Effectiveness of a 2%

chlorhexidine solution mixed

with antifungal agent and
calcium hydroxide against
Candida albicans.
SUBMITTED TO PROF.RODRIGUEZ, SOCORROMc

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The microorganisms and their products are closely related to the
etiology of pulpal and periapical lesions. They can cause pulp
necrosis due to their persistence in the root canal system after
endodontic treatment and can induce a periapical inflammatory
reaction.
The presence of R
  
in the infected pulp and
periradicular areas has been demonstrated through the use of light
and electron microscopy, as well as using culture techniques. The
incidence of R  
in infected root canals has been shown to
vary between 7% and 55% and occurs in approximately 11.36% of
teeth with pulp lesions. It has also been reported that R  
can
use dentin as a nutrient source.

Several studies have tested the effect of intracanal irrigants and

antiseptic agents on R  

includingSodium hypochlorite,
chlorhexidine solution andCalcium hydroxide paste. These
antiseptic agents have been shown to be very effective against R 

.Wartime  evaluated different oral R
 spp.
Including R  
, in the presence calcium hydroxide 
 .
They reported that R
 spp. were resistant to calcium
hydroxide and concluded that there is a need for supplementary
agents to effectively treat persistent apical periodontitis.

Although yeasts have occasionally been reported in

Untreated cases (Sen et al. 1995), they have been more associated
with cases of failed endodontic treatment(Nair et al. 1990).MO
landeret al. (1998) also looked for Candida albicans and detected
three isolates from a total of 100 canals with chronic apical
periodontitis and two from 20 root canal samples without signs of
apical periodontitis. To what extent Candida species may be of
significance in the persistence of periapical infection is unknown.
However, it may be significant that such species are present and
may not respond to conventional cleaning and shaping procedures
in the same way as other organisms.

Different vehicles have been added to calcium hydroxide in an

attempt to enhance its antimicrobial activity. Estrela   tested the
anti-fungal effectiveness of different mixtures of calcium hydroxide
using saline, polyethylene glycol and camphorated
paramonochlorophenol.

They reported complete anti-fungal effectiveness after 1 h of

exposure, irrespective of the vehicle associated with the calcium
hydroxide paste.

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 against 
  . It would
therefore be of value to determine the effectiveness of this
combination of agents against R  
 

The purpose of the current study is to examine the


 susceptibility of R  
to a mixture of chlorhexidine,
antifungal agent and calcium hydroxide.
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There is no study about antifungal agent mixed with chlorhexidine
so in this study we are going to evaluated the action of fungal agent
in root canal and the drug interaction with chlorhexidine and
antifungal agent if there is any drug interaction occurred we are
going to evaluated how it will affect the action of both drug, root
canal fungal and bacteria and also any adverse effect.
The method used to evaluate inhibition of fungal growth in the
previous study was the dilution tube susceptibility test, which is an
effective method to evaluate anti-fungal as well as anti-bacterial
properties of any filling material or solution. This method allows
direct contact in the solution between R  
and the agent to be
tested.

The growth of R  
observed in the positive controls

confirmed the efficiency of the methodology. Two agents,
chlorhexidine 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 its unique ability to bind to dentin and its substantivity in
the root canal system. It is also effective against strains resistant to
calcium hydroxide

The purpose of the current study was to examine the


 susceptibility of R  
to a mixture of chlorhexidine,
antifungal agent and calcium hydroxide
4 c 


Effectiveness of a 2% chlorhexidine solution mixed with antifungal
agent and calcium hydroxide against Candida albicans and evaluated
the interaction of drug and effect, adverse action also

4 c  


1.c Determine the number colony presence of the antifungal
agent.
2.c Determine the number colony presence of the antifungal
and 2% chlorhexidine.
3.c Compare the two methods.
4.c Which method is most effective in reduce the number of
colony.
5.c Which method is least effective in reduce the number of
colony.

