Leptin levels in gingival crevicular fluid during canine retraction: in vivo comparative study.

OBJECTIVE: To evaluate and compare the leptin levels in gingival crevicular fluid (GCF) and rate of canine retraction using an elastomeric chain and nickel-titanium (NiTi) closed coil spring. DESIGN: In vivo, clinical study. SETTING: Orthodontic department at ITS Centre for Dental Studies and Research, Ghaziabad, India. PARTICIPANTS: Twenty-seven patients reported to the Department of Orthodontics at ITS Centre for Dental Studies and Research. METHODS: Twenty-seven patients were included in the study in which canine retraction was carried out with an elastomeric chain and NiTi closed coil spring on either side of the arch applying 150 g of force on both sides. GCF samples were collected before the commencement of canine retraction, on the first, seventh and 21st day after application of force and were analysed for leptin levels by the ELISA technique. Impressions for the study model were taken at baseline and after 21 days to measure the rate of tooth movement bilaterally with an electronic digital calliper. RESULTS: The results obtained within both groups showed a significant decrease in leptin levels from baseline to 21 days ( P = 0.0001). There was no significant difference in leptin levels between both groups, but leptin levels decreased more in the NiTi closed coil spring group. The rate of tooth movement is not significantly different between both groups, but space closure is faster in the NiTi closed coil spring group. CONCLUSIONS: Leptin levels in GCF showed a significant decrease during canine retraction using an elastomeric chain and NiTi coil spring; there was no significant difference in the rate of tooth movement between both the groups.

E-cadherin-mediated impairment increases anti-apoptotic mechanism through upregulation of Bcl-2: An immunohistochemical study in various patterns of invasion of oral squamous cell carcinoma.

BACKGROUND: Bcl-2 and E-cadherin proteins are known to be involved in the control of apoptotic cell death and invasive potential, respectively, which is an important hallmark of tumor regulation that influences their biologic behavior. AIM: This study investigates the relationship of Bcl-2 and E-cadherin immunoexpression in various Bryne's patterns of invasion. MATERIAL AND METHODS: Immunohistochemical analyses for Bcl-2 and E-cadherin were performed on paraffin-embedded tissue sections on 40 cases (32 cases of Oral squamous cell carcinoma and eight cases of controls) and were scored using qualitative and quantitative (percentage positive) analysis. STATISTICAL ANALYSIS: The resulting data were analyzed using SPSS software version 19. Correlation between patterns of invasion and qualitative scores of Bcl-2 and E-cadherin was calculated using Spearman rho correlation. Difference of mean percentage of positive cells of Bcl-2 and E-cadherin in different patterns of invasion was tested by ANOVA followed by Tukey HSD test. RESULTS: Bcl-2 and E-cadherin immunoreactivity was positively correlated with Bryne's pattern of invasion (P value<.05). An inverse relation was found between Bcl-2 and E-cadherin expression with Bryne's patterns 1-5 of invasion. CONCLUSIONS: The results pointed to the antagonistic role of E-cadherin and Bcl-2 and thus provide the opportunity for cell survival along with increased invasive potential.

Alpha-2-Macroglobulin Levels in Gingival Crevicular Fluid Pre- and Post-scaling and Root Planing with Adjunctive Tetracycline Fibers in Chronic Periodontitis: A Randomized Controlled Trial.

BACKGROUND: This split-mouth clinical study aimed to investigate levels of alpha-2-macroglobulin (a2M) in gingival crevicular fluid (GCF) of chronic periodontitis patients pre- and post-scaling and root planing (SRP) with or without adjunctive use of tetracycline fibers. MATERIALS AND METHODS: In 30 patients of chronic periodontitis, samples of GCF were collected from the gingival sulcus before SRP. Recording of clinical parameters was conducted. This was followed by local drug delivery (LDD) of tetracycline fibers in test sites. In control sites, no LDD was done. Second samples of GCF were taken 90 days after treatment. Samples of crevicular fluid were analyzed to determine the levels of a2m. RESULTS: A gain of clinical attachment (CAL) of 3.30 mm for SRP and LDD and for SRP alone was 1.62 mm (p < 0.001). The pocket probing depth was significantly decreased by 2.43 mm for SRP and LDD and for SRP alone was 1.61 mm (p < 0.001) after 90 days. Alpha-2-macroglobulin was significantly reduced in GCF by SRP and SRP and LDD after 90 days (p < 0.001). CONCLUSION: Clinical and biochemical variables showed a more favorable outcome when SRP was combined with LDD of tetracycline fibers in management of patients suffering from chronic periodontitis.

