10 oral submucous fibrosis and 30 OSCC (well differentiated, moderately differentiated and poorly differentiated), and also, 10 normal oral mucosa as control group were taken. PCNA, VEGF and CD34 expression was assessed in relation to the localization and area of IHC-stained cells. STATISTICAL ANALYSIS: One-way analysis of variance test and post hoc least significant difference test were assessed for statistical significance. RESULTS: Cell proliferation and vascularity appeared to increase gradually with disease progression. CONCLUSION: Upregulation of cell proliferation and vascularity indicates their possible role in malignant transformation of potentially malignant disorders.

Cytological Changes in Normal Oral Mucosa of Individuals with Tobacco Habits: A Cytomorphometric Study.

INTRODUCTION: Oral cancer is one of the six most common cancers in the world, and globally more than 50% of head and neck cancers occur in Asia, remarkably in India. Overall, 200,000 cases of head and neck cancers occur each year in India, among which 80,000 are oral cancers. Epidemiological and clinical studies suggest a causative role of tobacco use in the evolution of oral potentially malignant and malignant disorders. AIMS AND OBJECTIVES: The aim of the study is to evaluate independently and compare the cytological effects of smoking, tobacco chewing, and smoking in conjunction with tobacco chewing on oral mucosa by cytomorphometric analysis. MATERIALS AND METHODS: The study included a total of 120 individuals subdivided into four groups, each group with 30 individuals. Group I was tobacco smokers, group II tobacco chewers, and group III both tobacco smokers and chewers. Group IV comprised 30 individuals without tobacco habit. Smears were prepared from buccal mucosa of both the study and control groups using a cytobrush and stained using Papanicolaou staining. The cells were quantified using image analysis software. RESULTS: The results of the study showed alterations in the nuclear and cellular parameters in the study groups when compared with control groups and were statistically significant (p < 0.05). CONCLUSION: The present study explains the significance of early identification of cellular changes in individuals with tobacco habits who require early intercession even without any visible oral mucosal changes. The study emphasizes that exfoliative cytology and cytomorphometry aid as a valuable tool to evaluate the effect of tobacco on oral mucosa. CLINICAL SIGNIFICANCE: Simple noninvasive techniques like exfoliative cytology can be employed as a chairside technique and in mass screening programs for identification of cellular changes in oral mucosa of individuals with tobacco habits. Thereby, it can be used as an early diagnostic tool for identification of potentially cancerous and cancerous lesions.

Honey and olive oil as bio-friendly substitutes for formalin and xylene in routine histopathology.

BACKGROUND: Formalin has long been the standard fixative and xylene has been the clearing agent for routine histopathology and immunohistochemistry worldwide. In recent years, as a result of increasing concerns about the potential carcinogenicity of formaldehyde and xylene, attempts have been made to find safer alternatives. In the present study, we considered honey as better alternative for formalin and olive oil as safer substitute for xylene. AIMS: The aim of this study was to know whether honey could be a possible substitute for formalin and olive oil could be a possible substitute for xylene. MATERIALS AND METHODS: Thirty routine biopsy tissues of 1-2 cm were taken. The study group was divided into Group A and Group B. Group A were subjected to normal processing. Group B were fixed into honey for 24 h after which it was taken through routine processing, and then immersed in olive oil instead of xylene. All the sections will be stained with routine hematoxylin and eosin staining. Compare the sections of both the methods. RESULTS AND CONCLUSION: The preservation of tissue by honey giving superior result when compared to that of formalin. Olive oil was found to be effective clearing agent compared to xylene.

Collagen and Its Role in predicting the Biological Behavior of Odontogenic Lesions.

INTRODUCTION: Odontogenic cysts and tumors have variable recurrence rates. Recurrence rate is mainly due to the activity of the epithelium. The epithelium of these lesions has been investigated extensively in regard to their role in proliferative and aggressive behavior of the lesions. However, the role of the connective tissue wall in their behavior has not been studied as extensively. Collagen is an essential part of the connective tissue as a whole and fibrous wall of cystic lesions especially. It is demonstrated by picrosirius red dye staining combined with polarization microscopy. This method permits the evaluation of the nature of the collagen fibers in addition to their thickness. MATERIALS AND METHODS: A total of 56 histopathologically diagnosed cases comprising odontogenic follicle, dentigerous cyst, unicystic ameloblastoma, keratocystic odontogenic tumor (KCOT), multicystic/solid ameloblastoma, and ameloblastic carcinoma were taken and stained using picrosirius red stain and evaluated using a polarizing microscope. RESULTS: Collagen fibers in odontogenic follicles and dentiger-ous cysts showed predominant orange-red birefringence; fibers in unicystic ameloblastoma and KCOT showed both orange red and greenish-yellow birefringence; and fibers of multicystic/ solid ameloblastoma showed predominant greenish-yellow birefringence and ameloblastic carcinoma that showed almost complete greenish birefringence. As the biological behavior of the lesions in the spectrum studied progress toward aggressive nature, increase in immature collagen fibers is noticed. CONCLUSION: This study suggests that the nature of collagen fibers plays a pivotal role in predicting the biological behavior of odontogenic lesions. CLINICAL SIGNIFICANCE: Aggressive nature of the odontogenic lesions is determined by both the epithelium and the connective tissue components (collagen). Studying the nature and type of collagen helps in predicting its biological behavior.

