Apoptotic and mitotic indices in oral epithelial dysplasia and squamous cell carcinoma: A comparative study.

Background: Assessment of apoptotic cells and mitotic figures using light microscopy is an easy and viable alternative to assess tumour behaviour. Aims: To evaluate apoptotic index (AI), mitotic index (MI) and apoptotic to mitotic index ratio (AI: MI) in different grades of oral epithelial dysplasia (OED) and oral squamous cell carcinoma (OSCC) in haematoxylin and eosin-stained (H&E) sections. Settings and Design: The study included 45 cases each of OED and OSCC cases which were further subgrouped into groups of 15 each based on their grades. AI, MI and AI/MI were assessed and compared with 15 cases of normal mucosa. Methods and Material: Apoptotic cells and mitotic figures were counted using a binocular light microscope equipped with an oculometer grid (20 × 20 squares) on the eyepiece. Cells were counted in 15 grid fields under oil immersion lenses (×100) in a stepladder fashion. AI/MI ratio was calculated. Statistical Analysis: The results obtained were statistically analysed using Analysis of variance and Tukey Honestly Significant Difference tests with SPSS 20 software at a 0.05 significance level. Results: AI increased with increasing grades of dysplasia and decreased with increasing grades of OSCC. MI increased with increasing grades of OED and OSCC. AI/MI increased with increasing grades of OED but decreased with increasing grades of OSCC. Conclusion: In the light of the current observations, AI, MI and AI: MI can be considered as valuable parameters to assess the biological behaviour of OED and OSCC.

Cytomorphologic Analysis of Wet and Spray Fixation Methods in Oral Exfoliative Cytology

@#For a long time, oral exfoliative cytology (OEC) has been implemented as an effective preliminary diagnostic tool for pathological lesions and various methods for fixation of the cytology specimens have been studied. The present study was undertaken to compare the efficacy between the wet and spray type of fixation methods for Papanicolaou (PAP) stained oral cytosmears. The study comprised of 45 healthy subjects in the age group of 20-25 yrs. For each subject, two smears were collected from the buccal mucosa and subjected to wet and spray fixation methods respectively. Both the smears were stained using a commercial Rapid Pap Kit. Smears were observed microscopically and evaluated for cytomorphological features involving uniformity of staining, cellular morphology, nuclear morphology, cellular staining, nuclear staining and presence of impurities. Comparisons were made between the two methods of fixation and statistically analysed using McNemar non-parametric test. Cells were evenly distributed in wet-fixed smears (n=38, 95%) compared to spray fixed smears (n=19, 47.5%). Wet-fixed smears showed lesser impurities (n=13, 32.5%) than spray fixed smears (n=27, 67.5%). However, other parameters such as cytological and nuclear morphology, staining of cytoplasm and nucleus were found to be not significant when compared between the two methods of fixation (p<0.05). The study shows that wet-fixed smears have better cellular distribution and relatively fewer impurities when compared to the spray fixed smears. The method of wet-fixed smears may be used as an alternative to spray fixed smears. A larger sample size may be required for further validation.

Speciation of Candida using CHROMagar in cases with oral epithelial dysplasia and squamous cell carcinoma.

