Orthokeratinised odontogenic cyst: A case series.

Orthokeratinised odontogenic cyst (OOC) is a distinct and an uncommon odontogenic cyst of jaw. It was previously considered as orthokeratinized variant of Odontogenic Keratocyst, owing to its same site, age and occurrence and origin. In 2017 WHO classified OOC under odontogenic cysts as a distinct entity, clearly separating it from Odontogenic Keratocyst (OKC). In cases of malignant transformation, the cyst shows unclear boundaries. The cases that reported to the department during the period 2019-2021 were retrieved and then reviewed by the authors. A total of 7 cases that met the histopathological criteria for OOC were included in the study. There was a clear male predilection. Age of patients ranged between 17 to 65 years. Most of cases showed Unilocular radiolucency but one case revealed Multilocular pattern. Grossly specimens ranged between 0.5 cm to 3.0cms. Histopathological findings showed all classic features of OOC. FNAC was also done and it is clearly evident that the use of Cytology can help us diagnose the cyst early and is a less invasive method. OOC is a rare type of odontogenic cyst, but while making the diagnosis one should not forget about the fact that it is less aggressive and needs to be differentiated from OKC. Majority times it has been seen that Clinico-radiographic diagnosis given of any circumscribed radiolucency around an impacted tooth is Dentigerous cyst, which is not correct. Diagnosing such lesions require thorough histopathological and radiographical correlation along with molecular analysis to reach final diagnosis.

Candidal carriage in diabetic patients: a microbiological study.

OBJECTIVE: Candida yeast species are widespread opportunistic microbes and incidence of its carriage in diabetic patients compared with non-diabetic controls is not clearly defined. The present study was carried out to isolate and identify the candida species in type I, type II diabetes mellitus patients and in controls. Study comprised of 60 subjects in the age range of 20 to 65 years, consisting of two groups of diabetes patients with controlled diabetic status (20 subjects each with type I and type II) and 20 age and sex matched controls. Saliva samples were collected from all groups and inoculated on Sabouraud's Dextrose Agar (SDA) to check for the fungus growth. The confirmed candidal colonies were further inoculated on CHROMagar for different colour differentiation. Out of 60 samples, 40 (66.67%) showed a positive fungal prevalence. Out of 40 positive fungal prevalence 82.50% were representative of candida. The positive fungal prevalence for candida I was 48.48%, in group II was 30.30%, in group III was 21.21 %. Further speciation in CHROMagar revealed different species of candida predominantly of candida albicans 20% and few mixed culture of candida albicans with candida krusei 5% and candida albicans with candida tropicalis 5% in group I. In group II, candida albicans was 35% and mixed culture of candida albicans with candida glabrata was 5%. In group III, candida albicans was 25% and candida tropicalis was 5%. The findings confirm that diabetic patients harbor yeast in their oral cavity and are more susceptible to oral candidiasis and also that CHROMagar medium is a satisfactory isolation medium for oral cavity specimens, allowing rapid and accurate identification of yeast colonies with easy recognition of mixed culture and is easy to use.

Prevalence, knowledge, and attitude of gutkha chewing among school children of Arsikere, India.

AIMS: To assess the prevalence, knowledge, and attitude of gutkha chewing among school children of Arsikere, India. MATERIALS AND METHODS: Two thousand school children aged between 10 and 15 years were examined with individual interviews along with a questionnaire to evaluate the presence of betel nut and paan chewing habit in various forms. Children of both sexes were included in the study. Responses of all study population and the association between dependent and explanatory variables were assessed using Chi-square test. RESULTS: Twenty-eight percent of children had the habit of gutkha chewing, more among boys than girls at the ratio of 4:1 which was statistically significant (P < 0.05). The habit was more common in government school children than private school children; over 70% of children believed that it is a bad habit, but half the study population was not aware of side effects; 30% of children believed that it is not a bad habit and has no side effects. CONCLUSION: There is a higher prevalence of gutkha chewing habit in school children who are not aware of the side effects. Children themselves purchase the gutkha and betel nut sachets, hence the Government should ban the sale and purchase by children.

Effect of staining procedures on the results of micronucleus assay in the exfoliated buccal mucosal cells of smokers and nonsmokers: A pilot study.

