Expression of CD4+ and CD8+ Tumor-Infiltrating Lymphocytes in Oral Squamous Cell Carcinoma and Their Relationship With Clinicopathological Parameters: A Cross-Sectional Study.

Background Oral squamous cell carcinoma (OSCC) is the most common malignant neoplasm of the oral cavity. The tumor microenvironment (TME) is a dynamic ecosystem composed of components contributed by both the tumor and the host. The immune cells of TME, mainly CD4+ and CD8+ tumor-infiltrating lymphocytes (TILs), suppress the proliferation of cancer cells and play a crucial role in the progression of OSCC. The present study aims to analyze the immunohistochemical expression of CD4+ and CD8+ TILs in OSCC and to compare and correlate them with clinicopathological parameters. Methodology A total of 75 formalin-fixed paraffin-embedded samples of cases diagnosed with primary OSCC were immunostained with CD4+ and CD8+ antibodies and their expression was compared with the clinicopathological parameters. Results There was a significant positive correlation between CD4+ and CD8+ expression (r = 0.655, p = 0.001). Both CD4+ (r = -2.37, p = 0.041) and CD8+ (r = -0.348, p = 0.002) expressions negatively correlated with the TNM stage (r = -2.37, p = 0.041) of OSCC. CD8+ expression positively correlated with histopathological grade (r = 0.288, p = 0.012). Conclusions The study findings suggest that CD4+ cells are essential to maintain and sustain CD8+ TIL-mediated anti-tumor response. CD4+ and CD8+ TILs are key players in cell-mediated adaptive immunity and prevent tumor progression and metastasis. Strikingly, the higher grade of tumors despite heavy CD8+ infiltration may possibly be due to cancer immunoediting.

Age Estimation using Cephalometrics-A Cross-Sectional Study among Teenagers of Salem District, Tamil Nadu.

Background: Age estimation is important not only in identifying dead body of a person but also in living persons since there is an increasing rate of juvenile delinquencies recorded every year. To avoid foul play by age fabrication, legal age estimation becomes important. Facial growth alteration takes place in the jawbones as age advances which can be observed with lateral cephalometry. Thus, the aim of the study is to create a regression formula for age estimation using cephalometrics of teenagers in Salem population. Materials and Methods: A cross-sectional study was done using 770 lateral cephalometrics of teenagers (13-19 yrs) in Salem population. Nine cephalometric points with two linear hard tissue measurements (condylion to mandibular plane (AFH) and palatal plane to menton (PFH)) and one angular soft tissue measurement (z angle) were recorded as predictor variables using a digital lateral cephalometric software (Carestream CS8100 SC) which were subjected to regression analysis using SPSS version 21.0 to develop a formula for age estimation. Results: Significant association on age was obtained for the two linear measurements. The regression formula generated for estimating the age was Age = 7.146 + 0.044 (AFH) + 0.146 (PFH) with R2 value = 0.674. Conclusion: Within the limitations of the present study, age estimation of teenagers in Salem population can be estimated. The predictability of the age can be increased by taking more cephalometric variables in generating the formula with increase in sample size.

Immunohistochemical expression of putative cancer stem cell markers aldehyde dehydrogenase 1, SOX2, CD44 and OCT4 in different grades of oral epithelial dysplasia.

Objectives: The hypothesized existence of cancer stem cells (CSC) and its markers aldehyde dehydrogenase 1 (ALDH1), CD44, SOX2 and OCT4 in oral dysplastic tissues provides the potential for a more reliable assessment of malignant transformation of oral epithelial dysplasia (OED). Thus, the present study is intended to evaluate the immunohistochemical expression of four different CSC markers ALDH1, CD44, SOX2 and OCT4 in different grades of OED and to investigate the co-expression of these putative stem cell markers in OED. Subjects and Methods: A total of 35 samples of varying grades of OED which included 7 mild, 11 moderate and 17 severe dysplasia samples and 10 samples of normal oral mucosa without dysplasia were used. Four sections each from all 45 samples were stained with ALDH1, CD44, SOX2 and OCT4, respectively, by immunohistochemistry. The acquired data were analyzed and evaluated using the Chi-square test and unpaired t-test and the P < 0.05 was taken significant. Results: ALDH1 and SOX2 expression percentages showed statistically significant differences among study groups (P < 0.05). Statistical comparison of percentage expression of CD44 and OCT4 between OED and normal was nonsignificant (P > 0.05). Co-expression of all four markers was found in 15 cases of OED with none of the normal cases showing co-expression. Conclusion: The expression of CSC markers in OED and normal mucosa differs significantly with co-expression of all four markers located only in dysplastic tissues. Until now, no single protein marker has been able to unequivocally identify the CSCs. Thus, a panel of putative CSC markers will help in identifying the patients with high risk for malignant transformation in OED.

