Evaluation of <em>Candida</em> species in oral cavities of patients with oral submucous fibrosis and healthy controls: A comparative study 

The objective of the study was to find out the prevalence of oral squamous cell carcinoma (OSCC) with oral submucous fibrosis (OSF) in patients with OSF. Of 48,757 patients, we found 300 OSF subjects. Three hundred patients of OSF were checked for OSCC. Both OSF and OSCC with OSF (OSCCwOSF) were diagnosed histopathologically. The prevalence of OSCCwOSF was calculated. Descriptive analysis was done. Chi-square test and t-test were calculated for proportions and mean, respectively, to check any difference among OSF and OSCCwOSF groups. Age-specific relative risk was calculated in OSF and OSCCwOSF groups. Multiple logistic regression analysis was done among odd ratios of the different variable between OSF and OSCCwOSF groups. The prevalence of OSCCwOSF among OSF was 13.7% over a period of 1 year. The mean age of OSCCwOSF group was 43.95 ± 10.22 years in comparison to the OSF group that was 35.51 ± 11.26 years (P < 0.00). The mean habit duration was significantly less in the OSF group when compared to OSCCwOSFgroup for mishri (P = 0.002). Age-specific adjusted relative risk of OSCC in OSF patient increases from 0.33 (18-34 years) to 3.86 (≥65 years). It could be concluded that a 13.7% prevalence rate of OSCCwOSF in OSF patients should alert the clinician. Clinicians should, therefore, anticipate OSSC in OSF patients. This awareness could lead to the early diagnosis and management of such OSCC.

Background and Objectives:Oral submucous fibrosis (OSF) is extensively prevalent in India and South-East Asia owing to the habit of arecanut (AN) use. Epithelial atrophy, hyposalivation, and immune alterations in OSF may predispose to increased Candidal carriage. Stomatopyrosis in OSF can result from multiple causes, which may also include Candidal infection. Hence, this study is aimed to assess Candidal carriage, species characterization, salivary flow rate (SFR) and its relationship with the clinical features (stomatopyrosis and mouth opening [MO]) in OSF patients alongwith the response to antifungal treatment in patients with higher Candidal carriage.Methodology:In this case-control study, 60 OSF patients and 30 age- and sex-matched control subjects were enrolled. SFR was assessed using modified Schirmer test. Samples for Candidal assessment were collected with the oral rinse technique and cultured. The isolated yeast species were counted and identified based on Gram staining, germ tube test, and CHROMagar. Data were analyzed with Chi-square test, Pearson's correlation test, and one-way ANOVA test.Results:The distribution of mean visual analog scale (VAS) score, SFR and MO was significantly varied (P < 0.001) in the study and control groups. Candida was found to be present significantly (P = 0.048) in OSF group as compared to control group. Candida albicans was the predominant species. No statistically significant association was obtained regarding Candidal isolation and SFR, burning sensation and MO in OSF patients. Only 1 patient in the study group yielded a high Candidal carriage (>400 CFU/mL) and reported relief in burning sensation (VAS score) with antifungal therapy.Conclusions:OSF patients yielded a significant higher oral Candidal carriage. Although it was not found to be associated directly, its role as a “cause and effect” in SFR and clinical features (stomatopyrosis and MO) of OSF cannot be ignored.

Hematinic deficiencies and anemia in gastric parietal cell antibody-positive and -negative oral submucous fibrosis patients

This study aimed to correlate the oxidative stress marker levels in saliva with the clinical stage based on mouth opening, fibrotic bands and histopathological grades of oral submucous fibrosis (OSF) patients. The study included patients clinically diagnosed with OSF (n=63) and equal number of age and gender matched controls. Patients with OSF were defined by mouth opening stage, fibrotic bands and histopathological grades. Unstimulated saliva from both control and OSF patients were analysed for oxidative markers like lipid peroxides (LPO), non-enzymic antioxidants [reduced glutathione (GSH), vitamin A, vitamin E, vitamin C] and enzymatic antioxidants [glutathione peroxidase (GPx), superoxide dismutase (SOD)] and correlated with different stages and grades. Total salivary protein and LPO were significantly increased in OSF group with no significant change in the levels of GSH compared to controls. In OSF patients, a significant decrease in the levels of vitamins A, C and E was observed. The activities of salivary SOD and GPx were significantly decreased in OSF patients compared to controls. These changes significantly correlated with the increasing and differing grades of OSF that reflects increased oxidative stress with the progress of OSF.