BACKGROUND: Candida albicans is most frequently isolated from oral cavity but identification of other Candida species such as C. tropicalis, C. krusei, C. glabrata & C. dubliniensis is increasing proportionately. A constant rise in immuno-suppressed patients, widening range of recognized pathogens, and resistance to antifungal drugs are contributing factors which stress the need for species identification of Candida, an opportunistic pathogen. Objectives: 1. To detect the prevalence of Candida albicans and Non albicans Candida albicans (NAC) species in the oral cavity of patients with epithelial dysplasia, Oral squamous cell carcinoma (OSCC) and healthy controls. 2. To identify and differentiate Candidal species using CHROMagar, a differential media. MATERIAL AND METHODS: The study included smears from 50 patients with histopathological confirmation of epithelial dysplasia & OSCC and 50 normal controls. Candida albicans was identified using Sabouraud dextrose agar media (SDA) as primary culture followed by species identification using CHROMagar on the basis of colony color and morphology. RESULTS: Non albicans candida predominated (66%) over Candida albicans (34%) in speciation on CHROMagar media in the study group. Non albicans Candida species isolated were C. tropicalis (38%), C. glabrata (24%) and 2 cases showing polyfungal population of C. albicans & C. glabrata. CONCLUSIONS: Species level isolation of Candida helps in early identification of resistant non Candida strains and prompt treatment of the cases there by preventing the dissemination of infection in case of immuno-compromised individuals. The data presented also supports the use of CHROMagar Candida as a pertinent media for the rapid identification of Candida species directly from clinical specimens in resource challenged settings, which could be helpful in developing appropriate therapeutic strategy and management of patients. Key words:Candida, CHROMagar, epithelial dysplasia, oral cancer.

Oral rinse-based cytology and conventional exfoliative cytology: a comparative study.

CONTEXT: Simple screening test for resource-challenged areas. AIMS: To compare specimen adequacy and diagnostic agreement between oral rinse-based smears and conventional smears in oral cancer, oral leukoplakia, and normal controls. SETTINGS AND DESIGN: Twenty-five histologically confirmed cases of oral cancer, 29 clinically diagnosed cases of oral leukoplakia, and 54 normal controls were subjected to the oral rub-and-rinse technique and the conventional exfoliative cytology. PATIENTS AND METHODS: Smears were consequently stained with the Papanicolaou (PAP) stain and were examined independently by two different cytopathologists in a double-blind fashion. The results were classified as: Negative, atypical, positive, and unsatisfactory. Furthermore, they were compared, to assess the sample adequacy, cellular distribution, staining, leucocytes/inflammation and cellular clarity in both the smears. STATISTICAL ANALYSIS USED: Chi square test and Wilcoxon-matched pairs signed rank test (P ≤ 0.05). RESULTS: Among the cancer cases, 84% showed positive, 8% atypical, and 8% unsatisfactory, after using the conventional technique, whereas, the oral rinse technique showed 80% positive, 16% atypical, and 4% unsatisfactory cases. The conventional technique showed 44.82, 37.93, and 6.89% and the oral rinse technique showed 48.27, 31.03, and 20.68% negative, positive, and atypical cases, respectively, among the leukoplakia cases. Among the normal controls, 9.26 and 90.74% and with the oral rinse technique 3.70 and 96.29% showed atypical and negative cases with the conventional technique. Sample adequacy and cellular clarity showed a statistically significant difference between both techniques. CONCLUSIONS: Both smears were diagnostically reliable, however, the oral rinse-based method showed an overall improvement in sample adequacy and cellular clarity.

Oral rinse as a simpler approach to exfoliative cytology: a comparative study.

BACKGROUND: Oral rinse is a novel method that can be used to detect dysplasia in potentially malignant disorders and malignant oral lesions in resource challenged areas. A study was undertaken to compare the quality of the normal smears prepared with the oral rinse and that of the wooden tongue spatula. MATERIAL AND METHODS: One hundred five normal subjects were selected for the study. Two smears were prepared from clinically normal mucosa using an oral rinse and further two smears were scraped from clinically normal buccal mucosa using a wooden spatula. STATISTICAL ANALYSIS: The smears were graded for cell yield, dispersion and cellular clarity on a three-point scale by two observers. The results were analyzed using Mann Whitney non parametric test. RESULTS: The Oral rinse was found to be significantly more efficient than the wooden spatula, in terms of cell yield (p<0.0001), cell dispersion (p =.0052) and cellular detail (p<0.0001). CONCLUSION: The study showed that the Oral rinse is an effective method for use in exfoliative cytology of normal oral mucosa.