Screening of Oral Squamous Cell Carcinoma Through Color Intensity-Based Textural Features.

Background Early screening and diagnosis of oral squamous cell carcinoma (OSCC) has always been a major challenge for pathologists. Artificial intelligence (AI)-assisted screening tools can serve as an adjunct for the objective interpretation of Papanicolaou (PAP)-stained oral smears. Aim This study aimed to develop a handy and sensitive computer-assisted AI tool based on color-intensity textural features to be applied to cytologic images for screening and diagnosis of OSCC. Methodology The study included two groups consisting of 80 OSCC subjects and 80 control groups. PAP-stained smears were collected from both groups. The smears were analyzed in Matlab software computed data and color intensity-based textural features such as entropy, contrast, energy, homogeneity, and correlation, were quantitatively extracted. Results In this study, a statistically significant difference was noted for entropy, energy, correlation, contrast, and homogeneity. It was found that entropy and contrast were found to be higher with a decrease in homogeneity, correlation, and energy in OSCC when compared to the control group. Receiver operating characteristic curve analysis was done and accuracy, sensitivity, and specificity were found to be 88%, 91%, and 81%, respectively. Conclusion The gray-level co-occurrence matrix (GLCM) color intensity-based textural features play a significant role in differentiating dysplastic and normal cells in the diagnosis of OSCC. Computer-aided textural analysis has the potential to aid in the early detection of oral cancer, which can lead to improved clinical outcomes.

Significance of HIF-1α and CD105 in establishing oral squamous cell carcinoma associated with oral submucous fibrosis a distinct clinicopathological entity.

Context: Oral squamous cell carcinoma associated with oral submucous fibrosis (OSCC with OSMF) is clinicopathologically a distinct entity. However, scientific proof in view of assessment of biomarkers of hypoxia and neoangiogenesis to differentiate them are lacking. The expression of hypoxia-inducible factor 1-α (HIF-1α) and CD105 in OSCC with and without OSMF possibly will be explicated along these lines. Aim: This study aims to evaluate the molecular basis of hypoxia and neoangiogenesis in terms of immunohistochemical expression of HIF-1α and CD105 in OSCC with and without OSMF cases. Settings and Design: A retrospective cohort. Subjects and Methods: The study comprise of 203 histopathologically diagnosed surgically operated cases of OSCC retrieved from the departmental archives. The OSCC cases were subgrouped into two, OSCC with OSMF (Group I) and OSCC without OSMF (Group II). The evaluation of hypoxia and angiogenesis was carried out by immunohistochemical markers, HIF-1α and CD105. MVD is the parameter of angiogenesis expressed by CD105. Statistical Analysis Used: Differences in CD105, and HIF-1α immunoreactivity between study groups were done using descriptive statistics using "Kruskal-Wallis test," "Mann-Whitney test." Statistical significance was set at P < 0.05. Results: On comparison of MVD in Group I and II, statistically significant difference was found in MVD (8.88 ± 3.41, 16.13 ± 5.86, P = 0.0001). The HIF1-α expression was less in Group I (6.85 ± 2.62) as compare to Group II (7.22 ± 3.08) but the difference was statistically nonsignificant (P = 0.35). Conclusions: The OSCC with OSMF is not only clinicopathologically distinct entity of OSCC but also diverse in its molecular pathogenesis as explicited by distinct expression of HIF-1 α and CD105.

Odontogenic tumors: A review of 93 cases in the Vidharba region of Maharashtra.

BACKGROUND: Tumors of the odontogenic origin (OTs) are rare tumors accounting for 1% of all the jaw tumors in the oral cavity. The behavior of these tumors ranges from hamartomatous proliferation to malignant neoplasm. AIM: The aim of this study was to determine the epidemiological data, clinical and histopathological picture with variants of the OTs diagnosed at our institute in the Vidarbha region. SETTINGS AND DESIGN: A retrospective study was carried out with the permission from the institutional authorities. The archival records of the Department of Oral Pathology and Microbiology were reviewed. MATERIALS AND METHODS: A total of 93 cases of OTs were reviewed retrospectively from the archival records of the Department of Oral Pathology and Microbiology from January 2008 to December 2018. Statistical analysis was carried out and the determination of the age, gender, jaw, site, diagnosis, variants and year-wise distribution of odontogenic lesions was taken out using the SPSS software. RESULTS: Ninety-three cases of OTs were identified. The most common OT identified was ameloblastoma (n = 37/39.7%) followed by keratocystic OT (n = 8/8.6%), adenomatoid OT (n = 7/7.5%) and odontome (O) (n = 6/6.4%). The tumors were diagnosed in a wide age range from 1st to 3rd decennium of life. The mandibular posterior region was the most commonly affected anatomical site with the maxilla and mandible ratio of 1:6. CONCLUSION: OTs were found to be rare in the sample studied. The findings of the study were in concordance to those of Asian and African series. Variations were shown from the series of American reports, and further investigations are needed for this disparity.

