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RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES BANGALORE, KARNATAKA. ANNEXURE II 1. NAME OF THE CANDIDATE AND ADDRESS DR. GARIMA GARG POST GRADUATE STUDENT DEPARTMENT OF PERIODONTICS GOVERNMENT DENTAL COLLEGE AND RESEARCH INSTITUTE, BANGALORE. 2. NAME OF THE INSTITUTE GOVERNMENT DENTAL COLLEGE AND RESEARCH INSTITUTE, BANGALORE. 3. COURSE OF STUDY AND SUBJECT MASTER OF DENTAL SURGERY IN PERIODONTICS. 4. DATE OF ADMISSION TO COURSE 01.06.2007 5. TITLE OF THE TOPIC: “CORRELATION OF THE LEVELS OF CATHEPSIN K IN GINGIVAL CREVICULAR FLUID AND SERUM IN PERIODONTAL HEALTH, DISEASE AND AFTER TREATMENT-A CLINICO-BIOCHEMICAL STUDY.” 6. BRIEF RESUME OF THE INTENDED WORK: 6.1 NEED FOR THE STUDY: Periodontal diseases are a complex group of diseases characterized by inflammation and destruction of tooth-supporting tissue. Proteinases are among the mediators produced as a part of host response that contribute to tissue destruction. Cathepsin K is an acidic cysteine endoproteinase, abundantly but not exclusively expressed in osteoclasts. It plays a critical role in bone remodeling by degradation of bone matrix (collagen type I, type II and osteonectin). Cathepsin K reactivity is observed in osteoclasts located in the resorption lacunae and multinuclear giant cells.1 Levels of cathepsin K are found to be significantly higher in serum of patients with longstanding inflammatory disease, rheumatoid arthritis.2 Increased cathepsin K mRNA is detected in mononuclear and multinuclear osteoclasts on the pressure side of the alveolar bone of rat after orthodontic force application.3 Elevated levels of cathepsin K are reported in gingival crevicular fluid (GCF) in patients with periodontitis4 and peri-implantitis.5 1 PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION 2 Till date, no study has reported cathepsin K levels in GCF before and after periodontal therapy, nor correlated them with the levels in serum. In this context, the present study is designed to assess the role of cathepsin K in periodontal disease progression and also to know the effect of periodontal treatment on cathepsin K concentration. 6.2 REVIEW OF LITERATURE: 1. The induction and involvement of cathepsin K in the pathologic bone changes in diffuse sclerosing osteomyelitis of mandible was evaluated by immunofluorescence staining and an association between cathepsin K and pathological intramembranous bone destruction and remodeling was demonstrated. 2. Cathepsin K was assayed in serum of patients with longstanding rheumatoid arthritis by ELISA and elevated levels and significant correlation of cathepsin K with radiological destruction were found as compared to a healthy control group. 3. The changes of cathepsin K mRNA expression were examined in parallel with histologic changes in alveolar bone during orthodontic tooth movement using RTPCR and it was concluded that site-specific early induction of cathepsin K mRNA may cause an imbalance in the relative resorption activities on pressure and tension side incident to such movement. 4. Concentration of cathepsin K in GCF of normal and periodontitis patients were investigated using ELISA and an increased concentration in patients with periodontitis was reported. 5. Using ELISA the concentration of cathepsin K in GCF around dental implants was determined and clinical parameters of peri-implantitis were found to be associated with a higher amount of cathepsin K. 6.3 OBJECTIVES OF THE STUDY: 1. To estimate the levels of cathepsin K in GCF and serum in healthy and periodontally affected individuals. 2. To find out the association between cathepsin K levels in GCF and serum in the periodontal health, disease and periodontitis affected individuals after the treatment. 3. To explore the possibility of using cathepsin K as a marker of osteoclastic activity of periodontal diseases. 3 7. MATERIALS AND METHODS: - 7.1 SOURCES OF THE DATA Those to be studied will be patients referred to the outpatient section, Department of Periodontics, Government Dental College and Research Institute, Bangalore. 