RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, BANGALORE
PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION
1 NAME OF THE CANDIDATE DEPARTMENT OF CONSERVATIVE
AND ADDRESS DENTISTRYAND ENDODONTICS,
DR.SYAMALA REDDY DENTAL
DR.SYAMALA REDDY DENTAL
2 NAME OF THE INSTITUTION COLLEGE, HOSPITAL & RESEARCH
BANGALORE – 560037.
MASTER OF DENTAL SURGERY
3 COURSE OF STUDY AND CONSERVATIVE DENTISTRY &
4 DATE OF ADMISSION TO 01 JUN – 2011
5 TITLE OF THE TOPIC
“ COMPARISON OF THE ANTIBACTERIAL EFFICIENCY OF NEEM
LEAF,TULSI LEAF EXTRACTS AND TEA TREE OIL WITH 2%
SODIUM HYPOCHLORITE AGAINST E.FAECALIS- AN IN VITRO
6 BRIEF RESUME OF INTENDED WORK:
6.1 NEED FOR THE STUDY:
The major objective in root canal treatment is to disinfect the entire root
canal system. Although cleaning, shaping, and use of antimicrobial medicaments are
effective in reducing the bacterial load some bacteria do remain behind and
multiply, causing reinfection of the canal.
Considering the ineffectiveness, potential side effects and safety concerns of
synthetic drugs, the herbal alternatives for endodontic usage might prove to be
advantageous. NaOCl, chlorhexidine and calcium hydroxide have varying degree of
antibacterial properties. But they have several undesirable characteristics such as
tissue toxicity, risk of emphysema, allergic potential, disagreeable smell and taste,
and inability to remove smear layer.
Owing to the potential side effects, safety concerns and ineffectiveness of
conventional allopathic formulations, consumption of preparations from Neem,
Tulsi and Tea tree oil has increased over the last few decades. Herbs are used in
dental and medical practice for long time. Now a days because of their high
antimicrobial, anti-inflammatory, anti-oxidant and biocompatibility properties, their
use in endodontics has become more popular.
6.2 REVIEW OF LITERATURE:
In the study1, the authors compared the antimicrobial efficacy of ginger extract and
2% sodium hypochlorite against Enterococcus Faecalis using agar diffusion method.
They concluded that ginger extract showed statistically significant activity against
In this in-vitro study2, the authors compared the antibacterial efficiency of
neem leaf extract and 2% sodium hypochlorite against E.faecalis, C.albicans and
mixed culture by using the agar diffusion test. They concluded that neem leaf extract
has a significant antimicrobial effect against E.faecalis, C.albicans and mixed
culture to use as intracanal medicament.
In this study 3, the authors evaluated the antimicrobial activity of various
concentrations of Tulsi (Ocimum sanctum) extract against Streptococcus mutans by
agar diffusion test. They concluded that Tulsi extract demonstrated an antimicrobial
activity against Streptococcus mutans. It has maximum antimicrobial potential at the
concentration of 4% level.
In this study4, the authors compared the inhibitory activity of root canal
irrigants against Candida albicans, Enterococcus faecalis and Staphylococcus aureus
by agar diffusion test and redox indicator rasazurin to check the inhibitory effect and
metabolic activities of these microorganisms. They concluded that 5.25% sodium
hypochlorite is most effective against E.faecalis as compared to EDTA and citric
In this in vitro study5 , the authors checked the effect of mango and neem
extract on four microorganisms causing dental caries: Streptococcus mutans,
Streptococcus salivavius, Streptococcus mitis, and Streptococcus sanguis: by agar
diffusion test. They concluded that neem extract produced the maximum zone of
inhibition on streptococcus mutans at 50% concentration. neem even at 5%
concentration it shows zone of inhibition for all four bacteria. Mango extract at 50%
concentration showed maximum zone of inhibition on Streptococcus mitis.
6.3 OBJECTIVES OF THE STUDY:
1 To check the antibacterial efficiency of neem leaf,tulsi leaf extract and tea
tree oil against Enterococcus.Faecalis.
