Exploring miR-155-5p and miR-1246 as Diagnostic and Prognostic Markers in Oral Squamous cell carcinoma.

INTRODUCTION: Oral Squamous Cell Carcinoma (OSCC) is one of the leading cancers worldwide, significantly impacting developing nations. This study aimed to explore the diagnostic and prognostic potential of miR-155-5p and miR-1246 in OSCC in the Indian population, as their comparative roles in this context remain unexplored. MATERIAL AND METHODS: The present cross-sectional study comprised 50 histopathologically confirmed OSCC cases, with adjacent normal mucosa as controls. MiRNA expression was assessed via qRT-PCR and correlated with clinicopathological factors. MiRwalk and miRTargetlink were used for miRNA:mRNA interaction prediction, and gprofiler was employed to analyze validated targets for functional insights. RESULTS: The expression analysis showed a significant upregulation of miR-155-5p and miR-1246 in OSCC tissues compared to adjacent controls. Receiver operating curve analysis revealed that miR-1246 exhibited excellent diagnostic accuracy (AUC = 0.94) compared to miR-155-5p (AUC = 0.69). Higher miRNA levels were associated with age and extracapsular extension while overexpression of miR-1246 was correlated significantly with increased tumor size, tumor grade, TNM staging, and depth of invasion. The analysis for target prediction unveiled a set of validated targets, among which were WNT5A, TP53INP1, STAT3, CTNNB1, PRKAR1A, and NFIB. CONCLUSION: miR-155-5p and miR-1246 may be used as potential prognostic biomarkers in OSCC, with miR-1246 demonstrating superior diagnostic accuracy.

Upregulation of miRNA-196a and miRNA-196b correlates with Bryne's prognostic score in oral squamous cell carcinoma.

BACKGROUND: microRNA(miRNA)-196a and miRNA-196b expression has been found to be dysregulated and involved in tumorigenesis and tumor progression in array of cancers through different targets. The role of these miRNAs together in clinical application is not always consistent and, its prognostic value in oral squamous cell carcinoma (OSCC) is still elusive. This study was performed to investigate the correlation of these miRNAs expression with histological grades of OSCC according to Bryne's histological grading system, to predict prognosis and to evaluate their relationship with clinico-pathological data. METHODS: Real-time quantitative Reverse Transcription Polymerase Chain Reaction (qRT-PCR) was done to evaluate the expression levels of miRNA-196a and miRNA-196b in 75 pairs of OSCC tissue matched with adjacent normal mucosa, used as a control. RESULTS: miRNA-196a and 196b expression in OSCC was significantly higher than that in corresponding adjacent normal tissues (p > 0.001). Also, a significant differential correlation was found in between the expression levels of these two miRNAs (Pearson correlation test r = 0.676, p-value<0.0001). The increased expression of these miRNAs was more frequently observed in OSCC tissues with advanced clinical and pathological TNM stages (IVa and IVb, pIVb respectively, p-value<0.0001). Significant correlation was found between miRNA-196a upregulation and moderate prognostic score (p-value<0.0001) in comparison with good and poor prognostic score of histological grades of OSCC. Sensitivity and specificity for miRNA-196a were 95 % and 85 %, respectively (AUC = 1, 95 % CI = 0.617-0.850; p 0.001), while for miRNA-196b were 94 % and 86 %, respectively (AUC = 0.808, 95 % CI = 0.701-0.916; p0.0001). CONCLUSIONS: These findings suggest that the increased expression of miRNA-196a and 196b may play an important role in tumor progression in OSCC. miRNA-196a might be a useful marker for predicting the clinical outcome of OSCC, especially for advanced stages. In conclusion, our data demonstrate for the first time that these miRNAs may serve as a potent prognostic marker for tumor progression. We further highlight miRNA-196a and miRNA-196b as a promising predictor of prognostic assessment in OSCC.

Mandibular primary intraosseous carcinoma arising from long-standing odontogenic keratocyst.

Primary intraosseous carcinoma (PIOC) of the jaw is a rare neoplasm arising from the lining epithelium of odontogenic cysts or de novo from odontogenic epithelial rests that has no communication with the surrounding mucosa of the upper aerodigestive tract. We present a case of PIOC ex-odontogenic keratocyst (PIOC ex-OKC) in a 35-year-old male. Histopathologic examination revealed a cystic lesion with a fibrous capsule lined by corrugated parakeratinized stratified squamous epithelium resting on a basal cell layer composed of columnar cells exhibiting palisaded hyperchromatic nuclei, features consistent with OKC. Surgical treatment consisted of bilateral crestal and crevicular incision, a reflection of the flap, breaking of all OKC locules, creation of a continuous cavity, and fitting of a decompression mold around the mandibular teeth. This case highlights the importance of knowing the features of PIOC and considering PIOC in the differential diagnosis of malignant tumors of odontogenic epithelium for timely surgical treatment.