Aim:To investigate the presence of lysyl oxidase (LOX) G473A polymorphism in group 1 {Oral submucous fibrosis (OSMF) patients}, group 2 (betel quid chewers without OSMF) and group 3 (healthy individuals).Materials and Methods:A total of 60 patients were taken for the study, which included 20 OSMF patients (group 1), 20 betel quid chewers without OSMF (group 2) and 20 healthy individuals without OSMF and betel quid chewing habit (group 3). DNA was isolated using Qiagen kit. The isolated DNA was quantified using spectroscopic methods. Polymerase chain reaction (PCR) was carried out at annealing temperature of 67o C. PCR amplification was checked on 2% agarose gel. Further, the amplified PCR products were subjected to automated DNA sequencer, to assess LOX G473A polymorphism.Results:The gene sequence data generated from the automated DNA sequencer was received as colored electropherograms. These gene-sequencing results did not show LOX G473A polymorphism in any of the 3 groups.Conclusion:In our study, gene-sequencing results did not show LOX G473A polymorphism in OSMF patients. Since only one study in the literature has shown the association of LOX gene polymorphism and OSMF patients, we conclude that further studies are required to unveil the role of LOX gene polymorphism in OSMF.

Estimation and correlation of plasma fibrinogen degradation product (FDP) and salivary fibrin precipitating factor (FPF) in oral sub mucous fibrosis (OSMF) patients, betel quid chewers without OSMF and healthy individuals. The study aims to evaluate whether FDP and FPF can be used as a marker for development and progression of OSMF and whether there is any correlation between the two. The study included 163 subjects grouped into three categories, Group 1, which included 54 control patients, Group 2, which included 55 betel quid chewers, and Group 3, which included 54 clinically confirmed OSMF patients. All of them were subjected to the estimation of plasma FDP and salivary FPF. FDP was present in 52 (96.3%) patients in Group 3, 2 (3.6%) patients in Group 2, and 1 patient in Group 1 (1.9%). FPF was positive in eight patients. The correlation of FDP and FPF was done by Fisher's exact test and was found to be statistically nonsignificant. In the present study, FDP was positive in 52 (96.3%) OSMF patients; hence, FDP may be used as an early indicator of OSMF. FDP was found to be positive in two patients with a habit of chewing betel quid without OSMF. It may be hypothesized that these patients are more likely to develop OSMF. Hence, the estimation of FDP may be used as a diagnostic test to predict an impending OSMF before it could manifest itself clinically. FPF was negative in some OSMF patients, may be because it is below the detectable range.

Oral Submucous Fibrosis (OSMF) is a chronic debilitating disease more frequently encountered in the South-East Asian population. This disease represents a public health priority as it is grouped within oral potentially malignant disorders, with malignant transformation rates of around 7–19%. Hence, early identification of high-risk OSMF patients is of the utmost importance to prevent malignant transformation. Among various biomarkers, EGFR overexpression has an unfavorable clinical outcome, poor prognosis, and low survival rates in Oral Squamous Cell Carcinoma (OSCC). The current study aimed to evaluate the expression of EGFR in saliva and exfoliated buccal cells of OSMF. Immunoexpression of EGFR was observed in healthy controls (n = 11), OSCC (n = 106), and OPMD with dysplasia (n = 56), which showed significant expression with increasing grades of dysplasia and OSCC. EGFR expression was evaluated in saliva and exfoliated buccal cells of healthy controls (n = 15), OSMF (n = 24), and OSCC (n = 10) patients using ELISA, which revealed significant expression in OSMF and OSCC. Validation studies were also performed using real-time PCR (RT-PCR) to compare gene expression in healthy controls (n = 9), OSMF (n = 9), and OSCC (n = 25), which showed significant 18-fold upregulation in OSCC and three-fold upregulation in OSMF when compared to healthy controls. Hence, saliva and exfoliated buccal cells could be considered as potential non-invasive diagnostic samples for the evaluation of high-risk patients of OSMF using EGFR as a biomarker.

Objective: To determine the maximum bite force (MBF) in oral submucous fibrosis (OSMF) patients and to compare them with that of healthy subjects. Material and Methods: Twenty patients who were clinically confirmed, as OSMF and 20 healthy controls matched for age, gender, and number of intact functional teeth were included in this study. For each subject, age, gender, weight, height and body mass index (BMI) were recorded. The MBF registration was carried out by the two evaluators, who were previously calibrated. Bite force was measured in the first molar region using a force transducer occlusal force meter for each subject seated at the upright position, with Frankfort's plane nearly parallel to the floor, and no head support. The Student’s independent t-test was used to determine the statistical significance in relation to mean height, weight, BMI and the presence of number of intact teeth and MBF between the healthy subjects and OSMF individuals. A comparison of grades of OSMF with all variables was carried out by one-way ANOVA test. Results: No significant difference was found in mean age, mean height, weight, BMI and the presence of the number of intact teeth between healthy individuals and OSMF patients. The mean MBF in healthy subjects was 628.23 ± 24.39 N and 635.47 ± 31.22 N in OSMF patients. Even though the healthy subjects reported a higher MBF than OSMF patients did, the difference was statistically non-significant. With regards to sides, no significant difference was observed in mean MBF in healthy subjects and OSMF patients on the right (p=0.7818) and left side (p=0.6154). Conclusion: The healthy subjects reported higher MBF values than OSMF patients did and the difference was statistically non-significant.

