Role of MMP1 and MMP10 in local invasion and distant metastasis in different levels of oral squamous cell carcinoma - A immunohistochemical comparative study.

Background: Matrix metalloproteinases (MMPs) are basically a part of a large family of proteolytic enzymes. They play an important role in degrading extracellular matrix and basement membrane, which is a basic mechanism in local invasion and tumour metastasis. The aim of this study was to evaluate immunohistochemically the expression of MMP1 and MMP10 in tumour invasion locally and at distant levels, including lymph nodes at different levels in oral squamous cell carcinoma (OSCC). Materials and Methods: A total of 50 tissue samples with clinically confirmed OSCC and 15 normal oral mucosal tissues will be included in the study. Immunohistochemical staining will be performed for the demonstration of MMP1 and MMP10 in lesional tissue, perilesional tissue, and lymph nodes of different levels that were evaluated with respect to microscopic features. Results: All OSCC cases had MMP1 and MMP10 expression levels. The expression increased as the nodal level increased from level I to level V. This difference was statistically significant at P < 0.001 Both MMPs were not expressed in normal epithelial cells. There was no significant correlation between MMP1 and MMP10 expression. Conclusion: This study showed that MMP1 and MMP10 are expressed in the tissues of OSCC and may serve as prognostic indicators for the disease.

Role of hypoxia and epithelial-mesenchymal transition in the formation and maintenance of oral cancer stem cells in oral squamous cell carcinomas and metastatic lymph node: An immunohistochemical analysis.

Background: Inspite of having advanced treatment modalities the overall survival rate in oral squamous cell carcinoma (OSCC) remains poor. This is considered to be mainly due to local recurrence and distant metastasis. Various studies have concentrated on the role of oral cancer stem cells (OCSCs) in the progression and metastasis of OSCC. However, the role of tumor microenvironment components has been less delved into. Hence clarity on cell biology and metastatic potential OCSCs is essential for the development of more effective anti-cancer treatment. Aim: To establish the role of OCSCs in different grades of OSCC and metastatic lymph nodes through the expression of cluster of differentiation 44 (CD44). To demonstrate and correlate the role of hypoxia and Epithelial mesenchymal transition (EMT) in the various grades and metastatic lymph nodes in the formation and maintenance of OCSCs by employing Hypoxia-inducible factor-1 Alpha (HIF 1α) and Snail respectively. Method and Material: A total of 36 cases of OSCC, 12 from each grade and 12 normal oral mucosal tissues were included in the study. Immunohistochemical staining was performed for the demonstration of CD44, HIF1α, and Snail. Statistics: Descriptive analysis, Chi-square, and Spearman's rank correlation were used to analyze frequency and proportion, to compare expression and correlate between lesion proper and lymph node in each group respectively. Results: Significant expression of CD44, HIF1 α, and Snail among advancing grades of OSCC and their metastatic lymph node were observed. A positive correlation was seen between them. Conclusions: The prognosis of OSCC can be improved by better understanding and targeting the molecules involved in the formation and maintenance of OCSCs.

Molecular characterization and antifungal activity of lipopeptides produced from Bacillus subtilis against plant fungal pathogen Alternaria alternata.

Over 380 host plant species have been known to develop leaf spots as a result of the fungus Alternaria alternata. It is an aspiring pathogen that affects a variety of hosts and causes rots, blights, and leaf spots on different plant sections. In this investigation, the lipopeptides from the B. subtilis strains T3, T4, T5, and T6 were evaluated for their antifungal activities. In the genomic DNA, iturin, surfactin, and fengycin genes were found recovered from B. subtilis bacterium by PCR amplification. From different B. subtilis strains, antifungal Lipopeptides were extracted, identified by HPLC, and quantified with values for T3 (24 g/ml), T4 (32 g/ml), T5 (28 g/ml), and T6 (18 g/ml). To test the antifungal activity, the isolated lipopeptides from the B. subtilis T3, T4, T5, and T6 strains were applied to Alternaria alternata at a concentration of 10 g/ml. Lipopeptides were found to suppress Alternaria alternata at rates of T3 (75.14%), T4 (75.93%), T5 (80.40%), and T6 (85.88%). The T6 strain outperformed the other three by having the highest antifungal activity against Alternaria alternata (85.88%).