Evaluation of Diagnostic Significance of Salivary miRNA-184 and miRNA-21 in Oral Squamous Cell Carcinoma and Oral Potentially Malignant Disorders.

BACKGROUND: Altered levels of miRNAs might affect the pathogenesis of oral squamous cell carcinoma (OSCC) and oral potentially malignant disorders (OPMD). This study evaluated the diagnostic potential of salivary miRNA-21 and miRNA-184 in OSCC and OPMD. METHODS: We recruited a total of 90 subjects including OSCC, OPMD, and healthy controls. RNA was isolated from the saliva samples of the study subjects. Expression of miRNA-21 and miRNA-184 was analyzed using qRT-PCR. Their levels were compared and the diagnostic cut-off was determined using the ROC curve. RESULTS: There was a significant increase in miRNA-21 and a decrease in miRNA-184 in OSCC and OPMD as compared to healthy controls (p < 0.001). Levels of salivary miRNA-21 and miRNA-184 can differentiate OSCC and OPMD from controls and premalignant conditions from malignant conditions. CONCLUSION: Salivary miRNA-21 and miRNA-184 may be beneficial for the early detection of OSCC and OPMD. Also, saliva can be used for detecting neoplastic transformation of oral mucosa since it is non-invasive and easily accessible.

Lifting the lid over the pearl: A histological insight.

Epithelial pearls and Keratin pearls are pathognomonic of squamous cell carcinoma. However, their histogenesis is not well understood. Only a handful of studies have been conducted in the past in this regard. This brief communication aims to understand the formation of these pearls with a few of our own experiences.

Epidermoid cysts of the orofacial region: A clinico-pathological study of 13 cases with review of literature.

Background: Epidermoid cysts (ECs) are uncommon benign cystic lesions derived from the germinative epithelium. Head and neck ECs constitute only 7% of all ECs whereas only 1.6% are seen intraorally. The floor of the mouth is the commonest intraoral site whereas tongue, lips, buccal mucosa, and jaws are less commonly involved intraoral sites. To date, very few large case series of ECs of head and neck have been published. To the best of our knowledge, this is the third-largest case series of 11 intraoral ECs along with 2 extra-oral cases in the pre-auricular region. Aims: To highlight the typical and atypical features of ECs in the common as well as rare sites and draw attention to its consideration as a differential diagnosis for head and neck masses. Settings and Design: Archival data of 13 histopathological cases identified as ECs were analyzed from the Department of Oral Pathology at a tertiary dental hospital and college in New Delhi from 2007 to 2020. Materials and Methods: The demographic, clinical, radiographic, histopathological features, and treatment modalities were recorded and analyzed. Statistical Analysis Used: Appropriate statistical tests were used. Results: The study found strong male predilection in the ratio of 10:3 with an average age of presentation as 28 years. The pre-auricular region and floor of the mouth were the common sites involved followed by buccal mucosa, lips, and jaws. All patients presented with slowly growing swelling with dysphagia, dyspnea, and dysphonia seen in larger cysts on the floor of the mouth. Microscopically, all cases were lined with stratified squamous epithelium filled with laminated layers of keratin. Two cases showed the presence of melanin. One case showed recurrence even after complete surgical excision. Conclusion: ECs, though a rare entity, should be considered in differential diagnosis for head and neck masses and require close follow-up due to their potential for malignant transformation.

Cysticercosis: Unearthing the worm.

Cysticercosis, a helminthic disease caused by T. Solium, is a major health concern in developing and underdeveloped nations of the world. If left untreated, it may lead to severe neurological and ophthalmic complications. The diagnosis of oral cysticercosis depends on the identification of the larva in the biopsied tissue. However, an accurate diagnosis can be challenging, if the larva is dead because of which it cannot be identified. In such a scenario, step by step approach to unearth the worm is discussed here.

Potential role of mast cells and eosinophils in establishing the clinical severity of oral lichen planus.