Evaluation of aryl hydrocarbon receptor expression in oral squamous cell carcinoma and normal oral mucosa using western blot.

BACKGROUND: Aryl hydrocarbon receptor (AHR) is a ligand-activated transcription factor that acts as a binding site for toxic chemicals, particularly the dioxin group of chemicals. Elevated levels of AHR have been observed in various human cancers, including lung carcinomas, hepatic carcinomas and in mammary tumors. However, the expression of AHR in oral squamous cell carcinoma (OSCC) patients who are tobacco users are less explored. AIMS AND OBJECTIVES: The aim of the present study is to evaluate and compare AHR levels in OSSC patients and in normals using Western blot technique in an attempt to explore the possible role of AHR in oral carcinogenesis. MATERIALS AND METHODS: The study sample consisted of ten oral squamous cell carcinoma cases which were diagnosed clinically and confirmed histopathologically as OSCC and four samples of the normal oral mucosa. AHR protein expression was evaluated using Western blot technique and chemiluminescence detection kit. The densitometry was performed on a Microtek scan maker MSP flatbed scanner and quantified using Image J software. Mean AHR protein levels were calculated and compared between OSCC and normal oral mucosa using Student's t-test. RESULTS: The mean AHR protein level in OSCC samples (n = 10) was 2878.90 ± 1231.27 and 975.75 ± 227.27 in the normal oral mucosa (n = 4). The OSCC samples showed significantly higher levels of AHR protein compared to the normal oral mucosa (P = 0.008). CONCLUSION: The study showed a significantly higher expression of AHR in oral squamous cell carcinoma samples when compared to the normal oral mucosa, suggesting a possible role of AHR in the initiation, promotion and progression of oral squamous cell carcinoma.

Sex determination using maxillary arch width of pediatric population of Namakkal district, India: A forensic study.

AIM AND OBJECTIVES: The aim of the study is to compute a new formula for sex determination using maxillary arch width of a pediatric population of Namakkal district. MATERIALS AND METHODS: The sample consisted of 146 females and 218 males of South Indian origin aged between 4 and 6 years. Alginate impressions of the upper and lower dental arch were made, and casts were poured immediately. A digital vernier caliper was used to obtain measurements. Arch width at canine, first molar and second molar for both maxilla and mandible were considered for measurement. Statistical analysis was performed using the Statistical Package for the Social Sciences Version 20.0 software. RESULTS: The Student's t-test was used to find out the significance between male and female among the different predictor variables at P < 0.05. Significant sexual dimorphism was found in maxillary intercanine width and maxillary first and second intermolar width with conical discriminant function coefficient of 0.732, -0.177 and -0.244, respectively. CONCLUSION: The formula derived from the present study could be of great value in sex determination of pediatric populations of Namakkal district.

Predicting Tumor Sensitivity to Chemotherapeutic Drugs in Oral Squamous Cell Carcinoma Patients.

Oral Squamous Cell Carcinoma (OSCC) patients respond poorly to chemotherapy. We analyzed the expression of 11 drug response-related genes in 31 OSCC biopsies, collected prior to any treatment, using custom-designed PCR array. Further, we investigated the drug response pattern of selected anticancer drugs by BH3 (Bcl2 Homology-3) profiling in the primary cells isolated from OSCC tissues. Then, we correlated the results of drug-response gene expression pattern with apoptotic priming to predict tumor response to chemotherapy. The best performing drug (BPD) and response differences (RD) between the drugs were identified using statistical methods to select the best choice of drug in a personalized manner. Based on the correlation, we classified OSCC tumors as sensitive (13 tumors), moderately responsive (16 tumors) or resistant (2 tumors) to chemotherapy. We found that up-regulation of genes linked with drug resistance facilitates survival of tumor samples, which was revealed by the percentage of apoptotic priming. Moreover, we found that paclitaxel-induced 40-45% apoptotic priming compared to other drugs. Average response difference (RD) analysis showed that 80% of tumors responded well to paclitaxel as compared to other drugs studied. Therefore, gene expression analysis with BH3 profiling reveals drug sensitivity that could be translated for drug selection before treatment.