Cementum as an age determinant: A forensic view.

CONTEXT: Forensic age estimation (FAE) defines an expertise in forensic medicine, which aims to define in the most accurate way to determine the unknown chronological age of the person involved in judicial or legal proceedings. Dental cementum is a vital tissue which demonstrates continuous apposition throughout the life of the tooth. This appositional changes of cementum helps in approximation of age inforensic investigations. AIMS: To correlate age by measuring the overlap or coronal migration of thecementum at thecementoenamel junction (CEJ) and the thickness of the cementum at the apical third of the root. SETTINGS AND DESIGN: A hundred freshly extracted teethfrom patients ranging from ages 17-55were longitudinal buccolingually ground sectioned using a mounted lathe wheel and Arkansas stone. MATERIALS AND METHODS: 100 freshly extracted teeth of age group ranging from 17-55 years were taken. These teeth were longitudinally ground sectioned to a thickness of 8-10µm using a mounted lathe wheel and Arkansas stone. Afterwards the teeth were examined under a light microscope using a micrometer eyepiece for measuring the overlap or coronal migration of the cementum at the CEJ and the thickness of the cementum at the apical one-third of root. STATISTICAL ANALYSIS: Measurements of the overlap or the coronal migration of the cementum at the CEJ and the thickness of the cementum at the apical one-third of the root are correlated with age. RESULTS: Results of the study indicated that the cementum at the CEJ migrated coronally during theaging process in case of the impacted teeth. There is also a significant increase in the thickness of the cementum at the apical onethird of rootin the case of both the impacted and erupted teeth. CONCLUSION: Approximation of age by measuring overlap or coronal migration of the cementum at the CEJ and the thickness of the cementum at the apical one-third of the rootsets new alleys in FAE.

Bio-Friendly Alternatives for Xylene - Carrot oil, Olive oil, Pine oil, Rose oil.

BACKGROUND: Xylene is a flammable liquid with characteristic petroleum or aromatic odours, it is miscible with most of the organic solvents and paraffin wax. Xylene clears tissues rapidly and renders transparency, facilitating clearing endpoint determination, this made it to be used as a clearing agent in routine histopathological techniques. Even though it is a good clearing agent, it causes damage to the tissues by its hardening effect particularly those fixed in non-protein coagulant fixatives. Apart from these tissue effects, it has severe, long lasting ill effects on health of technicians and pathologists when exposed to longer duration. Hence in order to overcome these effects and replace xylene with a safe alternative agent, the present study was carried out to assess the clearing ability and bio-friendly nature of four different natural oils i.e., Carrot oil, Olive oil, Pine oil and Rose oil in comparison with that of Xylene. According to Bernoulli's principle of fluid dynamics, to decrease viscosity of these oils and increase penetration into tissues for rapid clearing hot-air oven technique was used. AIMS: To assess:1) Clearing ability and bio-friendly nature of four different oils i.e., Carrot oil, Olive oil, Pine oil, Rose oil in comparison with that of xylene, 2) Application of Bernoulli's principle of fluid dynamics in rapid clearing of tissues by using hot-air oven. MATERIALS AND METHODS: Forty different formalin fixed tissue samples were taken. Each sample of tissue was cut into 5 bits (40x5=200 total bits) which were subjected for dehydration in differential alcohol gradients. Later, each bit is kept in 4 different oils such as Carrot oil, Olive oil, Pine oil, Rose oil and xylene and transferred into hot-air oven. Further routine steps of processing, sectioning and staining were done. Individual sections cleared in four different oils were assessed for cellular architecture, staining quality and a comparison was done between them. RESULTS: Results showed that all the four oils had ability to clear the tissues similar to that of xylene. Pine oil was superior in its physical and clearing properties, which also maintained good cellular architecture and distinct staining quality, when compared with other oils and xylene. CONCLUSION: Carrot oil, Olive oil, Pine oil, Rose oil, are not only bio friendly and economical but can also be used as clearing agent instead of xylene. In this study stability and longetivity of haematoxyllin and eosin staining was evaluated for over a period of one year and no significant difference in staining quality were observed.