COVID-19 Associated Mucormycosis (CAM): A Single Hospital-Based Study.

Background: Opportunistic fungal infections like Mucormycosis in Coronavirus Disease 2019 (COVID-19) patients have posed a great challenge to health care professionals, especially in developing countries like India. Hence, there is a need to understand the biological behaviour of COVID-19 associated Mucormycosis (CAM) to establish standard treatment Protocols and to reduce mortality. Aims: This study aims is to assess the type of Mucormycosis among COVID-19 patients in study population and compare the findings with clinical, radiological and haematological parameters along with treatment and surgical management. Methods and Material: This retrospective, observational study included 60 cases of CAM reported to the Department of Oral and Maxillofacial Surgery at the tertiary care centre, Karnataka Institute of Medical Sciences, Hubli. Data about various parameters were tabulated and analysed statistically. Statistical Analysis Used: Bivariate analysis was done using the Chi-Square test to assess the relationship between the type of Mucormycosis and other variables. Spearman's Correlation test was used to assess the correlation between types of Mucormycosis with the other variables. Linear regression analysis was performed to assess the response variable related to the type of Mucormycosis. Results: About 50% of subjects presented with "Rhino orbital" type of Mucormycosis. Palatal discoloration and palatal erosion was the most common oral manifestation among "only Sinus" and "Rhino orbital" types of Mucormycosis (P = 0.00). Significant association (P = 0.29) was found between the type of Diabetes mellitus and Mucormycosis. Conclusions: The study indicates that DM is the most commonly associated comorbidity in CAM patients. Hence, a thorough understanding of the underlying comorbidity and its close monitoring during and after COVID-19 infection is mandatory for successful treatment outcomes.

Senescent Fibroblast in Oral Submucous Fibrosis Aids in Disease Progression and Malignant Transformation.

Aim: To study the role of senescent fibroblasts (SFs) and its secretory phenotypes promoting fibrosis and malignancy in oral submucous fibrosis (OSMF). Material Methods: A total of 20 cases of OSMF and 20 normal oral mucosal tissues were taken for the study. The tissue sections were stained for IHC-immunohistochemistry with senescent cell marker p16INK4a. The supernatant solution of the transport medium was studied for leached out senescent associated secretory proteins (SASP): matrix metalloproteinases 9 (MMP9), interleukins 6 (IL6), vascular endothelial growth factor (VEGF). Tissues were studied for malignant transformation with p53 and Ki67. Fibrosis in the OSMF was tested with lysyloxidase (LOX). The Statistical Package for the Social Sciences (SSPS) statistical software was used to analyze the data. Results: IHC staining for p16INK4ashowed positivity in the connective tissue of OSMF cases which was statistically significant. Antibody assay using enzyme-linked immunosorbent assay (ELISA) showed elevated levels of secretoproteins IL6, MMP9, VEGF in OSMF cases. LOX enzyme levels were also significantly increased in OSMF cases. Proliferative markers Ki67 and p53 were positive in IHC staining in the epithelium of OSMF. Conclusion: This study confirms the presence of SF and its secreto phenotypes in OSMF and showed increased LOX expression which is implicated in fibrosis. These findings suggest that SF may contribute to fibrosis in OSMF. The study also confirms the malignant transformation of the overlying epithelium as shown by p53 and Ki67 positivity.

A Comparative Immunohistochemical Study of Presence and Distribution Pattern of Stromal Myofibroblast in Oral Dysplasia and in Different Grades of Oral Squamous Cell Carcinoma.

