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Rajiv Gandhi University of Health Sciences, Karnataka Bangalore ANNEXURE II PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION 1 Name Of The Candidate And Address Dr. SYED MOUAZUR RAHAMAN (In Block Letters) POST GRADUATE STUDENT DEPARTMENT OF PERIODONTICS COORG INSTITUTE OF DENTAL SCIENCES, VIRAJPET. 2 Name of the Institution COORG INSTITUTE OF DENTAL SCIENCES, VIRAJPET 3 Course of study and Subject MASTER OF DENTAL SURGERY, PERIODONTICS 4 Date of admission of Course 5 Title of the Topic 26-05-2012 “COMPARATIVE EVALUATION OF MACROPHAGE INFLAMMATORY PROTEIN1α LEVELS IN GINGIVAL CREVICULAR FLUID OF GRADE 0, GRADE I AND GRADE II MOBILE TEETH BEFORE AND AFTER SCALING AND ROOTPLANING IN CHRONIC PERIODONTITIS PATIENTS – A CLINICOBIOCHEMICAL STUDY” Brief Resume of the intended work 6 6.1 Need for the study: Periodontal disease is initiated by plaque, bacteria and perpetuated by the host response to infectious agent residing within plaque biofilm.1 The interaction between oral bacteria and host inflammatory response triggers a cascade of inflammatory events which in turn promote connective tissue destruction and alveolar bone remodeling.2 As early diagnosis can lead to early intervention and prevention of advancing disease, the detection of inflammatory markers and their concentration can be used to develop a test for early clinical diagnosis which could provide early warning of disease and its pathogenesis in advance of clinical and radiographic evidence.1 Gingival crevicular fluid is a rich pool of proteins and molecules that reflect aspects of oral and systemic health. Many studies have concluded that biomarkers of inflammation, connective tissue destruction and bone remodeling were present at elevated levels in GCF of patients with periodontal disease.3 Hence GCF levels of biomarker could be of clinical utility as a screening tool for periodontal disease.2 Macrophage inflammatory protein-1α/CCL3 is a member of the cystine–cystine chemokine family, which is secreted by macrophages,neutrophils, dendritic cells, lymphocytes and epithelial cells.2 Functions of macrophage inflammatory protein-1α include: 1. MIP-1α stimulates monocytes and/or osteoclast progenitor cells to become active osteoclast in dose dependant manner and acts upstream as an activators of osteoclastogenesis within resorption lacunae.1,2 2. It mediates granulocyte migration and adhesion.2 3.It also induces the synthesis of other pro -inflammatory cytokines such as IL-1,IL-6, TNF-alpha from fibroblasts and macrophages.1 Since MIP-1α levels play a significant role in inflammatory events and bone remodeling, there is a need to study its correlation and concentration in GCF with various grades of tooth mobility. 6.2 Review of literature: 1. A longitudinal cohort study of children at risk for aggressive periodontal disease has suggested that MIP-1α could be used as an early biomarker for localized aggressive periodontitis.1,5 2. According to a study MIP-1α has shown to have a powerful link to bone loss in bone resorbing diseases which adds its creditability to be used as a biomarker for identification of patient with risk for bone loss due to periodontal disease.1 3. Studies on salivary biomarkers and gingival crevicular fluid cytokines of periodontal disease in response to treatment has demonstrated reduction in MIP-1α levels following treatment.2,4 4. A study has estimated mean gingival crevicular fluid levels of MIP-1α higher in smokers than in healthy.3 5. A Study has hypothesized that mechanical stress up-regulates the expression of CCL3 in PDL tissues, resulting in chemotaxis and osteoclastogenesis during occlusal traumatism indicating that hyperocclusion stimulates osteclastogenesis via CCL3 expression.7 6. Studies on salivary levels of MIP-1α have concluded that MIP-1alpha has the greatest ability to discriminate periodontal health and disease.2 6.3 Objectives of study: 1. To evaluate GCF concentration of MIP-1α in tooth with grade 0, grade I and grade II mobility of chronic periodontitis patients. 2. To compare GCF concentrations of MIP-1α before and after scaling and root planing in teeth with grade 0, grade I and grade II mobility of chronic periodontitis patients. 3. To correlate the clinical outcome of scaling and root planing with GCF concentrations of MIP-1α in grade 0, grade I and grade II mobile teeth of chronic periodontitis patients. 