BACKGROUND: Oral lichen planus (OLP) can be broadly classified clinically as an erosive (e) and non-erosive (ne) form. Mast cells (MCs) are granules containing cells that play an important role in the etiopathogenesis of OLP and have bidirectional interaction with eosinophil. In the present study, MCs and eosinophil densities were studied to find out if they could aid in the clinical severity of eOLP and neOLP. METHODS: The study group included 20 cases of eOLP and 30 cases of neOLP. All the sections were stained with special stains, toluidine blue (TB) for MCs and Congo red (CR) for eosinophils. Histopathological analysis was done by using Nikon NIS Elements Version 5.30 software (Nikon, Tokyo, Japan) by applying 100×100 µm grids to standardize the field of evaluation. Three subepithelial zones were demarcated as follows: 1) zone I was up to a subepithelial depth of 100 µm from the basement membrane; 2) zone II was beneath zone I from 100 µm to 200 µm; and 3) zone III was beneath zone II from 200 µm to 300 µm. In each of these zones, five high power fields with high cellular density were chosen to count MCs and eosinophils. RESULTS: The results of the study showed a significant increase in the number of MCs (intact and degranulated) in zone II and III compared to zone I in both eOLP and neOLP. Eosinophil density was significantly higher in zone II of both eOLP and neOLP when compared to zone I and III. We also observed a significantly higher number of both MCs and eosinophils in eOLP compared to neOLP. This difference was most prominent between zone II of eOLP and neOLP. No significant difference between granulated and degranulated MCs was found between the three zones. CONCLUSIONS: Increased MC and eosinophil densities, as well as their interactions in eOLP, suggest that they play a significant role in etiopathogenesis and in establishing the clinical severity of the disease.

Bilateral Dentigerous Cysts of the Permanent Mandibular First Molars Treated with Marsupialization.

Bilateral dentigerous cysts (DC) associated with unerupted mandibular first molars in a non-syndromic pediatric individual are rare. Secondary infections may lead to complications, such as discomfort due to pain, disfigurement caused by enlargement of the cyst with cortical expansion of the jawbone, displacement of teeth and paraesthesia of the adjacent nerve. This case report describes the occurrence of bilateral DC in an eight-year-old patient. Marsupialization was the treatment of choice to preserve the permanent teeth and other adjacent tissues.

"Is it time for personalised medicine for Ameloblastoma?": A hypothesis.

Ameloblastoma is a benign odontogenic tumor that is locally destructive. The most common treatment option is surgery, which often results in disfigurement of the face. BRAFV600E is the common gene mutation associated with its pathogenesis. Therefore, this paper hypothesizes the use of targeted drug therapy against this mutated gene.

Dentinogenic Ghost Cell Tumor associated with Odontoma: A unique Histopathological Entity and its Surgical Management.

We report a rare case of dentinogenic ghost cell tumor (DGCT) associated with complex composite odontoma in a 17 years male affecting the posterior segment of the mandible. On radiographic examination, there was a well-defined multilocular radiolucency surrounding the radio opaque mass with respect to 44, 45 and 46. Histopathologically it showed ameloblastomatous proliferation with dentin like areas and ghost cells. It was associated with tooth like structures consisting of dentin, cementum and pulp like areas. DGCT with odontoma is extremely rare with only two cases being reported in literature till date. The management with its rare occurrence is discussed here.

Ghost cells unveiled: A comprehensive review.

BACKGROUND: Ghost cells (GCs) are cells with distinct intracytoplasmic keratinization, which leads to the preservation of the cellular outline with a clear area corresponding to the previous nucleus location. GCs may show various patterns, such as degeneration, tissue granulation, and calcification. Their true nature and the mechanism regulating the conversion of odontogenic epithelial cells into GCs remain unclear. GC keratinization is different from normal keratinization as they are larger than keratotic squames, are frequently vacuolated, and have prominent nuclear membrane remnants. Few cystic lesions, odontogenic tumors, and non-odontogenic tumors, such as calcifying odontogenic cyst, craniopharyngioma, pilomatrixoma, odontoma, dentinogenic ghost cell tumor, and ghost cell odontogenic carcinoma, exhibit GCs as a typical feature. The Wnt and Notch signaling pathways play a role in the histogenesis of the neoplasms. HIGHLIGHT: The review clarifies the various proposed hypotheses of the histogenesis of GCs, including molecular pathogenesis. Diagnostic workup for the identification of GCs, including special staining and immunohistochemistry, has been extensively discussed. A stepwise algorithm for identifying odontogenic and non-odontogenic lesions containing GCs has been proposed. Additionally, the prognostic role of GCs in the lesions has been elucidated. CONCLUSION: Among the various hypotheses of the origin of GCs, we suggest that aberrant keratinization is the most accepted based on various immunohistochemical studies and special staining characteristics. GCs are a distinct characteristic entity of many odontogenic and non-odontogenic lesions; however, it remains controversial whether their presence has any pathognomonic role in the biological nature of these lesions.