AIMS AND OBJECTIVES: Squamous cell carcinoma (SCC) is a common oral malignancy with a poor survival rate. Early tumorigenesis is marked by transdifferentiation of fibroblasts to myofibroblasts (MFs), which is supported by growth factors and cytokines expressed by tumor cells. The expression of alpha-smooth muscle actin (αSMA) marker correlates with the activation of MFs. This study was undertaken to compare the frequency and distribution of αSMA immunoexpression in oral epithelial dysplasia (OED) and OSCC. MATERIALS AND METHODS: This study was conducted on samples collected from patients with oral epithelial dysplasia and oral SCC who visited Rajarajeswari Dental College and Hospital, Bengaluru. Tissue sections were subjected to Immunohistochemistry using αSMA marker, and cells were counted. The obtained data was subjected to Kruskal-Wallis test and Mann-Whitney U-test. RESULTS: On performing Kruskal-Wallis test and Mann-Whitney U-test between the three groups (normal oral mucosa, OED, and OSCC) statistically significant result was found in the frequency between OED and OSCC and between normal tissue and OSCC. On comparing the distribution pattern, statistically significant result was found between OED and OSCC and between normal tissue and OSCC. CONCLUSION: The expression of MFs increases as the disease progresses from high-grade epithelial dysplasia to invasive OSCC. Poorly differentiated SCC showed more attendance of positive MFs in the stroma than other grades of OSCC. The rise in the number of αSMA-positive MFs and change in distribution pattern in OSCC can be associated with tumor invasive characteristics. Thus, the proliferation of MFs may be used as a stromal marker of premalignancy and malignancy.

Forensic photography: Prospect through the lens.

Forensic photography is an indispensable tool in modern forensic odontological protocol which aids in investigative procedures, maintenance of archival data, and to provide evidence that can supplement medico legal issues in court. Proper selection and implementation of the appropriate photography and computer equipment combined with necessary training and correct workflow patterns make incorporating photography into the field of forensics, an easily obtainable goal. The role of the forensic photographer is crucial, as a good skill in photography with updated knowledge of the mechanics and techniques involved is required for proper documentation of evidence. This paper aims to shed light on the various aspects of forensic photography with emphasis on its diverse applications and advancements.

Rapid tissue processing technique: A novel method using methyl salicylate.

INTRODUCTION: Tissue processing involves transition of the biopsy tissue in graded concentration of various chemicals to make the tissue amiable for sectioning. The entire process takes 2-3 working days before a microscopic slide is ready for diagnosis. In order to shorten the turnaround time, rapid tissue processing method using methyl salicylate was developed. AIM: The aim of this study is to develop a rapid tissue processing technique using methyl salicylate as a clearing agent and to compare it with routine tissue processing technique. MATERIALS AND METHODS: A total of 70 tissue specimens were cut into two equal halves. One each was processed by routine processing technique (RoPT) and rapid processing technique (RaPT). Tissue specimens were measured before and after processing. Quality of staining and cellular-level shrinkage were observed and scored for specimens. Statistical analysis using Welch's unequal variances t-test was performed. Costs of chemicals in both the techniques were compared to see the cost-effectiveness of RaPT. RESULTS AND CONCLUSION: Outcomes of both the processing techniques were comparable with statistically not significant P values for all the parameters. Hence, the results of RaPT technique are satisfactory, and the use of this technique may prove beneficial to pathology laboratories.

Histopathological Evaluation of Dental Follicle Associated with Radiographically Normal Impacted Mandibular Third Molars.

INTRODUCTION: Removal of impacted mandibular third molar is a common procedure performed in oral surgery. Indications for removal of the third molar have generated much discussion in dentistry. The presence of pericoronal pathosis is generally accepted reason for the extraction of impacted mandibular third molars. Radiographic pathology is usually defined as a pericoronal radiolucency measuring about 2.5 mm or larger in any dimension. PURPOSE: This study aims to evaluate the histopathologic changes in radiographically normal dental follicles associated with impacted mandibular third molars. METHODS: After extraction of 50 impacted mandibular third molars, dental follicle associated with extracted teeth was placed in 10% formalin solution. Histopathologic examination was done. The type of pathological changes was recorded based on histopathological reports. Patients' gender, age, and radiographic parameters were recorded. RESULTS: Cystic changes were found in 24% of radiographically normal impacted mandibular third molars. Significant cystic changes were observed in position A vertically impacted lower third molars (ILTMs). Male predominance for cystic change was seen with a male-female ratio of 2:1. CONCLUSION: The result of the present study shows that radiographic analysis may not be reliable technique for the diagnosis of cystic changes in ILTM. The presence of cystic changes can be found even in radiographically normal impacted mandibular third molars. Hence, we recommend that impacted mandibular third molars should be removed and histopathological analysis should be conducted on all surgically extracted dental follicles.

Gender determination by odontometric method.