Histomorphometric and molecular characterization of stromal and mineralized components in fibro-osseous lesions.

BACKGROUND: Indistinct and analogous histopathological features of various fibro-osseous lesions make establishing a definitive diagnosis a challenge. There is a need for additional molecular and histochemical tools to support and differentiate these lesions in order to establish a concrete diagnosis. MATERIALS AND METHODS: A retrospective analysis of biopsied lesions in formalin-fixed paraffin-embedded sections (10 cases each of fibrous dysplasia, ossifying fibroma, and cement-osseous dysplasia) retrieved from the archives was studied for immunoexpression of osteocalcin (quantitative analysis in osteocytes), collagen characterization using Azan, Picrosirus, and Toluidine blue stain for evaluating intensity and localization of collagen fibers, and morphometric analysis of vasculature (for evaluating mean vessel density as square microns). RESULTS: Positive immunostaining of osteocalcin suggested mutations of the GNAS-1 gene found in fibrous dysplasia indirectly, as it is a negative regulator of bone formation. Osteocalcin immunopositivity was quantitatively measured in the fibro-osseous lesions, with fibrous dysplasia measuring 14.47 ± 3.628 as compared to ossifying fibroma measuring 5.23 ± 1.33, followed by cemento-osseous dysplasia measuring 2.30 ± 1.409. Toluidine blue suggests the presence of oxytalan fibers (resistant to acid hydrolysis) in ossifying fibroma and cemento-osseous dysplasia, pointing toward the pathogenesis of the lesion. Azan stain and Picrosirus (under a polarizing microscope) helped in distinguishing hard tissue characteristics (70% of cases of fibrous dysplasia showed only a magenta component followed by intermixed magenta with a blue component in 20% of cases and only 10% of cases showed magenta with blue borders whereas for ossifying fibroma, 40% of cases depicted magenta with blue borders along with the other 40% with intermixed magenta with blue component). The mean vessel density was also highest in fibrous dysplasia measuring 7.90 ± 1.079 (in Sq. micron area), followed by ossifying fibroma and cemento-osseous dysplasia. CONCLUSION: The diagnosis of fibro-osseous lesions by hematoxylin and eosin alone is confusing and thus should be supported by relatively simple histomorphometric analysis for better treatment outcomes. At the diagnostic stage of fibro-osseous lesions, evaluation of intralesional vessel size, reliable molecular marker, and histochemical nature can aid in differentiating fibrous dysplasia from central ossifying fibroma and cemento-osseous dysplasia alongside, other clinical, radiographic and pathological criteria. These parameters help in the diagnostic decision-making of fibro-osseous lesions.

Odontogenic myxoma of the mandible: An update on pathogenesis and differential diagnosis.

Odontogenic myxomas (OMs) represent asymptomatic, slowly expanding gnathic lesions with aggressive biological behaviour. Though the spectrum of OMs remains classical with multilocular radiolucency and presentation of stellate-shaped cells embedded in a mucoid stroma, they may mimic many other lesions radiographically or histopathologically. We hereby discuss a case of OM in a 28-year-old woman with special emphasis on pathogenesis and differential diagnosis.

Neuroectodermal influence in odontogenic cyst and tumor: evidence through CD56 immunoexpression.