Oral submucous fibrosis: An enigmatic morpho-insight.

Oral submucous fibrosis (OSMF) is a chronic progressive, scarring disease affecting oral, oropharyngeal, and sometimes the esophageal mucosa. It is characterized by the progressive fibrosis of the submucosal tissue. The pathogenesis of OSMF has been directly related to the habit of chewing areca nut and its commercial preparation, which is widespread in Indian subcontinent and Southeast Asia. The areca nut has been classified as a "group one human carcinogen." Oral squamous cell carcinoma in the background of OSMF is one of the most common malignancies in South and Southeast Asian countries. Malignant transformation has been reported in 7%-12% cases of OSMF. Histopathological spectrum of OSMF includes the apparent alterations observed in the epithelium and connective tissue. Epithelial atrophy and sometimes epithelial hyperplasia with or without dysplasia are the peculiar alterations seen in the epithelium. In the connective tissue, there is extracellular matrix remodeling which results in excessive collagenization. Further cross-linking of collagen leads to hyalinization which makes the collagen resistant to proteolysis. Owing to fibrosis in the connective tissue, there is narrowing of blood vessels which further results in compromised blood supply to the local tissue milieu, that is, hypoxia. This tissue hypoxia elicits angiogenesis which may result in the malignant transformation of OSMF. Perpetual irritation of areca nut and its constituents to the oral mucosa leads to upregulation of pro-inflammatory cytokines and further juxtaepithelial inflammation. Thus, these coordinated reactions in epithelium and connective tissue leads the OSMF toward malignant transformation.

Ki67, CD105, and α-SMA expression supports the transformation relevant dysplastic features in the atrophic epithelium of oral submucous fibrosis.

BACKGROUND: The grading of oral epithelial dysplasia is not possible in the atrophic epithelium of oral submucous fibrosis (OSMF). Recently, we found that features such as increased basal cell layer hyperplasia, abnormal superficial mitosis, increased nuclear-cytoplasmic ratio, increased nuclear size, and hyperchromasia represent transformation-relevant dysplastic features in the atrophic epithelium of OSMF. The presence of these features can be considered a high-risk feature for patients. However, these findings have not been tested and authenticated using markers relevant to oral carcinogenesis. METHOD: Paraffin-embedded tissues from 30 normal oral mucosa (NOM) and 50 OSMF were retrieved from 2008 to 2016 and subjected to immunohistochemical expression using Ki67, CD105 and α-SMA antibodies. RESULTS: Ki67 LI showed significant increases from NOM (12.47±2.34) to LRED (23.47±3.75) to HRED (34.31±7.31) (<0.0001). Similarly, MVD was increased significantly from NOM (3.53±5.17) to LRED (27.57±12.25) to HRED (46.18±12.55) (p<0.0001). The expression of α-SMA was significantly increased from LRED (0.21±0.41) to HRED (1.13±0.56) (<0.0001). The Ki67 LI and α-SMA; MVD and α-SMA; and Ki67Ki67 LI and MVD in NOM, LRED and HRED showed a statistically significant positive correlation (P<0.0001). The increase in Ki67 LI was directly proportional to MVD and α-SMA expression from NOM to LRED to HRED (P<0.0001). The connective tissue stroma of NOM lacks α-SMA expression. Mild myofibroblast expression was noted in 4 cases of LRED (14.28%) and in 18 cases of HRED (81.81%). Moderate expression was noted only in 4 cases of HRED (22.22%). CONCLUSION: Ki67 LI, CD105, and α-SMA expression showed significant differences between normal, LRED and HRED. These findings further support that features such as increased basal cell layer hyperplasia, abnormal superficial mitosis, increased nuclear-cytoplasmic ratio, and hyperchromasia could be transformation-relevant dysplastic features.

Impact of virtual microscopy with conventional microscopy on student learning in dental histology.