CONTEXT: Gender determination is central in establishing personal identification from human skeletal remains. The study was conducted to find out the accuracy with which gender can be determined by odontometric methods. AIMS: To investigate the mesiodistal (MD) and buccolingual (BL) dimensions of all the teeth of permanent dentition to find new parameters to differentiate between male and female teeth and to assess whether each type of linear measurement can be used independently in odontometric sex differentiation. MATERIALS AND METHODS: The study was conducted at a dental college on a composite group of 500 individuals comprising 250 males and 250 females. Impressions of upper and lower jaws were made with alginate impression material and casts prepared with dental stone. A digital Vernier calliper was used to measure the BL and MD dimensions of all the upper teeth except the third molars. STATISTICAL ANALYSIS USED: The results were subjected to statistical analysis using univariate analysis and linear stepwise discriminant function analysis to find the variables which discriminate gender significantly. RESULTS: The MD and BL dimensions between males and females were statistically significant. The predicted value for correct classification of gender was also statistically significant. CONCLUSIONS: The ability to differentiate gender in the population using stepwise discriminant functions was found to be very high with 99.8% accuracy with males showing statistically larger teeth than females. This is similar to the near 100% success in gender determination using pelvic and skull bones.

Concurrent central odontogenic fibroma and dentigerous cyst in the maxilla: A rare case report.

Central odontogenic fibroma (COF) represents a rare group of benign odontogenic tumor, while dentigerous cyst (DC), on the other hand, is the odontogenic cyst that encloses the crown of an unerupted tooth. A combination of COF and DC is rare and not reported in literature till date. The aim of this paper is to report a case of COF occurring concurrently with DC. The simultaneous occurrence of COF and DC raises the question of whether it is a collision tumor or their simultaneous occurrence is just a coincidence. In our case, the probable cause of simultaneous occurrence might be that COF would have displaced 23 resulting in the formation of DC.

Stereomicroscopic study on unsectioned extracted teeth.

INTRODUCTION: Age has been considered as a reliable marker for establishing the identity of a person in the field of forensic medicine. Teeth are useful skeletal indicators of age at death since it can survive for decades. Nondestructive methods ensure the evident preservation of dental hard tissues that reflect age changes from the cradle to the grave. Therefore, an attempt was made for estimating the age using the nondestructive method. AIMS AND OBJECTIVES: The aim of this study to assess whether physiological changes of the teeth allow possible correlation for accurate age estimation and to establish a graduation standard by microscopic observation for a better age correlation. MATERIALS AND METHODS: The study was carried on 209 teeth samples extracted for orthodontic treatment or periodontal diseases comprised both maxillary and mandibular teeth across different age groups. The assessment of these changes was carried out by well-established standard methods with some proposed modifications. RESULTS: Pearson correlation analyses revealed root dentin translucency with the highest correlation (r = 0.97) followed by periodontal ligament attachment (r = 0.95), root dentin color (r = 0.95), and attrition being the least correlated (r = 0.90). All the parameters taken for the study contributed to stepwise linear regression analysis (R = 0.98; P < 0.01) indicating a strongly positive relationship between age and the changes observed. A regression formula was obtained with mean error age difference ±1.0 years. CONCLUSION: The present study showed that extracted tooth is highly significant in identifying the age without being sectioned or further processed and also signifies the use of microscope for observation of these changes, thus reducing the errors of calibrating the age.

Neural crest: The fourth germ layer.

The neural crest cells (NCCs), a transient group of cells that emerges from the dorsal aspect of the neural tube during early vertebrate development has been a fascinating group of cells because of its multipotency, long range migration through embryo and its capacity to generate a prodigious number of differentiated cell types. For these reasons, although derived from the ectoderm, the neural crest (NC) has been called the fourth germ layer. The non neural ectoderm, the neural plate and the underlying mesoderm are needed for the induction and formation of NC cells. Once formed, NC cells start migrating as a wave of cells, moving away from the neuroepithelium and quickly splitting into distinct streams. These migrating NCCs home in to different regions and give rise to plethora of tissues. Umpteen number of signaling molecules are essential for formation, epithelial mesenchymal transition, delamination, migration and localization of NCC. Authors believe that a clear understanding of steps and signals involved in NC formation, migration, etc., may help in understanding the pathogenesis behind cancer metastasis and many other diseases. Hence, we have taken this review to discuss the various aspects of the NC cells.

Pseudoepitheliomatous Hyperplasia in Oral Lesions: A Review.