BACKGROUND AND OBJECTIVES: CD56, associated with neuroectodermal differentiation of the embryonal cells, is often considered a marker of neural lineage. Odontogenic keratocysts (OKCs) are of particular interest because of their characteristic histopathologic features, high recurrence rate, and aggressive behavior. CD56 immunoreactivity in these lesions has been reported with very high frequency. The present study analyzes the immunoexpression of CD56 in ameloblastoma (AM) and OKC to infer neuroectodermal influence in the pathogenesis of odontogenic lesions and its correlation with clinicopathologic parameters. MATERIALS AND METHODS: Fifty histopathologically confirmed cases of OKC and AM, 25 from tooth-bearing (TB) and molar-ramus (MR) regions each, and 5 dental follicular tissues as control were collected from the department archives and immunohistochemical analysis with CD56 was carried out. RESULTS: CD56 immunopositivity was seen in 64% AM and 36% OKC cases. The majority of AM cases showed cytoplasmic expression in the peripheral cells of odontogenic islands; similarly, OKC cases showed continuous and uniform cytoplasmic expression in the basal and parabasal cells of the cystic lining. CD56 immunopositivity was found in more AM cases as compared to OKC cases in both the TB and MR regions. INTERPRETATION AND CONCLUSIONS: The assessment of CD56 immunoexpression in odontogenic cyst and tumor (AM) may aid in understanding the role of neuroectodermal influence in the etiopathogenetic pathways and a possible influence of CD56 on the clinical behavior and aggressiveness of the odontogenic lesions. A correlation of CD56 expression with the clinical outcome of the disease (site, perforation, root resorption, and tooth displacement) can help envisage possible prognostic assessment for these lesions.

Diagnostic utility of special stains in defining the spectrum of maxillofacial pathologies.

Background: The Fine needle aspiration cytology (FNAC) is considered as a valuable and distinguished diagnostic test in the initial assessment of the patients presenting with a mass in the head and neck region or when a recurrence is suspected after previous treatment. Aims: This study was therefore designed to elucidate the efficacy of FNAC as an alternate diagnostic tool to histopathology in head and neck swellings and evaluation of staining efficacy of PAP and MGG stain over Haematoxylin and eosin (H and E) in routine cytopathological smears. Settings and Design: The study was conducted in the Department of Oral and Maxillofacial Pathology, where FNAC samples were collected from 150 patients with head and neck swellings. Materials and Methods: All the slides were stained with H and E, Papanicolaou (PAP), and May Grunewald Giemsa (MGG) stains. The cytopathological diagnosis was compared with histopathological diagnosis based on H and E stained sections obtained from paraffin-embedded formalin-fixed biopsy specimen of benign and malignant neoplasms. Statistical Analysis Used: The resulting data were analyzed using SPSS software version 19. Differences between the variables were analyzed using Pearson Chi-square test and Kruskal-Wallis test wherever applicable. Results: The FNAC as a diagnostic tool has sensitivity of 84.8%, 72.72%, and 78.78%, specificity of 62.5%, 75%, and 75%, and accuracy of 80.48%, 73.14%, and 78.04% in H and E, MGG, and PAP stain, respectively. PAP stain was the most efficient stain when all qualitative parameters are taken into consideration with maximum sensitivity and specificity for achieving definitive cytodiagnosis. Conclusions: The FNAC is an inexpensive and minimally invasive technique to diagnose different types of head and neck swellings and complement histopathological diagnosis.

Emerging insights of NK cells immunosurveillance in histomorphologic prognostic indicators of oral squamous cell carcinoma.