Preoperative platelet-lymphocyte ratio and neutrophil-lymphocyte ratio as predictors of occult lymph node metastasis detected using Desmoglein 3 and Cytokeratin in Indian population.

Aim: This study aims to assess whether preoperative platelet-lymphocyte ratio (PLR) and neutrophil-lymphocyte ratio (NLR) can predict occult metastasis in oral squamous cell carcinoma (OSCC). Materials and Methods: Thirty-five OSCC cases were analyzed for clinicopathological and hematological data. Cases without metastasis (pN0) were checked for micrometastasis immunohistochemically using Desmoglein 3 (DSG3) and Cytokeratin (CK). Mean PLR and NLR were compared and analyzed between the study groups. Results: Metastatic deposits were detected in 9 out of 26 pN0 cases (34.6%) accounting for 11 out of 62 (17%) lymph nodes subjected to immunohistochemistry. The mean PLR was higher in OSCC cases with or without occult metastasis in comparison to controls (P < 0.001). No significant difference was found in the mean PLR and NLR between OSCC cases with and without occult metastasis. Furthermore, we found DSG3+ sinus histiocytes within the lymph nodes in majority of cases which is least reported in literature. Conclusion: A significant percentage of cases showed occult metastasis in this study which led to upstaging of tumor. Although PLR was elevated in OSCC cases, it did not have a positive correlation with the presence of occult metastasis but was able to successfully distinguish OSCC patients from healthy individuals.

Evaluation of Endosequence Root Repair Material and Endocem MTA as direct pulp capping agents: An in vivo study.

The study aimed to assess the human pulpal response to direct pulp capping using Endosequence Root Repair Material (ERRM) and Endocem MTA against ProRoot MTA as control. Intentional direct pulp capping was done using the three materials in 30 caries-free human premolars. After 30 days, the teeth were extracted, fixed and decalcified. Sections were prepared for histologic examination using light microscopy. Thickness and quality of dentine bridge formed and level of inflammation were evaluated. Difference between dentine bridge thickness of the three groups was statistically significant. Mean thickness of dentine bridges formed in ProRoot MTA group was greater than other two groups, and difference was statistically significant. Difference in dentine bridge thickness between ERRM and Endocem MTA was statistically significant. No significant difference was observed between the 3 groups with respect to inflammation. Results indicate ProRoot MTA performed best with ERRM giving better results than Endocem MTA.

Does Loss of Immunohistochemical Expression of Glypican 3 in Oral Squamous Cell Carcinoma Play a Role in the Wnt/ß-catenin Signaling Pathway?

Glypican 3 (GPC3) is a cell membrane protein and plays a dual role, as a tumor suppressor and oncogene, depending on its structure. It is known to regulate the Wnt/ß-catenin signaling pathway and affect cell growth and proliferation. ß-catenin plays a major oncogenic role in progression of oral squamous cell carcinoma (OSCC); thus, this study aimed to explore the relationship between ß-catenin and GPC3 in OSCC. Immunoexpression of GPC3 and ß-catenin was evaluated semiquantitatively in tumor tissue (n=80) and normal oral mucosa tissue (n=20). For GPC3, the percentage of stained cells and the staining intensity were assessed. For ß-catenin, the percentage of stained cells, localization, and intensity of staining were assessed at the tumor-invasive front. The Pearson correlation was used to determine the correlation between the GPC3 and ß-catenin immunoreactivity. Significantly decreased expression of GPC3 (P=0.008) and a highly significant difference in the case of localization of ß-catenin (P=0.0001) were observed in OSCC when compared with normal oral mucosa. Cytoplasmic expression with a shift of ß-catenin expression to the nucleus was seen in OSCC in comparison with primarily membranous and membranous and cytoplasmic staining in normal mucosa. A significant difference was observed with respect to localization of stain, with ß-catenin staining moving to the nuclear compartment with an increase in the tumor grade (P=0.011). No correlation was observed between ß-catenin and GPC3 expression in OSCC cases. It is concluded that loss of expression of GPC3 in OSCC compared with normal oral mucosa indicates that it plays the role of a tumor suppressor gene in OSCC and its expression is therefore silenced in OSCC.

Chorea-acanthocytosis: A Case Report with Review of Oral Manifestations.