Objectives: The increased muscle activity and diminished blood supply following connective tissue changes in Oral Submucous Fibrosis (OSF) patients lead to muscle degeneration and fibrosis. The histopathological examination of OSF patients confirms fibrosis, but it also causes scarring and worsens OSF. Ultrasonography (USG) has been utilized in recent research to assess the echogenic patterns of the head and neck muscles in OSF patients. The study aimed to compare the echogenic pattern of the masseter muscle in OSF patients to that of normal mucosa using USG. Methods: The study population was split into cases and controls. The cases were 63 subjects who were clinically diagnosed with OSF. In comparison, the controls were 63 subjects who were randomly selected from the outpatient clinic of Oral Medicine and Radiology, Government Dental College, Thiruvananthapuram. According to Khanna and Andrade’s criteria, OSF subjects were divided into four groups based on their clinical findings. A high-resolution real-time LOGIQUE C5 ultrasonic scanner with 7-12MHz transducers was used to image the masseteric muscle. Results: In the present study, type II internal echogenic pattern (64.4 %) was predominant, followed by type III and type I in OSF subjects. So, type I internal echogenic patterns might be associated with normal muscles, and types II and III internal echogenic patterns might be associated with pathologic changes. A good interobserver agreement was observed. Conclusion: In OSF patients, the appearance of the masseter muscle’s internal echogenic pattern showed a varied pattern from that of normal types.

Purpose: Oral submucous fibrosis (OSMF) is a high-risk precancerous condition that predominantly affects Indian youngsters due to the habit of gutkha chewing. Candida may play a role in the etiopathogenesis of premalignant and malignant lesions. The aim of this study is to compare the incidence, intensity, and species of Candida found in OSMF patients and healthy individuals. Methods: This study included 20 OSMF patients and 20 healthy controls. A detailed history of each patient was recorded along with a clinical examination. Samples were collected with the oral rinse technique and cultured on Sabouraud’s agar medium. The isolated yeast species were counted and identified based on Gram staining, a germ tube test, chlamydospore formation and a sugar assimilation test. Results: In total, 40% of OSMF patients and 15% of healthy controls yielded Candida organisms on culture. C. albicans was the predominant species isolated, but C. krusei and C. tropicalis were also identified. Gender, gutkha habit and clinical staging had no influence on the candidal carriage in OSMF patients. Conclusion: The incidence and intensity of Candida (primarily C. albicans) was greater in OSMF patients than in healthy controls, but these findings were within the normal limit (3-47%). Therefore, Candida may not be an etiologic factor in malignant transformation. However, controversy still exists over whether the chewing of betel quid in cases of OSMF has an inhibitory effect or promotes the adherence and invasion of Candida.

Aims: The present clinico-radiological study was done to evaluate the morphological variants of soft palate in oral submucous fibrosis (OSMF) patients using digital lateral cephalometry. Different variations in the morphology of soft palate were compared with stages of OSMF. Further, soft palate morphology in OSMF patients was compared radiographically with that of normal population. Materials and Methods: A total number of 100 patients who were a part of this study were divided in two equal Groups. Group 1 comprised of 50 patients clinically diagnosed with OSMF and Group 2 included 50 routine patients. Results: Six different morphological variants of soft palate were found. Among the study Groups, type 1soft palate was most commonly seen (56%) whereas type 5 was the least common variant. Majority of patients belonged to stage II OSMF and type 1soft palate was commonly seen in this stage of disease whereas butt shaped soft palate (type 3) was more common in stage III OSMF. Conclusion: In OSMF, type 1 and 2 are commonly seen but as the diseases advances, these are replaced by type 3 and 6 variants. In OSMF patients, there in reduction in the anterio-posterior dimension of soft palate.