Background: IFN-gamma and natural killer (NK) cells have been considered the most effective cells in the combat of cancer, contributing to better prognosis and longer survival. The aim of the study was to analyze and correlate the CD 57 immunopositive NK cell-mediated Interferon-γ pathway in regulating immune mechanisms in Oral Squamous Cell Carcinoma. Materials and Methodology: The study sample was composed of a total of 40 cases of histopathologically confirmed cases of Oral Squamous cell carcinoma (OSCC). Clinical data such as age, gender, habit history, signs and symptoms, and TNM staging were obtained for each case. The biopsy specimens of the cases obtained were fixed with 10% neutral buffered formalin and processed and embedded in paraffin wax. 3-4 µ thick sections were taken for hematoxylin and eosin staining and immunohistochemistry procedure. A saliva sample was collected from each patient and stored at 20 degree Celsius for estimation of salivary interferon-gamma levels using the sandwich ELISA technique. Results: CD 57 NK cells quantitative assessment was significantly associated with tumor budding, cell nest size, the pattern of invasion, lymphocytic host response, NK cell morphology, Depth of invasion, and Tumor thickness. The ratio of CD 57 immunopositive NK cells to salivary IFN-γ levels showed a significant association with histopathological grades, tumor size, and lymph node status. Conclusion: Adoptive cellular transfer therapy with NK cells has been advocated in both experimental models and clinical trials in treating hematopoietic malignancies. The strategy is based on reviving the patient innate immune surveillance and control of tumor invasion by the infusion of activated NK cells. The IFN-gamma and NK cell infiltration in oral squamous cell carcinoma might show a distinctive tumor microenvironment with a favorable local cytotoxic immune response against neoplastic cells.

Potentiated action on the progression of OSMF by hypoxia mediated signaling pathway by the epithelial mesenchymal transition and angiogenic apparatus.

Background: Epithelial-mesenchymal transition (EMT) is a complex process, in which epithelial cells acquire the characteristics of invasive mesenchymal cells. EMT has been implicated in cancer progression and metastasis as well as the formation of many tissues and organs during development. Aim: The aim of the study was to ascertain the role of hypoxia-mediated signaling pathways influencing EMT and angiogenesis in progression of oral submucous fibrosis (OSMF). Materials and Methods: Evaluation of the immunoexpression of alpha-smooth muscle actin (α-SMA), E-cadherin, vimentin, and factor VIII receptor antigen in OSMF and oral squamous cell carcinoma (OSCC) arising from OSMF was done. Differences between the different variables were analyzed using ANOVA test and Pearson's Chi-square test, and Mann-Whitney test was also calculated. Results: The mean α-SMA positive myofibroblasts increased from Group 1 (OSMF) to Group 2 (OSCC), especially those in the deeper connective tissue stroma. The mean labeling index of vimentin and mean vessel density immunoexpression was more in Group 2 (OSCC) as compared to Group 1 (OSMF). Mean α-SMA correlated negatively with E-cadherin expression and positively with vimentin and factor VIII immunoexpression. E-cadherin expression correlated negatively with factor VIII and positively with Vimentin expression. Conclusions: The molecular mechanisms responsible for the development of OSCC in patients with OSMF require unification of multiple progressive pathogenetic mechanisms involved in the progression of the disease.

Pathogenetic model of survivin-dependent molecular signalling pathways in tumorigenesis of oral cancer and precursor lesions.

Background: p53 tumour suppressor gene limits unchecked cellular growth in response to DNA damage, by causing G1 arrest and the activation of apoptosis. Inhibitors of apoptosis include survivin which acts by inhibition of caspases. Survivin has a significant role as a cell cycle modulator and is only minimally present in mature tissues. Aberrant expression of p53 and survivin has been evaluated in various carcinomas. Thus, the objective of this research was to elucidate the co-expression of p53 and survivin in tissue samples of Oral Potentially Malignant Disorders (OPMDs) and Oral Squamous Cell Carcinoma (OSCCs). Method: Thirty tissue samples of OPMDs and 30 tissue samples of OSCCs taken from department archives were used in the study. Expression of p53 and survivin was analyzed in the study groups by the help of immunohistochemistry. Also, co-expression of both the markers was evaluated. Results: The expression of p53 and survivin in the oral epithelium of patients with OSCCs was significantly higher than that in patients with OPMDs (P value ≤0.05). Conclusion: Our results provide insights into the altered survivin and p53 co-expression with significant immunoexpression within the study groups. Therefore, survivin and p53 could be better markers for identifying cell proliferation and apoptotic pathway. Also, malignant transformation rate of OPMD increases with increased expression of these markers.

