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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
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PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION
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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.
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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.
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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.
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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.
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