Morphometric evaluation of soft palate in oral submucous fibrosis—A digital cephalometric study

Background: Oral submucous fibrosis (OSMF) is a chronic premalignant condition seen in the oral cavity, but sometimes extends to the pharynx, esophagus, or even the larynx. OSMF causes stiffness in the oral cavity leads to trismus and affects speech and swallowing. Sometimes this lesion affects the Eustachian tube opening at the nasopharynx which results in Eustachian tube dysfunction followed by middle ear diseases. The purpose of the present work is to evaluate otological manifestations in OSMF patients. Materials and Methods: Patients with OSMF were evaluated for otological manifestations such as hearing impairment and fullness in the ear. All of them were evaluated with pure tone audiometry and tympanometry. Results: In this study group of 58 patients of OSMF comprising 116 ears, the hearing was within the normal limit in 60 ears (51.72%), a mild degree of conductive deafness in 52 ears (44.82%), a mild degree of sensorineural deafness in 2 ears (1.72%) and a moderate degree of sensorineural deafness were present in 2 ears (1.72%). There is a significant difference in mean quantitative hearing impairment was seen in OSMF groups C and group D with respect to the control group. Conclusion: Involvements of palatal muscles in OSMF patients reduce the patency of the Eustachian tube, resulting in a conductive type of deafness. OSMF patients should be assessed for hearing impairment and suggested for proper management.

Introduction: Ultrasonography (USG) serves as a valuable, non-invasive tool to monitor these changes, providing insights into the muscle’s structure and function. The normal thickness of the masseter muscle differs by gender, with males generally having thicker muscles both at rest and during contraction compared to females. Oral submucous fibrosis (OSMF), a potentially malignant disorder, significantly increases the risk of oral cancers, particularly squamous cell carcinoma. Aims and objective: To evaluate the difference in cross-sectional thickness and internal echogenic pattern of masseter muscle at rest and at maximum clenching position using ultrasonography in chronic areca nut /tobacco chewers, OSMF patients and control groups. Materials and methods: The study participants were categorized into three groups with 75 partcipants ,consisting of 25 participants each with the following conditions; Group 1 with chronic chewers, group 2 with OSMF and group 3 with healthy individuals as controls. An ultrasonographic examination of masseter was performed in all subjects for masseter muscle thickness and echogenic pattern, the echogenic pattern was classified into Types I, II and III. Differences in masseter muscle thickness and echogenic pattern between study groups and control group was evaluated. Results: the age group were between 20 and 50 years of age with significant male predominance. The thickness of the masseter muscle was significantly greater in chronic chewers, both in the relaxed and contracted states, on both sides, when compared to both the OSMF group and the control group. In contrast, the OSMF group exhibited a decrease in masseter muscle thickness compared to the control group. Conclusion: The findings revealed a statistically significant increase in masseter muscle thickness in chronic chewers, both in the relaxed and contracted states, on both sides, when compared to both OSMF patients and the control group. A slight muscle atrophy that is decrease in the muscle thickness was observed in OSMF patients in comparison to the control group

Aim: To examine and compare cytomorphometric changes in the keratinocytes obtained from buccal mucosa of oral submucous fibrosis (OSF) cases in different clinical stages with healthy subjects and oral squamous cell carcinoma (OSCC) cases. Materials and Methods: Buccal mucosal smears of OSF cases [ n = 30], OSCC cases [ n = 30] and age-sex matched healthy controls [ n = 30] were stained with Papanicolou stain and observed under light microscope (40x objective magnification) and 100 cells in each case were analysed cytomorphometrically using an image analyser. The average nuclear area (NA), cell area (CA), ratio of nuclear area/cell area, nuclear diameter (ND), cell diameter (CD) and ratio of nuclear diameter/cell diameter was obtained for each case. One-way ANOVA and Tukey-HSD procedure ( P &lt; 0.05) were used to analyze all the parametric variables. Results: The comparison of mean values of the NA, CA, NA:CA, ND, CD, ND:CD shows statistically significant difference among the healthy, OSF and OSCC groups [ P &lt; 0.05]. OSF group showed significant increase in nuclear dimensions and N/C ratio and significant decrease in cell dimensions compared to healthy group ( P &lt; 0.05). Among the parameters analysed ND:CD and NA:CA showed a highly statistically significant difference between the OSF and healthy group ( P &lt; 0.001). Conclusions: There are significant alterations in the cell and nuclear dimensions of the buccal mucosal cells in OSF patients. These alterations could be attributed to usage of various tobacco and areca nut preparations and due to the mucosal changes (atrophic epithelium, high epithelial turnover) in the OSF. Atrophic epithelium, in OSF is envisaged to predispose malignant transformation with persistent exposure oral carcinogens. As the mean values of CA, NA:CA in stage IV OSF are close to OSCC cases, it strengthens the view that advancement of fibrosis increases the risk of development of epithelial dysplasia.