7 Materials And Methods: 7.1 Source of data: 13 patients reporting to Coorg Institute Of Dental Sciences, Virajpet. 7.2 Method of collection of data: In this study 13 patients (totaling 39 sites) diagnosed of chronic periodontitis will be treated and followed up. 3 sites will be selected randomly from each patient 1.site with PPD≥5mm without tooth mobility(control site) 2.site with PPD≥5mm and with grade I tooth mobility 3.site withPPD≥5mm and with grade II tooth mobility The proposed study will be discussed with the patient and an informed written consent will be taken from the patients. Inclusion criteria Patients aged 35-64yrs Healthy individuals. Patients who can follow the instructions and maintain a good oral hygiene. Exclusion criteria Subjects with any underlying systemic disease Pregnant and lactating women. Tobacco and alcohol users. The use of any drug that may interact or alter MIP-1alpha levels. Subjects who are on any medication or were on any medication since last 6 months. Grade III mobile teeth. Tooth mobility due to trauma Clinical parameters to be used are: Gingival Index (Loe H and Silness J.,1963) Bleeding on probing ( muhlemann & son 1971.,evaluated 30 seconds following pocket probing, a score of 0 (no bleeding) and score of 1 (bleeding) will be given). Clinical Attachment Level (standardized using an acrylic stent and measured by using UNC-15 probe from a fixed reference point to the base of the pocket). Probing Pocket depth (standardized using an acrylic stent and measured by using a UNC-15probe taken from the margin of the gingiva to the base of the pocket). Tooth mobility index(millers mobility index.,1938) no movement distinguishable (0), first distinguishable sign of mobility (1), crown deviates within 1 mm of its normal position (2). All Clinical parameters will be measured at baseline and after 3 months. GCF collection A standard 3 µl of GCF will also be collected using micropipettes after isolation of tooth with cotton rolls from the selected sites at baseline and after 3 months. Collected GCF will be stored at -200C till biomarker analysis is done. Biomarker analysis Concentration of MIP-1α in GCF will be determined using ELISA kit according to manufacturer’s directions. 7.3 Does the study require any investigations or interventions to be conducted on the patients or other humans or animals? ( If so please describe briefly) YES, patients will be informed and GCF collection is done after taking a written consent from the patient. 7.4 Has Ethical clearance been obtained from your institution in case of 7.3? 8 List Of References 1.Fine DH, Markowitz K, Furgang D, Fairlie K, Ferrandiz J, Nasri C, et al. (2009). Macrophage Inflammatory Protein-1alpha: A Salivary Biomarker of Bone Loss in a Longitudinal Cohort Study of Children at Risk for Aggressive Periodontal Disease? Journal of periodontology2009; 80:106113. 2. M. Al-Sabbagh1, A. Alladah1, Y. Lin2, R. J. Kryscio2, M. V. Thomas1, J. L. Ebersole1, C. S. Miller Bone remodeling-associated salivary biomarker MIP-1α distinguishes periodontal disease from health Journal of periodontal research 2012 ;47: 389–395, 3. KA, Guthmiller JM. Influence of smoking on gingival crevicular fluid cytokines in severe chronic periodontitis. Journal of Clinical Periodontology 2011; 38: 219–228. 4.WMSexton, Ylin.Salivary. Biomarkers of periodontal disease in response treatment. JournalofClinical Periodontology 2011;38: 434–441. 5. L.M.shaddox,J.wiedey. Local Inflammatory Markers and Systemic Endotoxin in Aggressive Periodontitis .Journal of dental research September 2011; 90:1140-1144. 6. Thunell DH, Tymkiw KD, Johnson GK, et al. A multiplex immunoassay demonstrates reductions in gingival crevicular fluid cytokines following initial periodontal therapy. Journal of Periodontal research2010;45:148-152. 7. Hyperocclusion Stimulates Osteoclastogenesis via CCL2 Expression K.T. Goto,H. Kajiya,Department of Dental Hygiene, Fukuoka College of Health Sciences, Fukuoka 8140193, Japan.journal of dental research 2011; 90: 793-798. to 9. Signature of the Candidate 10. Remarks of the Guide 11. Name & Designation of (In block letters) 11.1 Guide DR.B.S.JAGADISH PAI 11.2 Signature 11.3 Co- guide (if any) 11.4 Signature 11.5 Head of the Department DR.PADMARAJAN (PROFESSOR & HOD) 11.6 Signature 12. 12.1 Remarks of the chairman & Principal DR. SEQUEIRA PETER SIMON (PRINCIPAL) 12.2 Signature