7.2 METHODS OF COLLECTION OF DATA Subjects will be selected randomly and categorized into 4 groups based on gingival index (Loe & Silness), probing pocket depth (≥5mm), clinical attachment loss (≥3mm) and bleeding on probing. 80 samples (40 GCF & 40 serum) from 30 subjects divided into four groups: Group I (10 patients with healthy periodontium), Group II (10 patients with gingivitis), Group III (10 patients with chronic periodontitis), Group IV (10 patients of Group III after scaling and root planing). It will be made clear to the potential subjects that participation will be voluntary and written informed consent will be obtained from those who agree to participate. The grouping is as follows: Group I : 20 samples (10 GCF and 10 serum) from 10 patients with healthy periodontium. Group II : 20 samples (10 GCF and 10 serum) from 10 patients with gingivitis. Group III : 20 samples (10 GCF and 10 serum) from 10 patients with chronic periodontitis. Group IV : 20 samples (10 GCF and 10 serum) from 10 patients of group III after treatment. INCLUSION CRITERIA: 1. Age group 25-40 years. 2. Subjects who have not received periodontal therapy, within preceeding six months. 3. Subjects should have at least 20 natural teeth. 4 EXCLUSION CRITERIA: 1. 2. 3. 4. 5. 6. 7. Smokers Arthritis (Rheumatoid and osteoarthritis) Osteoporosis Osteolytic bone metastasis The post menopausal women Any other systemic disease which can alter the course of periodontal disease. Subjects should not be on any medication like cyclosporine A, bisphosphonates, hormone replacement therapy, steroids, calcium or vitamin D. 8. Subjects should not have received any anti-inflammatory drugs and antibiotics in the previous six months. Gingival index, probing pocket depth, bleeding on probing, clinical attachment loss will be measured after GCF collection to avoid contamination of the sample with blood. The radiographs will be done to confirm site assessment. The clinical measurements will be carried out by the same examiner, using William's graduated periodontal probe. The site showing the pocket probing depth of ≥5mm with clinical attachment loss of ≥3mm, (in case of periodontitis) will be selected for GCF sample collection. GCF collection will be done using micro capillary pipettes at initial visit in Group I, Group II, Group III and in Group IV (i.e. Group III patients 8 weeks after treatment) and samples will be stored at −70 oC till the assay procedure. Blood collection: 2 ml of blood will be collected from the antecubital fossa by venipuncture using 20-guage needle with 2 ml syringes and immediately transferred to the laboratory. Serum will be extracted from blood and stored at −70 oC till the assay procedure. Estimation of cathepsin K levels will be done by using ELISA KIT obtained from ‘Biomedica™’ Austria. 7.3 Does the study require any investigations or interventions to be conducted on patients or other humans or animals? a) GCF collection will be done by using micro capillary pipettes. b) 2 ml of blood sample will be drawn from each patient by venipuncture at the antecubital fossa. 5 7.4 Has ethical clearance been obtained from your institution in case of 7.3? Yes. 8. LIST OF REFERENCES: 1. Montonen M, Li TF, Lukinmaa PL et al. RANKL and Cathepsin K in diffuse sclerosing osteomyelitis of the mandible. J Oral Pathol Med 2006; 35: 620-625. 2. Skoumal M, Haberhauer G, Kolarz G, Hawa G, Woloszczuk W, Kingler A .Serum cathepsin K levels of patients with longstanding rheumatoid arthritis: correlation with radiological destruction. Arthritis Res Ther 2005;7: R65-R70. 3. Ohba Y, Ohba T, Terai K, Moriyama K . Expression of cathepsin K mRNA during experimental tooth movement in rat as revealed by in situ hybridization. Archives of Oral Biology 2000; 45: 63-69. 4. Mogi M, Otogoto J. Expression of cathepsin K in gingival crevicular fluid of patients with periodontitis. Archives of Oral Biology 2007; 52: 894-898. 5. Strbac GD, Monov G, Cei S, Kandler B, Watzek G, Gruber R . Cathepsin K levels in the crevicular fluid of dental implants: a pilot study. J Clin Periodontol 2006; 33: 302-308. 6