BACKGROUND: Tobacco consumption in smoke form causes severe health problems such as cancer. Micronuclei are structures that present after genomic damage in buccal mucosal cells which could be a good indicator of chromosomal alterations in cytological samples. The nuclear anomalies that are consequences of cell injury are also found in these cells. AIM: The present study was aimed at evaluating the effectiveness of deoxyribonucleic acid (DNA)-specific stain over DNA-nonspecific stain in exfoliated buccal mucosa cells in smokers and nonsmokers for assessment of micronuclei and nuclear anomalies. MATERIALS AND METHODS: Thirty subjects were divided into three groups; Group 1 (control) nonsmokers (n = 10), Group 2 individuals with history of smoking for less than 10 years (n = 10), and Group 3 individuals with history of smoking for more than 10 years (n = 10). The exfoliated buccal mucosal cells were scrapped using wooden spatula, smeared over the glass slide, and stained with DNA-nonspecific stain, Giemsa; and DNA-specific stains, Feulgen and acridine orange. RESULTS: We found that the mean micronuclei score with DNA-nonspecific stain, Giemsa, was significantly higher in smokers group when compared to nonsmokers group. No such significant differences were observed with DNA-specific stains. Also, in the smokers groups, mean micronuclei were higher in Group 3 when compared to Group 2. CONCLUSION: The score of MN assay and nuclear anomalies in exfoliated buccal mucosal cells of smokers and nonsmokers strongly depended on staining procedures and duration of smoking. Hence, these should be interpreted with caution to avoid false-positive results.

Spritzer: For diagnostic cytopathology.

BACKGROUND: Exfoliative cytology is an easy, economical, noninvasive, and feasible method for early detection and screening program of any premalignant or malignant lesion. In case of routine cytological procedure, classical Papanicolaou (PAP) stain is widely used while Romanowsky stains are sparingly used. Leishman-Giemsa (LG) cocktail, being a easier, cost effective staining technique, has not been used in exfoliative cytology. Therefore, this pilot study was carried out to compare and contrast the role of LG stain in routine cytological procedure which is very cost-effective, less time-consuming and requires less infrastructural support. AIM AND OBJECTIVES: To study the diagnostic efficiency of LG cocktail in comparison with PAP stain and Feulgen stain in mucosal cells for evaluating cellular changes of oral squamous cell carcinoma (OSCC). MATERIALS AND METHODS: Three cytological smears were prepared from 10 healthy controls and 10 patients clinically diagnosed with OSCC, and they were stained with LG cocktail stain, PAP stain, and Feulgen stain. The stained smears were evaluated for cytologic diagnosis and the staining characteristics such as nuclear and cytoplasmic details were recorded as per criteria by Sujathan et al. STATISTICAL ANALYSIS: The data were statistically evaluated with analysis of variance test using SPSS 15 software for windows. RESULTS: The results from the cases diagnosed as OSCC by PAP and LG cocktail were almost identical and superior to Feulgen stain both in diagnostic ability and in staining characteristics. CONCLUSION: The one-step LG cocktail is easy, very cost-effective, less time-consuming with less infrastructural support as compared to PAP stain; however, it warrants further evaluation for screening of oral cancer as a potential aid.

An update on studies on etiological factors, disease progression, and malignant transformation in oral submucous fibrosis.

Worldwide estimates of oral submucous fibrosis (OSMF) show a confinement to Indians and Southeast Asians. In India, the prevalence of OSMF has increased over the past four decades from 0.03% to 6.42%. The condition is well recognized for its malignant potential rate of 7-30%. The condition has a multifactorial etiology and may remain either stationary or become severe, leaving an individual physically challenged both physically and psychologically. Hence, the study aims at reviewing studies done on various etiological factors leading to its onset. Their analysis may serve as an adjunct in defining the broad spectrum of the causation of this potentially malignant disorder.

Spindle cell lesions: A review on immunohistochemical markers.

The intriguing array of spindle cell lesions occurring, especially in the head and neck region, poses a critical diagnostic challenge not only to the histopathologist but also ultimately to the clinicians for planning an appropriate treatment protocol. Overlapping spectrum of clinico-radiographic and microscopic features further compounds this problem. In such situations, the aid of ancillary techniques such as immunohistochemistry (IHC) is sought to clinch the diagnosis. This study aims to review the spindle cell neoplasms of the oral cavity with emphasis on IHC.