Immunolocalization of epithelial cell adhesion molecule and matrix metalloproteinase-9 in oral epithelial dysplasia and oral squamous cell carcinoma.

INTRODUCTION: Cancers are complex tissues composed of multiple distinct cell types that participate in heterotypic interactions with one another. Physiologically cell-to-cell contacts formed by dense populations of normal cells operate to suppress further cell proliferation. OBJECTIVES: The objective of the study is to evaluate and compare the immunoexpression of matrix metalloproteinase-9 (MMP-9) and epithelial cell adhesion molecule (EpCAM) in oral epithelial dysplasia (OED) and oral squamous cell carcinoma (OSCC) and to hypothesize their role in the progression in varying grades of these lesions. MATERIALS AND METHODS: A total of 60 samples comprising of 30 cases each of OED and OSCC. Three micrometers thin sections were taken and subjected for hematoxylin and eosin stain and immunohistochemical procedure. The sections were incubated with monoclonal anti-EpCAM anti-MMP-9 antibody. The data were analyzed using SPSS software version 19. RESULTS: The results of the study show EpCAM immunoexpression decreased in OSCC when compared to OED. MMP-9 immunoexpression increased in OSCC when compared to OED (statistically significant, P ≤ 0.05). CONCLUSION: Correlation between EpCAM and MMP-9 may help to unravel the signaling cascades involved in the carcinomatous changes, tumor cell invasion, and progression of OSCCs.

Embryological Basis of Cystic Hygroma: A Case Report.

Aim: We intend to present a cystic hygroma (CH) case in a 2-year-old baby in the anterior cervical triangle, which is a rare site, as the most common site for the occurrence of CH is the supraclavicular fossa of the posterior cervical triangle. Background: Among developmental abnormalities in the lymphoid system, CH are usually seen in the posterior neck. Lymphatic malformations are generally exhibited either at birth or before the age of 2 years. Lymphatic channels are attenuated endothelium-lined spaces devoid of any cells and smooth muscle layer. Also, morphologically distinguishing normal lymphatic channels from venules or capillaries is a challenge. Case description: A 2-year-old female patient reported having a chief complaint of swelling in the left submandibular region for 4 days. The patient underwent surgery for CH 18 days after birth. Swelling was rubbery in texture and firm in consistency. Conclusion: A D2-40 immunoexpression was an identifying clue for normal lymphatics in comparison to morphology. Henceforth, this can be concluded that such tumors depict at least partial differentiation of endothelial cells lining lymphatic spaces. Clinical significance: The present article helps in illuminating the role of D2-40 in the diagnosis of lymphatic malformations, such as CH, and also highlights the embryological basis of the pathogenetic mechanism of this rare disease, which potentiates the role of various treatment modalities in pediatric cases for management considerations. How to cite this article: Yadav S, Gulati N, Shetty DC, et al. Embryological Basis of Cystic Hygroma: A Case Report. Int J Clin Pediatr Dent 2022;15(6):774-778.

Histopathological prognosticators and their clinicopathological correlation in oral squamous cell carcinomas of the tongue.

