RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,

BANGALORE, KARNATAKA.

ANNEXURE – II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION

1. / NAME OF THE CANDIDATE AND ADDRESS
(IN BLOCK LETTERS) / Dr. GARIMA AGARWAL

POST GRADUATE STUDENT,

DEPARTMENT OF PERIODONTICS,
BAPUJI DENTAL COLLEGE AND HOSPITAL,
DAVANGERE – 577 004.
2. / NAME OF THE INSTITUTION / BAPUJI DENTAL COLLEGE AND HOSPITAL,
DAVANGERE – 577004.
3. / COURSE OF THE STUDY AND SUBJECT / MASTER OF DENTAL SURGERY (MDS) IN PERIODONTICS
4. / DATE OF ADMISSION TO COURSE / 13/04/2009
5. / TITLE OF THE TOPIC / EFFECT OF PROPOLIS ON ALVEOLAR BONE LOSS IN EXPERIMENTAL PERIODONTITIS IN RATS – A BIOCHEMICO-HISTOMORPHOMETRIC STUDY.
6.
7. / BRIEF RESUME OF INTENDED WORK:
6.1  Need for the study:
Propolis is a resinous product that honeybees collect from living plants and use in the construction of their hives.1 The main pharmacologically active components, flavonoids, phenolic acids, and their esters provide propolis with the ability to act as a free radical scavenger and suggest potential anti-inflammatory actions.2 Propolis has several therapeutic properties such as anticariogenic, antiulcerative, analgesic and healing activities.3
Periodontitis is a chronic inflammatory disease which involves the presence of bacterial plaque causing pocket formation and alveolar bone resorption, ultimately leading to tooth loss.
Irrigation with propolis extract as an adjuvant to periodontal treatment was beneficial as it exhibited antimicrobial activity against periodontopathogenic organisms.4 Studies have provided histologic and morphologic evidence that when propolis was administered systemically, alveolar bone loss was prevented in the rat model.1
Biochemical markers such as alkaline phosphatase and calcium are important for assessment of bone metabolism rate and expression of these is closely related to osteoblast and osteoclast differentiation.5
Till date no scientific literature is available to correlate the effects of propolis on histomorphometric changes with the biochemical markers.
Hence the aim of the present study is to correlate the effects of propolis on alveolar bone loss with calcium and alkaline phosphatase levels in experimentally induced periodontitis in wistar rats.
6.2 Review of Literature:
A group of researchers, in an experimental study evaluated the effects of propolis solution (200mg/kg) in ligature induced periodontitis in 45 male Wistar rats. The findings of this study provided morphologic and histologic evidence that propolis, when administered systemically, prevented alveolar bone loss in the rat model.1
In an experimental study, the researchers claimed that the anti-inflammatory agent propolis improves wound healing in a rodent model of experimental diabetes. This work showed that a single topical treatment with the plant-derived bee resin, propolis, clearly accelerated the epithelial closure of full-thickness cutaneous wound healing.2
A review on propolis was done to discuss its anti-inflammatory and healing properties. Histologically it was observed that propolis treatment induced better healing by reducing the inflammatory response and it was possibly due to its antioxidant capacity. The results suggested that other than flavonoids, caffeic acid phenethyl ester, naringenin and salicylic acid present in propolis attributed to its anti-inflammatory action.3
A study was conducted on twenty chronic periodontitis patients to evaluate the effect of subgingival irrigation with propolis extract by clinical and microbiological parameters. Results showed a decrease in total viable counts of anaerobic bacteria, low levels of Porphyromonas gingivalis and a decrease in the number of sites with detectable presence of yeasts. Thus subgingival irrigation with propolis extract as an adjuvant to periodontal treatment was more effective than conventional treatment alone.4
An experimental study on rats observed that the concomitant administration of alendronate counteracted the bone resorption side effects of cyclosporine A induced alveolar bone loss. Also, the biochemical markers of bone metabolism were used to evaluate the action of CyclosporinA on alveolar bone. Serum calcium, Alkaline phosphatase and Osteocalcin levels were measured and the findings were co related with the histologic and stereologic observations.5
6.3 Objectives of the Study
1)  To evaluate the effects of propolis on the periodontal tissues by histopathologic analysis.
2)  To evaluate the effects of propolis on alveolar bone loss by morphometric analysis.
3)  To correlate the effect of propolis on alveolar bone loss with serum calcium
and alkaline phosphatase levels.
MATERIALS AND METHODS:
7.1  Source of Data
The Wistar rats will be procured from the Central Animal House, Department of Pharmacology, J. J. M. Medical College, Davangere. The permission for conducting the study has been obtained from the same.
7.2  Method of Collection of Data:
Inclusion Criteria
1)  Male Wistar rats.
2)  Rats weighing 300 - 330 grams.
Exclusion Criteria
Unhealthy or diseased rats before and during the period of study will be excluded.
Study Design
A total of 45 male wistar rats will be divided into 3 groups according to the mode of treatment rendered to them for 11 days. Each group consists of 15 rats.
Group 1 (control): Non-ligature group.
Group 2 (test): Ligature induced periodontitis group.
Group 3 (test): Ligature induced periodontitis group with gastric feeding of propolis (200mg/kg/day).
Periodontitis will be induced by placement of sterile 4-0 silk ligatures submarginally on the molar teeth under general anesthesia using ketamine (40 mg/kg).
At the end of the experimental period on day 12, 2ml of blood will be drawn from the medial canthus of the rat’s eye for biochemical analysis.
Then, the rats will be sacrificed for histopathological analysis under a light microscope and morphometric analysis under a stereomicroscope.
Statistical Analysis
All the data will be subjected to statistical analysis using one way ANOVA/Kruskal-Wallis ANOVA for multiple comparisons followed by post-hoc Tukey test/Mann - Whitney test for groupwise comparison.
7.3 Does the study require any investigation or interventions to be conducted on patients or other humans or animals? If so, please describe briefly.
Yes.
Study involves use of wistar rats. Ligature will be tied around rat’s molar teeth with gastric feeding of propolis. Subsequently, rats will be sacrificed for biochemical and histomorphometric investigations.
7.4  Has ethical clearance been obtained from your institution in case of 7.3?
Yes (attached along with synopsis)
8. / LIST OF REFERENCES:
1)  Toker H, Ozan F, Ozer H, Ozdemir H, Eren K, Yeler H. A morphometric and histopathologic evaluation of the effects of propolis on alveolar bone loss in experimental periodontitis in rats. J Periodontol 2008;79(6):1089-1094.
2)  McLennan SV, Bonner J, Milne S, Lo L, Charlton A, Kurup S et al . The anti-inflammatory agent propolis improves wound healing in a rodent model of experimental diabetes. Wound Rep Reg 2008;16:706-713.
3)  Ramos AFN, Miranda JL. Propolis: A review of its anti-inflammatory and healing actions. J Venom Anim Toxins incl Trop Dis 2007;13(4):697-710.
4)  Gebara ECE, Pustiglioni AN, Lima LAPA, Mayer MPA. Propolis extract as an adjuvant to periodontal treatment. Oral Health Prev Dent 2003;1(1):29-35.
5)  Spolidorio LC, Marcantonia E Jr, Spolidorio DMP, Nassar CA, Nassar PO, Marcantonia RA et al. Alendronate therapy in cyclosporine-induced alveolar bone loss in rats. J Periodont Res 2007;42:466-473.