A study of the distribution of B-cell lymphoma/leukemia-2 in odontogenic cyst and tumors: Histochemical study.

INTRODUCTION: The odontogenic keratocyst is known for its aggressiveness, high recurrence rate, and transformation of keratinized epithelia to nonkeratinized squamous epithelium for which inflammation has been suggested to be responsible. B-cell lymphoma/leukemia-2 (Bcl-2) an anti-apoptotic protein, prolongs the life span of epithelial cells and allows proliferation, differentiation, and morphogenesis. MATERIALS AND METHODS: Study was carried out comprising of 90 cases; (30 ameloblastoma, 30 keratocystic odontogenic tumor (KCOT), and 30 radicular cyst). Bcl-2 expression was determined with respect to localization, area (percentage) and intensity of stained cells in epithelium, and connective tissue stroma by counting the endothelial, round, and fusiform cells. RESULTS: In epithelium bcl-2 expression in KCOTs was higher followed by ameloblastoma and lowest in the radicular cyst. Whereas, in connective tissue stroma bcl-2 expression was higher in KCOT and radicular cyst than ameloblastoma cases. Solid variants showed statistically higher expression as compared to the unicystic variants of ameloblastoma (P = 0.009, 0.033, 0.011, and 0.041). CONCLUSION: High expression of bcl-2 in KCOT supports the general agreement that some features of KCOT are those of a neoplasia. The bcl-2 expression in connective tissue cells suggests that these cells may also be important as epithelial cells in the biological behavior odontogenic keratocyst.

Oxidative stress marker in oral cancer: A review.

Oxygen derived species such as hydrogen peroxide, superoxide anion radical, hydroxyl radical (OH-), and singlet oxygen are well known to be cytotoxic and have been implicated in the etiology of a wide array of human diseases, including cancer. Various carcinogens may also partly exert their effect by generating reactive oxygen species (ROS) during their metabolism. Oxidative damage to cellular DNA can lead to mutations and may, therefore, play an important role in the initiation and progression of multistage carcinogenesis. ROS influences central cellular processes such as proliferation, apoptosis, and senescence which are implicated in the development of cancer. Understanding the role of ROS as key mediators in signaling cascades may provide various opportunities for pharmacological intervention.

Immunohistochemical expression of vascular endothelial growth factor in keratocystic odontogenic tumor, dentigerous cyst, and radicular cyst: A comparative study.

BACKGROUND: Cyst and tumors arise from tissue remains of odontogenesis, these interactions have been considered to play an important role in the tumorigenesis of odontogenic lesions. The connective tissue stroma has an essential role in the preservation of epithelial tissues and minor alterations in the epithelium are followed by corresponding changes in the stroma, such as angiogenesis. Vascular endothelial growth factor (VEGF) is considered the first factor which maintains its position as the most critical driver of vascular formation and is required to initiate the formation of immature vessels, with this aim, present study was executed to evaluate VEGF expression in kertocystic odontogenic tumor, dentigerous cyst and radicular cyst (RC). MATERIALS AND METHODS: A retrospective study was carried out comprising a total of 31 cases; 13 cases of keratocystic odontogenic tumor (KCOT), nine cases of dentigerous cyst (DC) and nine cases of RC. The sections were stained immunohistochemically with VEGF antibody and were evaluated for the presence and intensity of the immuno reactive cells. Statistical analysis was carried out using Chi-square test to inter-compare the VEGF expression between KCOT, DC, and RC. RESULTS: VEGF expression in the epithelium and connective tissue was significantly higher in KCOT compared to dentigerous and RC. One case of KCOT with carcinomatous change also revealed positive results for the VEGF expression in the dysplastic epithelium, tumor islands, and connective tissue. The significant difference was observed on inter-comparison of the VEGF expression in the connective tissue of KCOT and DC, whereas no significant difference was observed in the VEGF expression in the connective tissue of KCOT and DC. CONCLUSION: The present study data supports the literature finding that angiogenesis can be important in the progression and enlargement of odontogenic cysts similarly to what occurs in neoplastic conditions and further it can be concluded that the higher positivity for VEGF of KCOT could help to explain in part the aggressive biological behavior of the lesion. The stroma of KCOT could be regarded not only as a structural support of the cyst wall but also as playing a part in the neoplastic behavior of cyst.