Background: Tongue carcinomas account for 25%-40% of intraoral squamous cell carcinomas (OSCCs). Although TNM staging systems is an international standard for cancer reporting, prognosis evaluation, and treatment planning, multiple histopathological risk assessment predictors such as tumor thickness (TT), tumor shape, tumor growth pattern, and invasive malignancy grading scoring systems have been studied and should form a basis for prediction and prognostication of such aggressive carcinomas. Aim: To evaluate and characterize the histomorphological prognostic indicators in OSCCs of tongue and compare it with OSCCs of other anatomic sites within the oral cavity. Furthermore, to elucidate the significance of histopathological indicators in predicting prognosis of tongue squamous cell carcinomas (SCCs). Materials and Methods: Forty SCC cases with 20 each of tongue and 20 from other intraoral sites were retrieved from department archives. Clinical data and staging were obtained for each case. Histomorphological parameters including pattern of invasion (POI), tumor budding (TB), depth of invasion (DOI), TT, lymphocytic host response, tumor-associated tissue eosinophilia (TATE), vascular invasion, perineural invasion (PNI), and muscular invasion were assessed. The results were statistically evaluated. Results: TB, DOI, and sarcolemmal spread were significant histologic predictors in tongue SCC. Upon correlation of histomorphological parameters with clinical staging, TT, POI, and TATE were observed to be significantly correlated (P ≤ 0.05). Conclusion: The histomorphological risk assessment model may serve as important addition to the existing prognosticators and may be used as a prognostic index to help plan and individualize treatment protocol in cases with aggressive high-risk disease for whom the use of multimodality treatment seems beneficial.

HIF - 1α as a marker of risk assessment for malignant transformation in oral submucous fibrosis: An immunohistochemical study.

Background: Oral submucous fibrosis (OSMF) is a potentially malignant disorder associated with habit of chewing betel quid containing arecanut. Morphological features of OSMF especially fibrosis suggests a possibility of the hypoxic environment in diseased tissues. The adaptation of cells to hypoxia appears to be mediated via hypoxia inducible factor-1α (HIF-1α) which is also said to be associated with malignant transformation of epithelial cells in various other carcinomas like prostate and cervical carcinoma. Therefore, the main objective of this study was to investigate the role of HIF-1α in progression and malignant transformation of OSMF. Materials and Methods: The study group consisted of histo-pathologically diagnosed 30 cases of oral submucous fibrosis and 10 cases of OSCC were taken as control. The immunohistochemistry was carried out on neutral buffered formalin-fixed paraffin-embedded tissue sections by using the monoclonal antibody of HIF-1α. Statistical analysis was done using SPSS software version 2.0. Results: A gradual and significant rise in the expression of HIF-1α was observed in various grades of OSMF and OSCC cases. HIF 1α expression was increased in cases showing hylanization and constricted blood vessels. A cut off value of 39.6% of HIF-1α positive cells was determined statistically to categorize the cases into high risk and low risk group for malignant transformation. Conclusion: Overexpression of HIF-1α may contribute to the progression and malignant transformation of OSMF. Cases expressing more than 40% of HIF-1α positive cells are at a greater risk for malignant transformation.

MCM3 proliferative index is worthier over Ki-67 in the characterization of salivary gland tumors.

BACKGROUND: Salivary gland tumors bear uncanny characteristics of being different based on their morphological aspects rather than the presence of clear demarcation. This ambiguity in the spectrum from benign to malignant salivary gland neoplasms while categorizing the neoplasm is having inherent pitfalls. The present study was, therefore, designed to characterize benign and malignant salivary gland tumors based on their proliferative indices. MATERIALS AND METHOD: Study samples comprised of 97 cases of histopathologically confirmed benign and malignant salivary gland tumors. The cases were immunohistochemically assessed for MCM3 and Ki-67 expressions and the molecular characterization was performed based on the findings. RESULTS: The majority of benign and malignant salivary gland tumors were from the parotid gland, (51.2%) and (42.4%), respectively. Overall mean labeling index of MCM3 was higher i.e., (5.60 ± 3.99) in comparison to Ki-67 i.e., (2.82 ± 3.14) with P = 0.05 using paired t-test. Besides, malignant salivary gland neoplasms represented a higher mean score of MCM3 and Ki-67 than benign neoplasms. CONCLUSION: The requirement of a novel marker has led to the use of MCM3 which has a characteristic role in the entire spectrum of the cell cycle. The present study highlighted the extrapolation of MCM3 over Ki-67 for diagnosis and for true characterization of biologic behavior of salivary gland pathologies which may, in turn, influence the treatment modality employed for such lesions.