Pseudoepitheliomatous hyperplasia (PEH) is a histopathological reaction pattern to various stimuli, which includes trauma, infection, inflammation, neoplasia. It is seen as tongue like epithelial proliferation invading the connective tissue and should not be mistaken for squamous cell carcinoma (SCC). This review enlists oral lesions which exhibit PEH with a note on how to differentiate SCC from PEH.

Age estimation using annulations in root cementum of human teeth: A comparison between longitudinal and cross sections.

BACKGROUND: Age estimation is an important factor in the identification of an individual in forensic science. Research indicates that cemental annulations may be used more reliably than other morphological or histological traits of human skeleton for age estimation. MATERIALS AND METHODS: Twenty-five teeth were sectioned longitudinally, and twenty-five teeth were cross-sectioned at the mid portion of the root. Sections were ground, mounted and viewed under a bright light microscope. The area selected for counting was photographed under ×10 objective, magnified 5 times; cemental lines were counted and added to the eruption age of that patient, to obtain the chronological age. The statistical software SAS 9.2, SPSS 15.0, Stata 10.1, MedCalc 9.0.1, Systat 12.0 and R environment ver.2.11.1 were used for the analysis of the data. RESULTS: The P value comparing actual age and calculated age using longitudinal sections is moderately significant and the P value comparing actual age and calculated age in the age group of <30 years is significant. INTERPRETATION AND CONCLUSION: The middle third of tooth root was most suitable to count annulations. The cross sections are easier to count but longitudinal sections give more appropriate results on age estimation. Though the procedure predicts under assessment of age in the younger age group and over assessment of age in the older age group, it provides a close estimate of the actual age of an individual. It can be correlated with other age estimation methods for better reliability.

Risk of tumor cell seeding through biopsy and aspiration cytology.

Cancer cells, besides reproducing uncontrollably, lose cohesiveness and orderliness of normal tissue, invade and get detached from the primary tumor to travel and set up colonies elsewhere. Dislodging neoplastically altered cells from a tumor during biopsy or surgical intervention or during simple procedure like needle aspiration is a possibility because they lack cohesiveness, and they attain the capacity to migrate and colonize. Considering the fact that, every tumor cell, is bathed in interstitial fluid, which drains into the lymphatic system and has an individualized arterial blood supply and venous drainage like any other normal cell in our body, inserting a needle or a knife into a tumor, there is a jeopardy of dislodging a loose tumor cell into either the circulation or into the tissue fluid. Tumor cells are easier to dislodge due to lower cell-to-cell adhesion. This theory with the possibility of seeding of tumor cells is supported by several case studies that have shown that after diagnostic biopsy of a tumor, many patients developed cancer at multiple sites and showed the presence of circulating cancer cells in the blood stream on examination. In this review, we evaluate the risk of exposure to seeding of tumor cells by biopsy and aspiration cytology and provide some suggested practices to prevent tumor cell seeding.

Expression of CD34 and CD68 in peripheral giant cell granuloma and central giant cell granuloma: An immunohistochemical analysis.

BACKGROUND: Central and Peripheral giant cell granulomas of jaws are uncommon, benign, reactive disorders that are characterized by the presence of numerous multinucleated giant cells and mononuclear cells within a stroma. The origin of the multinucleated giant cells is controversial; probably originating from fusion of histiocytes, endothelial cells and fibroblasts. OBJECTIVE: To assess the expression of CD34 and CD68 in central and peripheral giant cell granulomas to understand the origin of these multinucleated giant cells. MATERIALS AND METHODS: Twenty cases of Central and Peripheral giant cell granulomas were evaluated immunohistochemically for CD34 and CD68 proteins expression. RESULTS: Immunopositivity for CD34 was seen only in cytoplasm of endothelial cells of blood vessels; whereas, consistent cytoplasmic immunopositivity for CD68 was seen in few stromal cells. Statistical significance was seen in mean number of multinucleated giant cells, mean number of nuclei in multinucleated giant cells, CD68 expression and ratio of macrophages to multinucleated giant cells among two lesions. CONCLUSION: Although the central giant cell granulomas share some clinical and histopathological similarities with peripheral giant cell granulomas, differences in mean number of nuclei in multinucleated giant cells and CD68 immunoreactivity may underlie the distinct clinical behavior.