Evaluation of the Secretor Status of ABO Blood Group Antigens in Saliva among Southern Rajasthan Population Using Absorption Inhibition Method.

INTRODUCTION: The ABO blood group system was the significant element for forensic serological examination of blood and body fluids in the past before the wide adaptation of DNA typing. A significant proportion of individuals (80%) are secretors, meaning that antigens present in the blood are also found in other body fluids such as saliva. Absorption inhibition is one such method that works by reducing strength of an antiserum based on type and amount of antigen present in the stains. AIM: To check the efficacy of identifying the blood group antigens in saliva and to know the secretor status using absorption inhibition method among southern Rajasthan population. MATERIALS AND METHODS: Blood and saliva samples were collected from 80 individuals comprising 20 individuals in each blood group. The absorption inhibition method was used to determine the blood group antigens in the saliva and then the results were correlated with the blood group of the collected blood sample. The compiled data was statistically analysed using chi-square test. RESULTS: Blood groups A & O revealed 100% secretor status for both males and females. While blood groups B and AB revealed 95% secretor status. CONCLUSION: Secretor status evaluation of the ABO blood group antigen in saliva using absorption inhibition method can be a useful tool in forensic examination.

Estimation of salivary lactate dehydrogenase in oral leukoplakia and oral squamous cell carcinoma: a biochemical study.

BACKGROUND: Enzyme Lactate Dehydrogenase (LDH) is found in the cells of almost all body tissues. The profile of salivary total LDH enzymes is similar to that found in oral epithelium, indicating that the major source of salivary LDH is probably the oral epithelium-shedding cells. Consequently, LDH concentration in saliva, as an expression of cellular necrosis, could be a specific indicator for oral lesions that affect the integrity of the oral mucosa. MATERIALS AND METHODS: Study comprised of three groups as follows: Group I: Comprised of 25 healthy individuals of comparable age. Group II: 25 otherwise healthy and consenting patients with oral leukoplakia (OL). Group III: 25 otherwise healthy and consenting oral squamous cell carcinoma (OSCC) patients. Biochemical estimation of LDH was done with the help of Semiautomatic Analyzer. RESULTS: Inter comparison of salivary total LDH levels between all the three groups revealed that salivary LDH levels increase from healthy control group to Oral Leukoplakia group to further increase in OSCC group. On comparisons between the histopathological grades of OSCC group the level of LDH were found to increase from well differentiated to moderately differentiated to further increase in poorly differentiated patients. CONCLUSION: The present salivary analysis for LDH enzyme reveals an overall altered salivary LDH enzyme level in OL and OSCC cases.

Role of bacteria in oral carcinogenesis.

Oral cancer appears to be increasing in incidence, and mortality has hardly improved over the past 25 years. Better understanding of the etiopathogenesis should lead to more accurate and earlier diagnosis and more effective treatments with fewer adverse effects. Despite increasing interest in the possible relationships between bacteria and the different stages of cancer development, the association of bacteria with cancer of the oral cavity has yet to be adequately examined. Different bacteria have been proposed to induce carcinogenesis, either through induction of chronic inflammation or by interference, either directly or indirectly, with eukaryotic cell cycle and signaling pathways or by metabolism of potentially carcinogenic substances like acetaldehyde, causing mutagenesis. This review presents the possible carcinogenesis pathway involved in bacterial carcinogenesis, commonly implicated bacteria in oral carcinogenesis and their role in cancer therapeutics as well.

Awareness of General Dental Practitioners about Oral Screening and Biopsy Procedures in Udaipur, India.