"Neuroectodermal influence of CD 99 immunoexpression correlates with the clinical behavior of odontogenic cysts and tumors".

Background: Odontogenic tumors show a variety of characteristic features that are dependent on the tumor cell origin and the stage of tumor cell differentiation. Odontogenic cysts arise from the enamel organ or remnants of dental epithelium which influences their pathogenetic model and further clinical behavior of these lesions. Aim: The study aims at assessment of CD 99 immunoexpression in odontogenic keratocyst (OKC) and ameloblastoma (in tooth bearing [anterior to third molar] and nontooth bearing areas [molar ramus area]) to postulate neural influence in their pathogenesis and the clinical behavior. Materials and Methods: Immunohistochemical analysis for CD 99 was performed on paraffin-embedded tissue sections on 50 histopathologically confirmed cases of OKC and ameloblastoma (25 each) arising within the oral cavity and were scored qualitatively, topographically, and according to cellular localization. Statistical Analysis: The resulting data were analyzed using the SPSS software version 20.0. The significance of the parameters was tested by the Pearson's Chi-square test (P ≤ 0.05 as statistically significant). Results: CD99 immunoreactivity was distributed in both tooth bearing and nontooth bearing groups of OKC and ameloblastoma with an increased immunoexpression in basal and suprabasal layers of OKC in nontooth bearing area and in peripheral cells of ameloblastoma in nontooth bearing area confined to the cell membrane. Conclusions: The results point toward the role of CD99 in the pathogenesis and aggressive behavior of such odontogenic lesions and it can be used as a promising therapeutic target.

Tumor-associated tissue eosinophilia versus tumor associated blood eosinophilia: A ratio of diagnostic importance in oral squamous cell carcinoma.

INTRODUCTION: Eosinophils are multifunctional granulocytes, which play a pivotal role in health and disease. Tumor Associated Tissue Eosinophilia (TATE) has long been evaluated in the diagnosis and progression of oral squamous cell carcinomas (OSCCs). However, their association with Tumor Associated Blood Eosinophilia (TABE) in OSCCs is still far fetched. We, therefore, attempted to evaluate their individual roles and to achieve a ratio between TATE and TABE in order to signify its usage in objectifying the diagnosis. MATERIALS AND METHODS: TATE was evaluated using H and E stain per 10 high power fields in 33 previously diagnosed cases of OSCC which were retrieved from department archives. TABE values were achieved from complete blood hemogram reports of patients. TATE/TABE ratio was calculated. All the parameters were clinicopathologically correlated and statistically evaluated using SPSS. RESULTS: TATE represented higher values in well-differentiated squamous cell carcinoma (WDSCC) and poorly differentiated squamous cell carcinoma (PDSCC) and was least in moderately differentiated squamous cell carcinoma (MDSCC), whereas TABE linearly increased from WDSCC to PDSCC. TNM Stage II cases revealed the highest TATE and lowest TABE. TATE/TABE ratio was the highest in WDSCC. CONCLUSION: Due to the dual nature of eosinophils in early and late carcinogenesis events, evaluation of only TATE might not be conclusive in determining tumor grade. Hence, in a first of its kind attempt, the TATE/TABE ratio may be suitable to achieve a criterion for the determination of tumor grade and may also help to unfold the underlying biologic events.

Staining efficacy assessment of a differential routine and special stains for pathological stromal calcifications in maxillofacial lesions.

