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Title:	Expression of collagenase, proinflammatory cytokines and enterococcus faecalis gene in the periodontitis-induced rats: an indicative association between periodontitis with carditis and nephritis
Authors:	Fazle Khuda (P94735)
Supervisor:	Nurrul Shaqinah Nasruddin, Dr.
Keywords:	Periodontal Diseases Gingivitis Periodontitis Universiti Kebangsaan Malaysia -- Dissertations Dissertations, Academic -- Malaysia
Issue Date:	26-Apr-2022
Description:	Periodontal disease is an inflammatory disease in which microbial aetiological factors trigger a cascade of host responses that mediate inflammatory events. The disease is initiated as gingivitis and if untreated can progress to periodontitis. It is suggested that inflammation caused by periodontal disease may be responsible for the risk of heart disease and kidney disease. Although presence of Enterococcus faecalis in the healthy oral cavity is low or rare but its association with endodontic lesion as well as periodontitis has been identified. Dysregulation of the collagenase and released of proinflammatory cytokines appear to have a key role in periodontal tissue destruction. Thus, the aim of the study is to determine the expression pattern of collagenase, proinflammatory cytokines and E. faecalis gene in several organs of inducedperiodontitis rats. Rats were divided into three groups containing four rats each for 0- day (control), 7-day and 14-day. Sterile ligature wire 0.2 mm was inserted into the interdental space of maxillary right 1st and 2nd molars and 0.5 ul of E. faecalis suspension of 1.5x108 CFU/ml was injected into the gingival sulcus between maxillary right 1st and 2nd molar area once a week during respective induction period. Following euthanasia, tissue samples from gingiva, heart and kidney from all rats were collected for RNA extraction, cDNA conversion and gene amplification of collagenase (MMP-1, MMP-8, MMP-13) and proinflammatory cytokines (IL-1β, IL-6, TNF-𝛼) using quantitative Real-Time PCR. The samples were further used for DNA extraction and quantification of E. faecalis bacterial gene by RT-qPCR. At the same time, maxilla, heart and kidney samples were collected, fixed, embedded and proceed for H&E staining. The RT-qPCR results showed significant mRNA expression of MMP-1 (p<0.05), MMP-8, -13 (p<0.01) at 7-day within the gingival tissue sample, as compared control group which decreased at 14-day. All collagenase expressions downregulated in the heart and only MMP-1 and MMP-13 were upregulated in the kidney at 7- and 14-day post infection. The expression levels of proinflammatory cytokines within the gingival tissue, heart and kidney were upregulated at both 7- and 14-day as compared to control group. E. faecalis gene expression showed significant upregulation in heart and kidney at both 7 and 14-day post infection. Histological analysis of maxilla showed apical migration of the junctional epithelium, loss of attachment from cementoenamel junction, inflammatory cell infiltration and presence of osteoclasts as well as bone resorption within the periodontium of 7-day group, and lesion severity were lessened in 14-day group as compared to 7-day group. In the heart, inflammatory cell infiltration within the myocardium has observed as well. Congested blood vessel within the glomerulus and inflammatory cells infiltration into the interstitial spaces were also observed in the kidney samples. The results suggest acute inflammatory response following infection and wire placement which suggested possible association between periodontitis with carditis and nephritis. Hence the study will contribute to future research on the identification of potential oral biomarkers in the early diagnosis of periodontitis and associated disease,Ijazah Sarjana Sains Pergigian
Pages:	111
Publisher:	UKM, Kuala Lumpur
Appears in Collections:	Faculty of Dentistry / Fakulti Pergigian

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