Bell Raj Eapen, MD, DNB, MSc, Dip.
Geethanjali Bhas, MD, DNB.
Cyril Kumar T, Bsc, DMLT,
Dr. Murali Srinivasan, MDS, MBA,
Atlas Star Medical Center,
P.O. Box 112392,
Tel: 009714 - 3967401 Mob: 0097150-
Objective: Chlorhexidine Gluconate
has been considered a gold standard
in its use as a potent oral antiseptic
mouth rinse. However its use is primarily
limited to the dental professionals.
The study aims in checking the efficacy
of 0.2% chlorhexidine gluconate mouthrinse
on the culturable organisms of the
oral cavity and the oropharynx.
Methods & Materials:
Three sets of swabs were collected
from 20 volunteers. The first set
(A) was collected after a thorough
oral prophylaxis was carried out.
The subjects were then asked to refrain
from any form of oral hygiene measure
for 24 hours and a second set of swabs
(B) were collected. Following this
the subjects were made to rinse their
mouths with undiluted 0.2% chlorhexidine
gluconate solution for 60 seconds
and a third set of swab (C) was collected.
The swabs were then cultured for bacterial
colonies and the colonies were then
counted after 48 hours of incubation
and scored. The mean scores for each
set of samples were then calculated
and a Kruskal-Wallis test was used
for the statistical analysis in this
Results & Conclusion:
The mean counts were considerably
higher for B & C than A after
a period of total abstinence of oral
hygiene for 24 hours. The counts for
C were considerably lower than B and
were statistically significant (p
value =0.0004). In conclusion the
reduction in the bacterial colonies
clearly illustrates the efficacy of
chlorhexidine against oral microbes,
hence the use of this agent may be
recommended routinely as a pre-procedural
protocol prior to performing any dental
or oropharyngeal procedures and also
may be effectively prescribed as an
adjunct to other conventional therapies
for oral, oropharyngeal, & upper
respiratory tract infections.
Key Words: Chlorhexidine
Gluconate, Dental Plaque, Adjunct Therapy,
For decades now, Chlorhexidine
(CHX) has been considered a 'gold standard'
as an efficient antibacterial and therapeutic
oral rinse. The use of this is however
not global as a pre-procedural therapeutic
rinse. The most preferred form of CHX is
0.2%v/w chlorhexidine gluconate solution,
commonly used as an antiseptic mouth rinse
and recommended by the majority of dental
professionals worldwide. CHX is primarily
used in the treatment of gingivitis, inhibition
of dental plaque, as an adjunct to supportive
periodontal care, antiseptic oral rinse
following dental extractions and surgeries,
prevention of alveolar osteitis, treatment
of halitosis, and prevention of oral candidiasis.[2-6]
Few have even advocated the use of CHX for
the disinfection of root canals in endodontics.
While others have strongly recommended its
use in irradiated patients and those undergoing
chemotherapy for the treatment of head and
CHX 0.2% is the leading antiseptic
for controlling gingivitis, and plaque inhibition.
Although it is a potent antiseptic when
used and is known to considerably bring
down the microbial count; it is however,
more frequently employed routinely as a
postoperative preventive adjunct measure
rather than a routine preoperative procedure.
Its use, after speculation, is invariably
limited to the field of dentistry.
This study was targeted to check the efficacy
of 0.2% CHX on the culturable microorganisms
in the oral and oropharyngeal region, and
was aimed at justifying its use as a standard
protocol prior to any procedures being carried
out in the said regions and to further recommend
it as an adjunct therapy in treatment of
all oropharyngeal infections by all specialties
of medicine, not just in dentistry.
A group of 20 non-smoking
volunteers were picked with no age and sex
predilection. The group consisted of 10
men and 10 women with the mean age of the
group being 31.2 years. The volunteers were
thoroughly examined and found to be free
from any active oropharyngeal and dental
infections. Thorough complete oral hygiene
procedures were carried out on each of the
subjects prior to the study to ensure complete
plaque elimination. Three sets of swabs
were collected from each volunteer.