Buccal Micronuclei Assay as a Tool For Biomonitoring DNA Damage in Oral Lichen Planus.

INTRODUCTION: The malignant transformation rate of Oral Lichen Planus (OLP) is between 0% and 5.8%. Oral lesions of lichen planus clinically presents itself multifocally, simulating the process of field cancerization in high risk malignancies. The Buccal MicroNucleus Cytome Assay (BMN Assay) provides a platform to identify the high risk individuals by evaluating the markers of nuclear damage at an earliest micro invasive phase. AIM: To evaluate DNA damage in exfoliated buccal mucosal cells in individuals with oral lichen planus lesions and thereby to delineate the high risk group. MATERIALS AND METHODS: Buccal smears from 22 OLP and 10 control samples were stained in modified Feulgen-Rossenback reaction for micronuclei assay. Cytological evaluation of number of MicroNucleated cells (CMN), Total Number of Micronuclei (TMN) in micronucleated cells was done in both groups. RESULTS: Frequency of micronucleated cells (CMN) when compared among the study and control group, a mean value of 4.27 ± 1.80 and 0.90 ± 0.88 were obtained respectively. On comparing the total number of micronuclei in the micronucleated cells (TMN) between the study and control groups, a mean value of 5.38 ± 2.42 and 1.5 ± 0.88 were obtained respectively. CONCLUSION: There was a significant increase in the frequency of micronuclei and the micronucleated cells in the oral lichen planus as compared to normal individuals.

Stem cells: An insight into the therapeutic aspects from medical and dental perspectives.

The recent advancements in the field of stem cell (SC) biology have increased the hope of achieving the definitive treatments for the diseases which are now considered incurable such as diabetes, Parkinson's disease and other chronic long standing conditions. To achieve this possibility, it is necessary to understand the basic concepts of SC biology to utilize in various advanced techniques of regenerative medicine including tissue engineering and gene therapy. This article highlights the types of SCs available and their therapeutic capacity in regenerative medical and dental fields.

Evaluation of myofibroblasts in oral squamous cell carcinoma using H1 calponin: An immunohistochemical study.

BACKGROUND: Oral squamous cell carcinoma (OSCC) is the most common malignancy of the oral mucosa. Stromal myofibroblasts play an important role in tumor invasion and metastasis, due to its ability to modify the extracellular matrix. The purpose of this study was to evaluate and compare the presence of myofibroblasts in normal mucosa, early invasive carcinoma and different grades of OSCC. MATERIALS AND METHODS: The study included the archival tissues of 18 OSCC of well, moderate and poorly differentiated grades, three early invasive carcinomas and five normal mucosa. Myofibroblasts were identified by immunohistochemical detection of h1 calponin. RESULTS: The percentage and intensity of h1 calponin were examined and positive immunostaining was observed in the myofibroblasts of all SCCs and early invasive carcinomas; however, these cells did not stain in the normal epithelium specimens. The presence of myofibroblasts was significantly higher in invasive pattern of OSCCs compared to normal mucosa cases (P < 0.070). A significant difference was not observed between the different grades of OSCC (P ≤ 0.812). CONCLUSION: These findings show the presence of myofibroblasts in OSCC but not in normal mucosa, suggesting that the genetically altered epithelium (carcinomatous epithelium) may have an inductive effect on the adjacent stroma to produce myofibroblasts. Also transdifferentiation of myofibroblasts is induced somewhere in the invasive stage of SCC irrespective of the epithelial cell differentiation.

Myoepithelial Cells (MEC) of the Salivary Glands in Health and Tumours.

Myoepithelial cells (MEC) are found in the secretory units of many mammalian exocrine glands such as mammary, sweat, lacrimal and salivary glands. They are interposed between the secretory cells and the basal lamina. Immunohistochemically they are found to contain keratin intermediate filaments and are, therefore, considered to have an epithelial origin but at the same time they contain a large number of myofilaments which represent a massive expression of contractile proteins such as actin, myosin, calponin and caldesmon. Thus have smooth muscle like property also and hence the name. Numerous functions of MEC have been described, the most important of them being important for contraction of the glands and recently it has been found to prevent tumour progression. It should be noted that the diversity in the occurrence and dilemma regarding the pathogenesis of salivary gland tumours is due to lack in uniformity regarding the cells participating in its oncogenesis, especially the MEC. Also proper and extensive studies regarding MEC are very limited and thus have posed difficulty for a pathologist to understand this cell. In this review we try to bring about a thorough description of this cell in both physiological and pathological aspects.

