effectiveness of a 2% chlorhexidine solution mixed

8/7/2019 Effectiveness of a 2% Chlorhexidine Solution Mixed

1/24

Effectiveness of a 2%

chlorhexidine solution mixedwith antifungal(clotrimazoleclotrimazole))

agent and calcium hydroxideagainst Candida albicans

Dr GaneshMurthi

M.Sc.D endo student


2/24

Introduction

The presence of Candida albicans in the

infected pulp and periradicular areas has been

demonstrated through the use of light and

electron microscopy, as well as usingculture techniques.

The incidence of C. albicans in infected root

canals has been shown to vary between 7%

and 55% and occurs in approximately 11.36%of teeth with pulp lesions (7). It has also been

reported that C. albicans can use dentin as a

nutrient source.


3/24

Historical backgroundHistorical background

Several studies have tested the effect of

intracanal irrigants and antiseptic agents on C.

albicans, including sodium hypochlorite,

chlorhexidine solution and calcium hydroxide

paste. These antiseptic agents have been

shown to be very effective against C. albicans.

Waltimo evaluated different oral Candida spp.,

including C. albicans, in the presence calcium

hydroxide in vitro. They reported that Candidaspp. were resistant to calcium hydroxide and

concluded that there is a need for

supplementary agents to effectively treat

persistent apical periodontitis.


4/24

Chlorhexidine has been suggested to be a

useful endodontic irrigant and intracanal

antiseptic due to its low toxicity and excellent

anti-bacterial and anti-fungal activities.

A combination of chlorhexidine and calcium

hydroxide was tested against bacterial species

only, and was reported to be effective againstEnterococcus faecalis. It would therefore be of

value to determine the effective


5/24

AimAim

The purpose of the current study is to examine

the In vitro susceptibility ofC. albicans to a

mixture of chlorhexidine, clotrimazole(anti

fungal) and calcium hydroxide.


6/24

Materials and methodsMaterials and methods

The anti-fungal effect of 2% chlorhexidine

solution mixed with clotrimazole,

2% chlorhexidine mixed with calcium hydroxide

is evaluate against C. albicans.

Stock cultures of clinical isolates of C. albicans

were provided by the Microbiology Laboratory

of University of The Philippines, Manila andmaintained in Sabourauds dextrose agar

plates


7/24

Materials and methodsMaterials and methodsMethods Sample size Number of colony

Control sample 20 teeth ?

2% chlorhexidinemixed with calcium

hydroxide paste

20 teeth ?

2%chlohexidine mixed

with clotrimazole

(Antifungal agent)

20 teeth ?


8/24

Experimental procedureExperimental procedure

The experiments were performed in plastic

tissue culture. Clusters containing 24 wells,

each with an inner diameter of 16 mm. One mL

of test material was placed at the bottom ofeach culture well, to which 1 mL of a Candida

suspension was added. At the same time, 1 mL

of Sabouraud infusion broth media was mixed

with 1 mL of a Candida suspension in a culture

well to serve as a positive control.


9/24


For the negative control, 1 mL of Sabouraud

infusion broth was placed in a culture well.

Eight wells were thus used per test.The culture clusters plates were then incubated

at 37oC and evaluated after 1, 24 and 72 h


10/24


At the end of the incubation period, aliquots of

0.1 mL were withdrawn from each well and

transferred to tubes containing 5 mL of fresh

Sabouraud infusion broth

1 mL of a

Candidasuspension

1 mL of a

Candida+

2% CHX +

clotrimazole

1 mL of a

Candida+2% CHX +

CaoH

Group 1 Group 2 Group 3


11/24


Growth of the fungi was observed

daily, as indicated by the presence of turbidity in

the tubes. The presence of turbidity was

determined, and the purity of the cultures was

checked by the morphology of colonies onto

blood agar plates.


12/24

Statistics testStatistics test

The results were statistically analyzed using the

KruskalWall this test is for which group is

best among the three groups

Mann-Whitney Test for compare the two group

and find which group is most significantly


13/24

GeneralGeneral objectiveobjective

Effectiveness of a 2% chlorhexidine solution

mixed with clotrimazole ( anti fungal agent )

and calcium hydroxide against Candida

albicans and evaluated the effectiveness of

each group and find out which group is most

effective in reducing the fungi in root canal and

the drug interaction between the agents and

adverse effect of each agent.


14/24

specificspecific objectiveobjective

1. Determine the number colony in thepresence of the clotrimazole and 2%

chlorhexidine.

2. Determine the number colony in the

presence of the calcium hydroxide and 2%

chlorhexidine

3. Compare the two methods.

4. Which method is most effective in reducethe number of colony.

5. Which method is least effective in reduce the

number of colony


15/24

ResultsResultsDescriptive Statistics

ROUP N Minimum Maximum MeanSt

De iation

1.00 COLONY 20 567.00 976.00 754.8500 123.04996

Valid N

(list ise

20

2.00 COLONY 20 365.00 721.00 553.2500 99.97414

Valid N(listwise)

20

3.00 COLONY 20 123.00 611.00 300.7000 111.67674

Valid N(listwise) 20


16/24

Results

Kruskal-Wallis TestHO X1 = X2 = X3

= 0.05


17/24

Results

Kruskal-Wallis Test

Sample Number of

sample

Colony

median

Mean

Rank Result

Control

group

20 754.85 48.08 Chi square

test 43.588

Asymp.sig

.000***2% CHX +

CaoH

20 553.25 31.73

2% CHX +

clotrimazol

e

20 300.70 11.70

There is highly statically significantly

different between the three groups


18/24

Mann-Whitney Test

Group Meanrank Z value Mann-Whitney

U

P value

Group 1

Group 2

30.35 -5.331 3.00 0.030**

10.65Group 1

Group 3

28.23 -4.181 45.50 0.000***

12.78

Group 2Group 3

29.15 -4.844 21.00 0.075*11.55

There is highly statically significantly

different between group 1 and group 3 the

three groups


19/24

DiscussionDiscussion

The method used to evaluate inhibition of fungal

growth in this study was the dilution tube

susceptibility test, which is an effective method

to evaluate anti-fungal as well as anti-bacterialproperties of any filling material or solution.

This method allows direct contact in the

solution betweenC. albicans

and the agent tobe tested.


20/24


The growth ofC. albicans observed in thepositive controls confirmed the efficiency of the

methodology.

Three agents, chlorhexidine, clotrimazole and

calcium hydroxide, were chosen for this study.

Chlorhexidine is a broad-spectrum antimicrobial

agent that has been reported to be an effective

antiseptic in endodontic therapy owing to itsunique ability to bind to dentin and its

substantivity in the root canal system. It is also

effective against strains resistant to calcium

hydroxide


21/24


In the present investigation, mixing clotrimazole

with chlorhexidine was found to be very

effective against C. albicans at all experimental

times tested.

The anti-bacterial property of chlorhexidine

depends on its long-lasting alkalinity and has

been reported not to be affected by the addition

of clotrimazole


22/24


Such a combination will result in better diffusion

into the dentinal tubules of prepared root canal

systems with the potential to be used as a long-

term intracanal agent and therefore completeanti-microbial, as well as anti-fungal,

effectiveness can be improved.


23/24

CConclusiononclusion

In conclusion, the results of the present study

show the additive benefits achieved by

combining Chlorhexidine with clotrimazole in

the treatment ofC. albicans


24/24

Any Question ?Any Question ?

effectiveness of a 2% chlorhexidine solution mixed

Documents