Head and neck connective tissue lesions may have diverse calcifications within the fibrous connective tissue stroma. The perplexity involved in the identification and determination of the nature or degree of calcification through routine hematoxylin and eosin (H&E) stains necessitates the usage of a specific, simple, and cost- and time-effective differential staining techniques. The aim of the present study was to develop criteria to distinguish bone formation from bone resorption using methylene blue-acid fuchsin (MB/AF) stain and the role of collagen fibers in the identification of stromal calcifications using polarizing microscopy with picrosirius red stain. Twenty cases with pathological diagnoses for various stromal calcifications in maxillofacial lesions were retrieved from the departmental archives. Decalcified formalin fixed paraffin embedded tissue sections were stained with hematoxylin and eosin, Masson's trichrome (MT), methylene blue-acid fuchsin (MB/AF), and picrosirius red. The stained sections were assessed to identify the calcifications found in the surrounding connective tissue stroma. It was observed that most cases showed maximum staining intensity with MB/AF stain as compared to the other staining methods. Moreover, the results suggested that contrast between calcification and stromal soft tissue was best distinguished with the MB/AF stain except in the case of dystrophic calcifications. Along with this, polarizing microscopy with picrosirius red enables better characterization of stromal components. Although the H&E stain and a connective tissue stain i.e. Masson's trichrome, are employed routinely in histopathology; the use of special stains such as MB-AF and picrosirius red facilitates the identification of calcifications from the stromal tissues.

Molecular Analysis as a Guide to Determining the Extent and Pathophysiology of Perilesional Tissues in Oral Epithelial Dysplasias.

INTRODUCTION: Clinically and histologically normal appearing perilesional mucosa of epithelial dysplasia may harbor early genetic changes. Hence, the present study is designed to determine the early molecular changes in the form of p16 and EGFR immunoexpressions in perilesional tissues. OBJECTIVES: To analyze immunohistochemical expressions of p16 and EGFR individually and percentage change of immunoexpressions in oral dysplastic lesions and their perilesional tissues. MATERIALS AND METHODS: Forty formalin-fixed paraffin-embedded tissues of oral epithelial dysplasia with perilesional tissue marked by India ink were included in this study. Immunohistochemical staining was performed using anti-p16 and anti-EGFR monoclonal antibodies (BioGenex) using squamous cell carcinoma of uterine cervix and breast carcinoma as the positive controls, respectively. RESULTS: p16 and EGFR expressions were assessed based on the presence, intensity, extent and immunolocalization of positive cells. Out of 40 cases, p16 immunoexpression was positive in 82.5% cases of lesional tissues and in 62.5% cases of perilesional tissues (p ≤ 0.05); however, EGFR immunoexpression was positive in 90% cases of both lesional and perilesional tissues (p > 0.05). CONCLUSION: The disease status and progression based on p16 and EGFR expressions and co-expressions can be used as an effective guide to evaluating the progression of normal epithelium to dysplastic epithelium in otherwise clinically normal mucosa.

Immunohistochemical expression of p53 and murine double minute 2 protein in odontogenic keratocyst versus variants of ameloblastoma.

INTRODUCTION: Oncogenes and tumor suppressor genes play a major role in cancer formation, growth, and progression. One of the important findings in this area is that murine double minute 2 (MDM2) oncogene is a negative regulator of wild-type p53. In tumors, expressing wild-type p53, inhibition of MDM2 expression will stabilize p53 and allow it to perform its proapoptotic function, while simultaneously preventing MDM2 from exerting its p53-independent oncogenic effects. The intracellular levels of p53 are tightly regulated by MDM2, as it is a key player in autoregulatory feedback loop under nonstressed conditions. The p53-MDM2 relationship is vital not only for essential functions of the cell, but it also appears to be an integrated part of the complex cellular network which supports the importance of this affair and is a hallmark for its coexistence. SUBJECTS AND METHODS: This study was designed to identify immunohistochemically the expression of p53 and MDM2 gene using monoclonal antibody in 60 cases of formalin-fixed paraffin-embedded tissue blocks, of which 20 cases were of solid multicystic ameloblastoma (SMA), 20 cases were of odontogenic keratocyst (OKC), and 20 cases were of unicystic ameloblastoma (UA). RESULTS: Immunoexpression of p53 and MDM2 was highest in OKC followed by SMA and was minimum in UA. Further results showed positive correlation between both the molecules. CONCLUSION: The studied showed that the relationship has a significant role in cancer etiology and progression and therefore is an important topic for future research which should help in the development of new therapeutic agent against cancer.