Evaluation of ploidy status using DNA-image cytometry of exfoliated mucosal cells in oral lichen planus.

BACKGROUND: Oral lichen planus (OLP) is one of the potentially malignant disorders (PMDs) with a malignancy rate of 0.2-2%. Aneuploidy is considered to be one of the important markers for malignant transformation and DNA-image cytometry (DIC) has been successfully employed in oral mucosal PMDs and also in tumors of the cervix, lung and biliary tract. AIMS: In this study, we intend to assess the ploidy status of exfoliated cells in OLP using DIC. MATERIALS AND METHODS: Exfoliated cells from 48 patients with different subtypes of OLP (reticular, plaque type, erosive and atrophic) and 10 controls were stained using Feulgen reaction and assessed for integrated optical density using image analysis software and the ploidy status was assessed. RESULTS: All the patients in the control group and most of the patients (93.5%) who had reticular or plaque type of OLP (29 out of 31) exhibited diploid nuclei in the smears, whereas 11 patients who had erosive or atrophic types of OLP showed aneuploid nuclei. CONCLUSIONS: The patients with erosive or atrophic types of OLP are at more risk and assessment of ploidy status by exfoliative cytology can be used as an adjuvant for diagnosis.

Cytomorphometric analysis of exfoliated cells in oral lichen planus.

BACKGROUND: Tumors are distinguished from normal tissues partly by their pronounced variability of cellular and nuclear dimensions. Therefore, such factors may be an indicator to assess whether the cells are malignant or not. Exfoliative cytology is a reliable tool in assessing such changes in the uterine cervix and has been used in the oral cavity also with success. The aims and objectives of the following study were to evaluate the malignant changes by assessing the quantitative parameters such as cytoplasmic diameter, cytoplasmic perimeter and cytoplasmic area (CD, CP, CA) and nuclear diameter, nuclear perimeter and nuclear area (ND, NP, NA) and cytoplasmic to nuclear ratio in the exfoliated cells of various subtypes of oral lichen planus (OLP) using cytomorphometry. MATERIALS AND METHODS: Oral exfoliated cells from nineteen cases of histologically proven OLP (1 atrophic, 13 reticular, 4 erosive and 1 plaque) and ten controls with healthy mucosa were taken and stained by Feulgen-Rossenback reaction and cytomorphometric analysis was performed using an image analysis software. The parameters taken into account were CD, CP, CA and ND, NP, NA. Furthermore CA/NA was calculated. The parameters were statistically analyzed using the t-test. RESULTS: Cytomorphometric analysis of all the parameters showed no significant difference between the control group and the reticular/plaque subtypes, whereas statistically significant (P < 0.05) differences was obtained between the control group and the atrophic/erosive subtypes group when compared using t-test. CONCLUSIONS: The cytomorphometric analysis of OLP shows that erosive/atrophic subtypes of OLP are at more risk and exfoliative cytology and cytomorphometry can be used as a tool to assess the malignant changes.

Screening of chemopreventive effect of naringenin-loaded nanoparticles in DMBA-induced hamster buccal pouch carcinogenesis by FT-IR spectroscopy.

The aim of the present study is to investigate the chemopreventive effects of the prepared naringenin-loaded nanoparticles (NARNPs) relative to efficacy of free naringenin (NAR) in modifying the functional, structural, and compositional changes at the molecular level during 7, 12-dimethylbenz[a]anthracene (DMBA)-induced hamster buccal pouch (HBP) carcinogenesis by Fourier transform infrared (FT-IR) spectroscopy. The results revealed that a significant increase in the amount of proteins and nucleic acid contents and a decrease in the amount of lipids and glycogen contents are observed in DMBA-induced tumor tissues. In addition, in tumor tissues a decrease in lipid order and a significant increase in membrane dynamics were noticed. Further, the composition and secondary structure of proteins were found to be altered, which indicates some important structural alterations in the existing proteins and/or the expression of new types of proteins occurring under the tumor transformation. Furthermore, oral administration of free NAR and NARNPs significantly increased lipids and their order as well as increased the glycogen contents and decreased the levels of proteins and nucleic acid contents. On a comparative basis, NARNPs were found to have a more potent antitumor effect than free NAR in completely preventing the formation of squamous cell carcinoma and in improving the biochemical constituents to a normal range in DMBA-induced HBP carcinogenesis. The present study further shows a great potential of FT-IR spectroscopy as a complimentary tool for the screening of various anticancer drugs and follow-up, which may allow faster response to critical problems arising during treatment.