PURPOSE: Oral cancer presents with high mortality rates, and the likelihood of survival is remarkably better when detected early. The present study aimed to assess the awareness of general dental practitioners (GDPs) about oral screening and biopsy procedures in Udaipur, India. MATERIALS AND METHODS: In this cross-sectional study, 83 GDPs were surveyed using a self-administered structured questionnaire consisting of several mandatory and optional questions. The data were analysed and frequency distribution was performed. RESULTS: Most of the GDPs adequately performed complete oral cavity examinations and were aware of suspicious oral lesions, most common sites and risk factors for oral pre-cancer/cancer, but did not inquire about patients' tobacco/ alcohol consumption habits. Half of them referred lesions requiring biopsy to a specialist/higher centre rather than performing biopsies themselves, even after recognising the importance of biopsy as a diagnostic tool due to concerns of inadequate experience and instruments required. Varied results regarding selection of the appropriate site for biopsy and preservation of biopsied specimens were noted. CONCLUSION: Most of the GDPs were adequately aware of oral screening and biopsy procedures but felt reluctant to perform them, which suggests that dental education programmes are needed for GDPs in oral pre-cancer/cancer detection as well as screening and diagnostic procedures.

Nuclear anomalies in exfoliated buccal epithelial cells of petrol station attendants in Udaipur, Rajasthan.

AIM: The petroleum derivatives consist of a complex mixture of chemical compounds one among which is benzene. Petrol station workers who pump fuel to vehicles absorb the products of fuel fumes and the products of combustion. MATERIALS AND METHODS: To study the occupational exposure to petroleum derivatives such as benzene, exfoliated buccal cells from 40 petrol station attendants and 40 age-matched control subjects were examined for micronuclei, binucleation, karyorrhexis and karyolysis frequency by using feulgen and Giemsa stains. Statistical evaluation was performed with ANOVA test. RESULTS: In the present study, inter-comparison of mean values for micronuclei, binucleation, karyorrhexis and karyolysis using Fuelgen stain between smokers of study and control group, as well as between smokers and non-smokers of study and control group revealed statistically highly significant results with P value 0.00002 and 0.0001 respectively. Whereas inter comparison between non-smokers of study and control group using Feulgen stain and inter comparison between smokers of study and control group, using Giemsa stain revealed statistically significant results with P value 0.0034 and 0.0004, respectively. CONCLUSION: This study revealed that the studied individuals belong to a risk group and should periodically undergo biological monitoring and proper care.

Evaluation of stromal myofibroblasts in oral leukoplakia, oral submucous fibrosis, and oral squamous cell carcinoma--an immunohistochemical study.

BACKGROUND: Oral leukoplakia (OL) and oral submucous fibrosis (OSMF) are the main potentially malignant disorders and oral squamous cell carcinoma (OSCC) is the most common malignancy of the oral mucosa. Myofibroblasts (MFs) secrete numerous growth factors and inflammatory mediators that stimulate epithelial cell proliferation and play an important role in tumoral invasion and use a combination of different factors in the course of neoplastic growth and development. Hence the present study was undertaken to evaluate and compare the distribution of MFs using alpha smooth muscle actin (α-SMA) in OL, OSMF, and various histopathological grades OSCC. MATERIALS AND METHODS: Sixty formalin-fixed paraffin-embedded tissue blocks consisting of histopathologically diagnosed cases of normal mucosa (n = 10), OL (n = 14) hyperkeratosis with various grades of dysplasia, OSMF (n = 11), and OSCC (n = 25) were subjected to immunohistochemistry using α-SMA antibody for detection of MFs. RESULTS: MFs were not detected in normal oral mucosa. On comparison of frequency of mean scores in OL, OSMF, and OSCC the values were 0.6 ± 0.2 (0-2), 1.2 ± 0.68 (1-2), and 2.6 ± 1.34 (0-4), respectively. The results were statistically significant (P < 0.001). CONCLUSION: These findings are suggestive of role of MFs with the creation of a permissive environment for tumor invasion in OSCC. Hence the presence of MF is a prognostic marker and evaluation of the frequency in the stroma can be used as therapeutic targets.

Evaluation of salivary and serum lipid peroxidation, and glutathione in oral leukoplakia and oral squamous cell carcinoma.