Chorea-acanthocytosis (ChAc) is an autosomal recessive, progressive neurological disorder due to mutation in VPS13A gene causing defects in sorting of protein making the cell membrane unstable, leading to star-shaped erythrocytes. This neurological disorder includes features such as elevated creatinine kinase, atrophy of basal ganglia, and oral manifestations such as frequent cheek and tongue biting. It is a rare neurological condition with an estimate of <1000 cases worldwide. A case of 47-year-old male patient with a history of seizures and neurological problems presenting with oral ulceration has been discussed. The diagnosis of ChAc was confirmed by molecular investigations showing VPS13A gene mutation. The physical appearance includes chorea and dystonia with impaired gait. We attempt to highlight the oral features of ChAc. The oral manifestations include frequent tongue and cheek biting occurring due to dystonia affecting the muscles of head and neck region.

Significance of HIF-1α Expression and LOXL-2 Localization in Progression of Oral Squamous Cell Carcinoma.

Backgroud: In the microenvironment of Oral Squamous Cell Carcinoma (OSCC), Hypoxia-inducible transcription factor 1 (HIF-1) is a very important chemical mediator in the microenvironment of OSCC through which cells respond to hypoxia. LOXL-2 participates in ECM remodelling, and also in regulating epithelial-to-mesenchymal transition, epithelial cell polarity and differentiation. Aim/material and methods: The present study was conducted on 90 histopathologically proven cases of OSCC to ascertain the role of HIF-1α and LOXL-2 in OSCC. Immunoexpression of both HIF-1α and LOXL-2 was analyzed both quantitatively and qualitatively and compared with tumor stage, nodal stage, clinical stage, and histological grade. Results: Tumor stages and nodal stages had significant correlation with HIF-1α expression and localization of LOXL-2 immunoexpression respectively. Conclusion: This is probably the first study to analyze LOXL-2 localization in OSCC. Alteration in the immunoexpression of LOXL-2 from nuclear to cytoplasmic and HIF-1α immunoexpression might be an important factor in progression of OSCC.

Ghost cell characterization in calcifying odontogenic cysts and dentinogenic ghost cell tumors: An immunohistochemical study.

OBJECTIVES: The aim of our study was to ascertain the true nature of ghost cells (GCs) by immunolocalization of cytokeratin (CK) 6, CK19, and amelogenin in calcifying odontogenic cysts (COCs) and dentinogenic ghost cell tumors (DGCTs) in an attempt to determine the nature of this unique cell. METHODS: A total of thirteen cases (six COCs and seven DGCTs) were examined immunohistochemically, in order to compare immunoreactivity for CK6, CK19, and amelogenin in odontogenic GCs. RESULTS: Positive expression of amelogenin (92.3%) and CK6 (77%) was chiefly found in GCs. CK19 expression was observed in the cytoplasm of odontogenic epithelial cells of the lining epithelium. GCs were devoid of CK19 expression and were positive only on the cytoplasmic periphery. CONCLUSION: In the current study, GCs showed accumulation of amelogenin and hard keratins in their cytoplasm during pathological transformation.

A novel approach for extracting DNA from formalin-fixed paraffin-embedded tissue using microwave.

BACKGROUND: Formalin-fixed paraffin-embedded (FFPE) tissue is the gold standard procedure for tissue preservation. However, the extraction of DNA is a cumbersome procedure as the extracted DNA is fragmented because of the cross-linking effect of formalin. Hence, the aim of the present study is to extract DNA from FFPE tissues using different techniques with a specific objective of comparing the extracted DNA, both quantitatively and qualitatively. METHOD: Ten samples of FFPE tissues were retrieved from the archives of the Department of Oral Pathology. Total genomic DNA was extracted by different methods which included QIAamp DNA FFPE Tissue Kit, Norgen DNA FFPE Tissue Kit, phenol-chloroform method, mineral oil-based extraction, M/10 NaOH solution method, and microwave method. A 280-base pair sequence was selected for evaluation of downstream amplification. RESULTS: The statistical analysis was performed using unpaired student's t-test to compare the DNA yields and quality obtained by microwave methods with other methods using SPSS software. Total genomic DNA retrieved by the microwave method was superior to other methods both qualitatively and quantitatively. CONCLUSION: DNA extraction from FFPE tissues is an onerous task as irreversible bonds form between the nucleic acid during fixation which are difficult to break during DNA retrieval. Hence, the microwave method provides good total genomic DNA which gives better downstream results when compared with other methods.