Oral Cysticercosis- A Diagnostic Dilemma.

Cysticercosis, a helminthic disease commonly seen in India, Latin America, Eastern Europe and Southern Africa, results from extraintestinal encystation of the larval form of Taenia solium. It is a condition in which man acts as intermediate host instead of definitive host. The most frequent sites of cysticercosis are subcutaneous layers, brain, muscles, heart, liver, lungs, and peritoneum. Oral cysticercosis is considered rare and cause cystic swellings or nodules in the mouth and a precise clinical diagnosis is not usually established. Here, we report a case of oral cysticercosis in a 32-year-old female occurring in the mentalis muscle presenting as asymptomatic nodule.

The master switch: Comparative study of mast cell in oral epithelial dysplasia, oral submucous fibrosis and oral squamous cells carcinoma and their association with inflammation and angiogenesis.

BACKGROUND: Dental and medical practitioners encounter wide spectrum of oral lesions in their day-to-day practice. Many of the lesions such as leukoplakia, oral submucous fibrosis (OSF), etc., are associated with tobacco and betel nut chewing. Oral leukoplakia, OSF, oral lichen planus and oral squamous cell carcinoma (OSCC) are the most commonly occurring oral diseases associated with characteristic clinical and histological features and are associated with chronic inflammation at some stage of the disease process. AIMS: To study and compare the number, morphology and topographical distribution of mast cells in oral epithelial dysplasia (OED), OSF and OSCC and to correlate different types of mast cells with the inflammatory infiltrate and vascularity of the lesions. MATERIALS AND METHODS: Total number of subjects was 120 and equally divided into four groups of 30 as controls, OED, OSF and OSCC cases. Two sections of from each tissue embedded in paraffin wax block were made which were stained with hematoxylin and eosin and toluidine blue stain. Mast cells were counted in five different zones. RESULTS: In the present study, increased numbers of mast cells were seen in all lesions. The cases with mild, moderate and severe inflammation showed increased number of typical (TMCs), atypical (AMCs) and granular mast cells (GMCs), respectively. CONCLUSION: The result of the present study concludes that the mast cells play a key role in mediating the cross links between external angiogenic agent and local immunologic factors.

Mucoepidermoid Carcinoma of Floor of the Mouth - A Rarity.

Mucoepidermoid Carcinoma (MEC) is one of the most commonly occurring malignant salivary gland neoplasm, and contributes 2.8%-15% of all salivary gland tumours. More than half of these cases involve the major salivary glands, primarily the parotid glands and minor salivary glands. Sublingual salivary glands neoplasms are very rare and constitute 0.5% and 1% of all epithelial salivary tumours and approximately 1.5% of the major salivary glands carcinomas. Here we describe a case report of low grade mucoepidermoid carcinoma in the floor of the mouth in a 70-year-old female patient that was mimicking like a ranula clinically.

Gender determination using diagonal measurements of maxillary molar and canine teeth in davangere population.

BACKGROUND: Mesiodistal and buccolingual measurements of the teeth have been the traditional means of determining gender which sometimes are difficult to measure. AIMS AND OBJECTIVES: To assess the degree of sexual dimorphism in upper permanent canines and first molars using diagonal diameters and to evaluate the applicability of diagonal measurements in sex determination. MATERIALS AND METHODS: Mesiobuccal-distolingual (MBDL) and distobuccal-mesiolingual (DBML) measurements of the crown and cervix of maxillary canine and maxillary molar were taken using digital vernier callipers in 100 upper dental casts obtained from 50 males and 50 females of the age group of 17-25 y. RESULTS: With step wise discriminant function analysis, results were statistically significant with males showing higher diagonal measurement values. The classification accuracy for males was 69% and in females was 73% with overall accuracy of 71%. RESULTS showed that canines exhibited highest accuracy with cervical measurements being more dimorphic. MBDL measurements were found to be more reliable. Most dimorphicmeasurements included right molar cervical MBDL, left molar cervical MBDL followed by right and left molar crown MBDL. CONCLUSION: In cases of tooth rotations, crowding, certain fillings and attritions where traditional mesio-distal and bucco-lingual measurements are not applicable, diagonal measurements could be of help in predicting gender.