BACKGROUND: In dental histology, the assimilation of histological features of different dental hard and soft tissues is done by conventional microscopy. This traditional method of learning prevents the students from screening the entire slide and change of magnification. To address these drawbacks, modification in conventional microscopy has evolved and become motivation for changing the learning tool. Virtual microscopy is the technique in which there is complete digitization of the microscopic glass slide, which can be analyzed on a computer. This research is designed to evaluate the effectiveness of virtual microscopy with conventional microscopy on student learning in dental histology. MATERIALS AND METHODS: A cohort of 105 students were included and randomized into three groups: A, B, and C. Group A students studied the microscopic features of oral histologic lesions by conventional microscopy, Group B by virtual microscopy, and Group C by both conventional and virtual microscopy. The students' understanding of the subject was evaluated by a prepared questionnaire. RESULTS: The effectiveness of the study designs on knowledge gains and satisfaction levels was assessed by statistical assessment of differences in mean test scores. The difference in score between Groups A, B, and C at pre- and post-test was highly significant. This enhanced understanding of the subject may be due to benefits of using virtual microscopy in teaching histology. CONCLUSION: The augmentation of conventional microscopy with virtual microscopy shows enhancement of the understanding of the subject as compared to the use of conventional microscopy and virtual microscopy alone.

Early Stage Oral Submucous Fibrosis is Characterized by Increased Vascularity as Opposed to Advanced Stages.

INTRODUCTION: The degree of vascularity of the diseased mucosa in Oral Submucous Fibrosis (OSMF) has always been a matter of debate with conflicting results. Knowledge of this aspect is important to understand pathogenesis of OSMF, which in future could be translated into therapeutic strategies. AIM: In the present study, attempt has been made to investigate parameters like Mean Vascular Density (MVD), Total Vascular Area (TVA) and Mean Vascular Area (MVA) using CD34 antibody. MATERIALS AND METHODS: Forty five previously untreated histopathologically diagnosed cases of OSMF were retrieved from archives and fifteen age and sex matched healthy volunteers without habits were included in the control group. Sections were immunohistochemically stained for CD 34 and morphometric analysis was performed. For statistical analysis ANOVA, Kruskal Wallis test and Mann Whitney U tests were used and p-values <0.05 were considered statistically significant. RESULTS: MVD was more in Stage I OSMF followed by Control, Stage II and Stage III with statistically significant differences (p< 0.001). Statistically significant differences were observed in the MVD between control versus Stage III OSMF. Similarly, TVA was statistically significant when compared between control versus OSMF, control versus Stage II OSMF, control versus Stage III OSMF, Stage I versus Stage II OSMF, Stage I versus Stage III OSMF, and Stage II versus Stage III OSMF. For MVA, significant differences were between control versus OSMF, control versus Stage II OSMF, control versus Stage III OSMF, Stage I versus Stage III OSMF and Stage II versus Stage III OSMF. CONCLUSION: Angiogenesis is seen in early stages of OSMF with decreasing trend in advanced stages. Decreased vascular areas seen in advanced stages could be attributed to the increasing fibrosis surrounding the blood vessels.

Oral squamous cell carcinoma in the background of oral submucous fibrosis is a distinct clinicopathological entity with better prognosis.

BACKGROUND: The aim of this study was to compare the clinicopathological features of oral squamous cell carcinoma in the background of oral submucous fibrosis (OSCC-OSMF) and oral squamous cell carcinoma (OSCC). METHODS: A total of 217 cases of OSCC were retrieved from achieves for the analysis. OSCC-OSMF cases were segregated on the basis of history and clinicopathological parameters. RESULTS: The study included 217 patients of which 112 had OSCC and 105 OSCC-OSMF. OSCC-OSMFs were younger compared with OSCC. Overall oral cancer was noted predominantly in males compared to females. The number of OSCC-OSMF was more in clinical TNM stage I and stage II as compared to OSCC, whereas the number of OSCC was more in stage III and stage IV compared to OSCC-OSMF. Histological presentation of well-differentiated squamous cell carcinoma was significantly more in OSCC-OSMF compared to OSCC, whereas moderately differentiated squamous cell carcinoma was significantly more in OSCC compared to OSCC-OSMF. Regional lymph node metastasis was significantly higher in OSCC compared to OSCC-OSMF. Three-year disease-free survival rate was significantly higher in OSCC-OSMF compared to OSCC. CONCLUSION: The OSCC-OSMF was found to be a clinicopathologically distinct entity with a better grade of tumor differentiation, less incidence of nodal metastases, and early detection (early clinical TNM stage) compared to OSCC. All these factors probably contribute to a better prognosis and increased 3-year disease-free survival in OSCC-OSMF patients.