2 To compare the antibacterial efficiency of neem leaf, tulsi leaf extract and
tea tree oil with 2% Sodium hypochlorite.
7 MATERIALS AND METHODS:
7.1 SOURCE OF DATA
Neem leaf extract
Tea tree oil
2% Sodium hypochlorite solution
Enterococcus faecalis stains ( ATCC 29212)
Brain heart infusion (BHI) broth powder
Brain heart infusion (BHI) agar plates
7.2 METHOD OF COLLECTION OF DATA
Neem extract,tulsi extract, tea tree oil and fresh stains of E.faecalis(ATCC
29212) shall be used.
E.faecalis cultures shall be maintained on brain heart infusion (BHI)
broth. Cultures shall grow overnight at 370C in BHI broth and bacterial growth
shall be checked by changes in turbidity at 24hrs by using the McFarland scale.
To check the antimicrobial efficacy of neem leaf extract, tulsi leaf
extract, tea tree oil and 2% NaOCl, agar diffusion method shall be performed. BHI
(brain heart infusion) agar plates shall be prepared and cultures (200μl) will be
spread on to agar plates. Inoculation should be performed by using sterile swab
brush across the media. Four round wells,4mm deep and 8mm diameter shall be
punched using sterile cork borer. Named as groups A,B,C,D.
Group A – Neem leaf extract,
Group B – Tulsi leaf extract,
Group C – Tea tree oil,
Group D – 2% Sodium hypochlorite. .
Neem leaf extract, tulsi leaf extract, tea tree oil and 2% sodium
hypochlorite (50μl) each shall be added to the respective wells and the plates will be
incubated for 24hrs at 370C in an incubator.
After incubation period, plates will be removed and zones of
inhibition will be recorded with vernier caliper. Experiment will be performed 3
times and mean of the zone of inhibition will be recorded in mm.
- The results will be tabulated and statistically analyzed by using
analysis of variance (ANOVA).
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.
7.4. Has ethical clearance been obtained from your institution in case of 7.3?
8 LIST OF REFERENCES
1. Gulve N M, Gulve N D. Comparison of antimicrobial efficacy of ginger
extract and 2% sodium hypochlorite against enterococcous faecalis using
agar diffusion method. JIDA 2010; 4 (10):347-349.
2. Bohora A, Hegde V, Kokate S. Comparison of antibacterial efficiency of
neem leaf extract and 2% sodium hypochlorite against E.faecalis, C.albicans
and mixed culture- an in vitro study. Endodont 2010; 22: 10-14.
3. Agarwal P, Nagesh L, Murlikrishnan. Evaluation of the antimicrobial
activity of various concentrations of Tulsi(Ocimum sanctum) extract against
Streptococcus mutans: an in vitro study. Indian J Dent Res2010; 21(3):357-
4. Barcelos R, Gleiser R, Silva F. Inhibitory activity of root canal irrigants
against Candida albicans, Enterococcus faecalis and Staphylococcus aureus.
Braz oral res 2010; 24(4):406-412.
5. Prashant GM, Chandu GN, Murulikrishna KS, Shafiulla. The effect
of mango and neem extract on four organisms causing dental caries:
Streptococcus mutans, Streptococcus salivavius, Streptococcus mitis, and
Streptococcus sanguis: An in vitro study. Indian J Dent Res 2007;18(4):
9 Signature of Candidate: DR. KESHAV SINGLA
10 Remarks of the guide This is a genuine study, which will be
carried out by the post graduate student
under my supervision and guidance.
11 NAME AND DESIGNATION OF
DR. UDAY KAMATH
11.1 Guide PROFESSOR & HEAD
Dept of Conservative Dentistry &
DR. HINA SHETH
11.3 Co-guide (if any)
Dept of Conservative Dentistry &
11.5 Head of Department DR.UDAY KAMATH
12 12.1 Remarks of the Chairman