Lipid peroxidation induced by reactive oxygen species (ROS) is involved in the pathogenesis of malignancy. Overall, lipid peroxidation levels are indicated by malondialdehyde (MDA), which is the most frequently used biomarker to detect oxidative changes. Antioxidant defense systems such as glutathione (GSH) limit cell injury induced by ROS. Therefore, MDA and GSH can be used to monitor oxidative stress (OS). Hence, this study aimed to evaluate and compare both salivary and serum levels of MDA and GSH in oral leukoplakia and oral squamous cell carcinoma (OSCC) patients, and healthy controls. The study included 100 subjects comprising 30 apparently healthy controls, 30 patients with oral leukoplakia and 40 clinically and histologically diagnosed patients with OSCC. Saliva and blood samples were obtained and evaluated for MDA and GSH. The study revealed enhanced MDA levels in saliva and serum in oral leukoplakia and OSCC patients as compared to controls. On the other hand, significant decreases were seen in serum and salivary GSH levels in oral leukoplakia and OSCC patients as compared to controls. Augmentation of OS in blood and saliva is reflected by increase in MDA and decrease in GSH levels, indicating that tumor processes cause an imbalance of oxidant-antioxidant status in cell structures.

Calretinin as a diagnostic adjunct for ameloblastoma.

Background. Calretinin is a 29 kDa calcium-binding protein of the EF-hand family which is expressed in a variety of normal and tumorigenic tissues. Its expression in odontogenic epithelium during odontogenesis and in neoplastic odontogenic tissues has been demonstrated. Unicystic ameloblastoma poses a diagnostic challenge, as its histologic presentation can be sometimes mistaken for keratocystic odontogenic tumor (KCOT). This study was performed to assess the usefulness of calretinin as a confirmatory marker for ameloblastic tissue. Methodology. Total of 40 cases: 16 unicystic ameloblastoma, 4 multicystic ameloblastoma, and 20 KCOT, were evaluated immunohistochemically for the presence, localization, distribution, and intensity of calretinin expression. Statistical analysis was done using Chi-square test to intercompare the expression between ameloblastoma and KCOT. Results. Sixteen cases of ameloblastoma (12 unicystic, 4 multicystic) showed positive calretinin staining of ameloblastic epithelium and only one case of KCOT was positive for calretinin, with the positivity restricted to the stellate reticulum like epithelium. Intercomparison between two groups revealed statistically significant difference (P = 0.000). Conclusion. Calretinin appears to be a specific immunohistochemical marker for neoplastic ameloblastic epithelium and may be an important diagnostic adjunct in the differential diagnosis of ameloblastoma and KCOT.

Expression of cell cycle and apoptosis-related proteins in ameloblastoma and keratocystic odontogenic tumor.

Tumors arising from epithelium of the odontogenic apparatus or from its derivatives or remnants exhibit considerable histologic variation and are classified into several benign and malignant entities. A high proliferative activity of the odontogenic epithelium in ameloblastoma (AM) and keratocystic odontogenic tumor (KCOT) has been demonstrated in some studies individually. However, very few previous studies have simultaneously evaluated cell proliferation and apoptotic indexes in AM and KCOT, comparing both lesions. The aim of this study was to assess and compare cell proliferation and apoptotic rates between these two tumors. Specimens of 15 solid AM and 15 KCOT were evaluated. The proliferation index (PI) was assessed by immunohistochemical detection of Ki-67 and the apoptotic index (AI) by methyl green-pyronin stain. KCOT presented a higher PI than AM (P < .05). No statistically significant difference was found in the AI between AM and KCOT. PI and AI were higher in the peripheral cells of AM and respectively in the suprabasal and superficial layers of KCOT. In conclusion, KCOT showed a higher cell proliferation than AM and the AI was similar between these tumors. These findings reinforce the classification of KCOT as an odontogenic tumor and should contribute to its aggressive clinical behavior.

Hidden keys in stroma: Unlocking the tumor progression.

Malignancy is considered as a pathological imbalance of tissue-cell societies, a state that emerges from tumor-host microenvironment in which host participates in induction, selection and expansion of the neoplastic cells. Invasion of these malignancies can be viewed as a derangement in the proper sorting of cell populations, causing a violation of normal tissue boundaries. This violation is carried out by certain stromal cells like carcinoma associated fibroblasts (CAFs), tumor associated macrophage (TAMs), endothelial cells (ECs) leucocytes, bone marrow derived cells, etc. Tumor cells may alter the surrounding stroma and in turn, stromal cells may promote cancer progression and invasion. Thus, this review compares the role of CAFs, TAMs and ECs in tumor microenvironment towards tumor progression. This compilation aims to collate existing literature on stromal cell with particular emphasis on their role in tumor invasiveness and summarize experimental studies, trials and literature of last 10 years collected from pubmed central indexed journals.