Matrix metalloproteinases-3 gene-promoter polymorphism as a risk factor in oral submucous fibrosis in an Indian population: a pilot study.

AIM: Host-related factors could be important to the fundamental understanding of oral diseases. One such factor is matrix metalloproteinases (MMP). MMP3 (stromelysin 1) is crucial in connective tissue remodeling. Promoter regions are specific segments of DNA that control the rate of mRNA synthesis. Thus, polymorphism in the (5A/6A) promoter region of the MMP3 gene results in different transcriptional activities and is related to susceptibility in diseases. The aim of the present study was to detect polymorphism in the MMP3 gene-promoter region in oral submucous fibrosis patients (OSF), oral squamous cell carcinoma patients (OSCC), and normal individuals with tobacco and areca nut habit and without lesions, and to correlate the genotype as a risk factor in these diseases. METHOD: Genomic DNA from blood of OSF (n = 5), OSCC (n = 5), and normal individuals with tobacco and areca nut habit (n = 5) and without (n = 5) were subjected to polymerase chain reaction of the MMP3 gene-promoter region and DNA sequencing. RESULTS: The 5A allele in the MMP3 gene-promoter region was observed more frequently in the OSF group than the control groups. No significant difference was noted between OSCC and the control groups on the 5A allele. CONCLUSION: The results indicated that the 5A allele of the MMP3 gene-promoter region could be associated with OSF risk factor, but not OSCC, in an Indian population.

Evaluation of Effect of Astringent on Oral Mucosa as a Non-surgical Preprosthetic Treatment Modality in Edentulous Patients: An In Vivo Study.

Preprosthetic treatments are advocated in edentulous patients to enhance the denture bearing areas for good denture support. Most of the times the preprosthetic treatments are considered only in a surgical way. Ideally every edentulous patient undergoing complete denture treatment needs a non-surgical preprosthetic treatment. So that, the denture bearing area will be properly prepared before the denture construction. The present study was conducted on thirty completely edentulous male patients who had visited to our Institute for the treatment. Each patient was asked to massage with astringent gel on the denture bearing mucosa over a 4 weeks period. Exfoliative cytology was used to collect the surface cells from the palatal mucosa. First scrape was taken before the stimulation treatment was started. The second and third scrape was taken after the stimulation treatment with astringent gel for each patient. In this way total 90 scrapes were made and the each smear was stained with the Papanicolaou's technique to examine under light microscope. About 100 cells were counted from each stained smear. The number of parabasal cells, intermediate cells and superficial cells were recorded to calculate the degree of keratinization. Statistical analysis was performed. A significant difference (p < 0.001) in keratinization levels was found. The mean percentage of superficial cells before the stimulation treatment was 79.80 %; this percentage was gradually increased to 84.60 and 90.57 % after the 2  and 4 weeks period of stimulation treatment respectively with astringent gel.

Evaluation of chemiluminescence, toluidine blue and histopathology for detection of high risk oral precancerous lesions: A cross-sectional study.

BACKGROUND: Early detection holds the key to an effective control of cancers in general and of oral cancers in particular. However, screening procedures for oral cancer are not straightforward due to procedural requirements as well as feasibility issues, especially in resource-limited countries. METHODS: We conducted a cross-sectional study to compare the performance of chemiluminescence, toluidine blue and histopathology for detection of high-risk precancerous oral lesions. We evaluated 99 lesions from 55 patients who underwent chemiluminescence and toluidine blue tests along with biopsy and histopathological examination. We studied inter-as well as intra-rater agreement in the histopathological evaluation and then using latent class modeling, we estimated the operating characteristics of these tests in the absence of a reference standard test. RESULTS: There was a weak inter-rater agreement (kappa < 0.15) as well as a weak intra-rater reproducibility (Pearson's r = 0.28, intra-class correlation rho = 0.03) in the histopathological evaluation of potentially high-risk precancerous lesions. When compared to histopathology, chemiluminescence and toluidine blue retention had a sensitivity of 1.00 and 0.59, respectively and a specificity of 0.01 and 0.79, respectively. However, latent class analysis indicated a low sensitivity (0.37) and high specificity (0.90) of histopathological evaluation. Toluidine blue had a near perfect high sensitivity and specificity for detection of high-risk lesions. CONCLUSION: In our study, there was variability in the histopathological evaluation of oral precancerous lesions. Our results indicate that toluidine blue retention test may be better suited than chemiluminescence to detect high-risk oral precancerous lesions in a high-prevalence and low-resource setting like India.