R



1. Antifungal efficacy of 5.25% sodium hypochlorite, 2%
chlorhexidine Gluconate, and 17% EDTA with and without an
antifungal agent.
2. Antifungal effects of sodium hypochlorite and chlorhexidine in
root canals.
3. Evaluation of the interaction between sodium hypochlorite and
chlorhexidine gluconate and its effect on root dentin.
4. Antifungal efficacy of 5.25% sodium hypochlorite, 2%
chlorhexidine gluconate, and 17% EDTA with and without an
antifungal agent.
5. Antifungal effects of sodium hypochlorite and chlorhexidine in
root canals.
6. Evaluation of the interaction between sodium hypochlorite and
chlorhexidine gluconate and its effect on root dentin.
7. Comparison of the antibacterial efficiency of neem leaf extract and
2% sodium hypochlorite against e. faecalis, c. albicans and mixed
culture - an in vitro study.
8. Effectiveness of a 2% chlorhexidine solution mixed with calcium
hydroxide against candida albicans effectiveness of a 2%
chlorhexidine solution mixed with calcium hydroxide against
candida albicans.
9. In vitro study of the effectiveness of intracanal irrigants on
candida albicans.
10.Antimicrobial action of calcium hydroxide, chlorhexidine and
their combination on endodontic pathogens.
11. Antimicrobial activity of sodium hypochlorite associated with
intracanal medication for 
  


  
inoculated in root canals.
12. Antimicrobial activity of varying concentrations of sodium
hypochlorite on the endodontic microorganismǯsantinomiesIsraeli,
a. naeslundii, candida albicans and enterococcus faecalis.
13.Clinical aspects related to endodontic yeast infections.
14. Comparison of the antibacterial efficiency of neem leaf extract
and 2% sodium hypochlorite against e. faecalis, c. albicans and
mixed culture - an in vitro study.
15. Effect of sodium hypochlorite and five intracanal medications on

  
in root canals
16.Effectiveness of a 2% chlorhexidine solution mixed with
calciumhydroxide against candida albicans
17.Evaluation of antifungal efficacy of 5% doxycycline
Hydrochloride, 2.5% sodium hypochlorite, 17%ethylenediamine
tetraacetic acid and 0.2% chlorhexidinegluconate against candida
albicans - an in vitro study.
18. In vitro evaluation of the antimicrobial activity of
chlorhexidineand sodium hypochlorite
19.In vitro study of the effectiveness ofintracanal irrigants on
candidaalbicans
20.Antifungal effect of zeolite-incorporated tissue conditioner
against Candida albicans growth and/or acid production.


R
 
The anti-fungal effect of 2% chlorhexidine solution,Antifungal agent
(°
 ) and 2% chlorhexidine mixed other agent is 2%
chlorhexidine solution,calcium hydroxide paste with a calcium
hydroxide was evaluated against C. albicans.
Stock cultures of clinical isolates of C. albicans were provided by the
Microbiology Laboratory of university of Philippines manila and
maintained in Sabouraudǯs dextrose agar plates.
A suspension was prepared by transferring three colonies from a
Sabouraudǯs dextrose agar plate, using a sterile 4 mm diameter
platinum loop, to a 10 mm Sabouraud infusion broth in a sterilised
10 mL screw capped test tube, followed by incubation for 1 week
at37oC. two such test tubes were prepared.


  


The experiments were performed in plastic tissue cultureclusters
containing 24 wells, each with an inner diameterof 16 mm. One mL
of test material was placed at thebottom of each culture well, to
which 1 mL of a R
 suspension was added. At the same time, 1
mL of Sabouraudinfusion broth media was mixed with 1 mL of
aR
 suspension in a culture well to serve as a positivecontrol.
For the negative control, 2 mL of Sabouraudinfusion broth was
placed in a culture well. Six wells werethus used per test. The
culture clusters plates were thenincubated at 37oC and evaluated
after 1, 24 and 72 h.
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. The tubes were then
vortexed, incubated at 37oC, and observed for 7 days. 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. The results were statistically analyzed using the
KruskalȂWallis test.

 

Solution
1.cControl solution
2.c2% chlorhexidine mixed with Antifungal agent (°
 )
3.c2% chlorhexidine mixed with calcium hydroxide paste

Now we have 3 groups of variables so total extracted teeth is 60

each group containing 20 teeth.
The colony were introduced in the each group after seven days the
colony were determined by microbiology lap in university of the
Philippines
Methods Sample size Number of colony

Control sample 20 teeth

2%chlohexidine mixed with 20 teeth

Antifungal agent
(°
 

2% chlorhexidine mixed 20 teeth

with calcium hydroxide
paste

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