The first set was collected
immediately after the prophylactic procedure
was completed and labelled A. The subjects
were then instructed not to perform any
oral hygiene procedures for 24 hours (the
subjects were to refrain from procedures
such as brushing, flossing, rinsing etc.).
A second set was then collected after 24
hours and labelled sample B. After the collection
of B, the subjects were then made to rinse
with 10 ml. of undiluted 0.2% chlorhexidine
gluconate solution thoroughly for 60 seconds
(as recommended by manufacturer's instructions).
The third set of swabs were then collected
from the subjects and labelled C.
The swabs were collected from
predetermined sites in the oral cavity -
upper and lower molar regions, dorsum of
the tongue and the tonsilar region from
each of the subjects.
The swabs were then cultured
for bacterial colonies under standard incubatory
conditions on blood and chocolate agar,
which are the most common culture media
used for the culture of oropharyngeal bacteria.
Standardised streaking techniques were followed
using a sterile loop of known volume (1/500ml).
Estimation Of bacterial
numbers: The colonies were then
counted after 48 hours. The total numbers
of bacterial colonies were counted and multiplied
using a factor based on the volume of the
streaking loop. This procedure was standardized
using quality control measures and followed
on all the culture plates. The scoring
pattern used in this study is as follows:
Score 0 - No growth.
Score I - <103 colony forming units (CFU)
Score II - 103-104 CFU
Score III - 104-105 CFU
Score IV - >105 CFU
Experimental data reveals that a colony
count less than 10-6 is required to meet
the pharmacological definition of 'sterile.'
The results obtained were
then scored and tabulated in Table-1.
The mean scores were then calculated for
A, B & C and a Kruskal-Wallis test
was used for the statistical analysis of
The results of this study
indicate that the mean scores for group
A (mean=1.5500) was considerably lower than
B (mean=3.7500) & C (mean=2.6000). The
mean values for group C further showed a
reduction in the colony numbers than B.
On statistical analysis a
significant difference was found between
groups C & B (p value = 0.0004).
The oral cavity is a reservoir
for commensal and pathogenic micro-organisms.
A complete state of asepsis is hardly prevalent
in the oral cavity. This however does not
imply that there is an active infection
at all times, but certainly depicts the
constant presence of micro-organisms which
may result in their frequent transmission
to the different communicating regions of
the oral cavity such as the oropharnyx,
lungs, nasal cavity, eustachian canal, sinuses
etc. This may not pose a significant threat
in a state of normalcy, but, in the presence
of an existing infection the normal oral
micro flora itself may potentially super
add to an existing infection. Conversely,
an existing focal sepsis in the oral cavity
may act as a continual supply of pathogens
and pus resulting in a chronic insidious
recurrent oropharyngeal infection of some
sort, which is usually not responsive to
antibiotic therapy. Such cases usually resolve
with the elimination of the source of infection
but also can be significantly controlled
to a great extent at the onset itself with
a direct local reduction in the pathogenic
numbers through the use of local antiseptics
such as chlorhexidine.
The primary application of
CHX is predominantly restricted to the field
of dentistry, that too, as a post procedural
therapeutic rinse.9 The focus must however
shift, as the study clearly evidences its
potential antiseptic actions. It will indeed
be certainly beneficial if the prescription
of CHX mouth-rinse is added as an adjunct
to conventional therapy. 
The outcome of this simple
clinical trial clearly illustrates the substantial
reduction in the bacterial numbers of the
cultures obtained from samples with and
without of the use of CHX mouth rinse. It
only progresses to highlight further the
compelling need to maintain oral asepsis.
Therefore, from the obtained results the
authors conclude - that a significant decrease
in the bacterial count is present with the
use of CHX. Therefore the clinical relevance
of the study can substantiate the following
That it is imperative to use CHX mouth rinse
as a recommended 'pre-procedural' standard
protocol for all dental related procedures
and prior to any procedures performed in
the oral and oropharyngeal region.
And further propose that it
may be routinely prescribed by other practitioners
in various other specialties of medicine
routinely as a mouth rinse to augment their
conventional therapeutic methods for treating
oropharyngeal and other related infections,
so as to definitely ensure an infection
free and possibly a sterile oral and oropharyngeal
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