Rebecca Jason, (2022) Comparative Analysis of Localized and Systemic Genotoxicity in Smokers With and Without Lesion using Micro Nucle1 Assay: A Case Control study. Masters thesis, Rajas Dental College and Hospital, Tirunelveli.
Full text not available from this repository.Abstract
BACKGROUND: Development of tobacco smoking associated lesions are a well-documented phenomenon and the potential of these lesions turning malignant make them a health hazard. The presence of micronuclei has been established as a reliable marker for determining genetic damage. Therefore, evaluation of genotoxicity in smokers with (case) and without (controls) tobacco smoking associated lesions can be performed using micronuclei assay (Mn) in both exfoliated buccal mucosal cells as well as in in- vivo mitogen stimulated peripheral blood lymphocytes. AIM OFTHE STUDY: To evaluate and compare genotoxicity using micronuclei assay in exfoliated buccal mucosal cells and peripheral blood lymphocytes in smokers with (case) and without (control) tobacco smoking associated lesions. OBJECTIVE: To evaluate and compare micronuclei (MN) frequency in exfoliated buccal mucosal cells among smokers with and without tobacco smoking associated lesions and then to compare MN frequency in peripheral blood lymphocytes among smokers with and without tobacco smoking associated lesions. HYPOTHESIS: There is an increased localized and systemic genotoxicity (i.e. increased frequency of MN) in cases than in controls. METHODOLOGY: A case control study was designed with 15 individuals with lesions (cases) and 15 individuals without lesions (controls). Buccal mucosal micronuclei assay: Buccal mucosal cells were collected by rubbing the buccal mucosa with a wooden spatula and directly spreading the smear onto a clean glass slide and fixed in Carnoys fixative. The smears were later stained using DNA specific Feulgen stain and counter stained with Light green. Cytokinesis- block micronuclei assay: 5 ml of heparinized blood was obtained and added to culture media containing 5ml RPMI 1640, 1ml FBS and 0.3 ml phytohemagglutinin and incubated at 37˚C for 44 hours. At the end of 44th hour cytochalasin-B is added to block the mitosis and the culture is reincubated for 24 hours. Cells are harvested between the 72nd and 74th hour from initial incubation. Cell pellets are made and used to prepare smears which are stained using Geimsa stain. RESULTS: There was an increase in micronuclei in the peripheral blood lymphocytes and buccal mucosal cells of cases as compared with controls indicating a higher localized and systemic genotoxicity in cases. Corelating the micronuclei in buccal exfoliated cells with peripheral blood lymphocytes, a statistically significant increase was observed in peripheral blood lymphocytes of both cases and controls which indicates a higher systemic genotoxicity. CONCLUSION: Based on the results of this study we see that a higher genotoxicity occurs in individuals with smoking associated oral lesions. This genotoxicity is evident both locally and systematically. Therefore, micronuclei assay can serve as an early indicator of development of oral malignancies.
| Item Type: | Thesis (Masters) |
|---|---|
| Additional Information: | 241921551 |
| Uncontrolled Keywords: | Micronuclei, Cytokinesis-block micronuclei assay, Buccal mucosal micronuclei assay, smoking associated lesions, genotoxicity. |
| Subjects: | DENTAL > Oral Pathology and Microbiology |
| Depositing User: | Subramani R |
| Date Deposited: | 08 May 2022 09:13 |
| Last Modified: | 14 May 2022 07:36 |
| URI: | http://repository-tnmgrmu.ac.in/id/eprint/19788 |
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