Original Article

Antimicrobial Efficacy of Different Toothpastes and Mouthrinses:

An In Vitro Study
Manupati Prasanth1

ABSTRACT
Background: Anti-microbial agents have been used as a chemotherapeutic agent to improve oral
health. This in vitro study was carried out to determine antimicrobial efficacy of different toothpastes
and mouthrinses against the oral pathogens.
Methods: A total of five toothpastes and five mouthrinses were tested for their antimicrobial activity
against three oral pathogens namely, Streptococcus mutans (MTCC 890), Escherichia coli (MTCC
579) and Candida albicans (MTCC 854) by well agar diffusion assay. Statistical Analysis was per-
formed using a statistical package, SPSS windows version 15, by applying mean values using analy-
sis of variance (ANOVA) with post-hoc least square differences (LSD) method(α = 0.05).
Results: Toothpaste formulation A showed maximum zones of inhibition against the test organism,
Escherichia coli (P<0.001) compared to all other toothpastes formulations. Against Streptococcus
mutans and Candida albicans, the zones of inhibition were less in comparison to E.coli but were sig-
nificantly different at higher dilutions (1:8, 1:16 P<0.05) for toothpaste formulation A.
Mouthrinses formulation H showed maximum efficacy against the test organism, Escherichia coli
(P<0.001) compared to all other mouthrinse formulations. Against Streptococcus mutans, mouth-
rinses formulations F, G and J showed significant antimicrobial activity (P<0.05) compared to formu-
lation H and I.
Conclusion: In the present study, it has been demonstrated that triclosan containing toothpastes for-
mulations are more effective in control of oral microflora compared to non-triclosan containing syn-
thetic toothpastes. Among mouthrinses formulations, chlorhexidine was found to be more effective
than or as effective as triclosan against the organisms tested.
Keywords: Antimicrobial activity, Antimicrobial agents, Chlorhexidine gluconate, Mouthrinse,
Toothpaste, Triclosan.

Received: July 2010

Accepted: November 2010 Dent Res J 2011; 8(2): 85-94

Introduction
In India, as in other developing countries, a very leading to the demineralization of the tooth enamel.
significant proportion of dental problems are due In addition, other microflora like Escherichia
to microbial infections. Dental problems are of coli and Candida are also associated with active
three types, formation of dental plaques, dental caries lesions. C. albicans is th-e most common
caries and periodontal diseases.1 yeast isolated from the oral cavity. It is by far the
Dental caries is a localized, transmissible infec- fungal species most commonly isolated from in-
tious process that ends up in the destruction of hard fected root canals, showing resistance to intercanal
dental tissue. It results from accumulation of plaque medication.3 Poor oral hygiene is one of the rea-
on the surface of the teeth and biochemical activities sons for accumulation of these microbes and their
of complex micro-communities. Streptococcus mu- harmful activities.
tans is one of the main opportunistic pathogens of Periodontal diseases are bacterial infections that
dental caries,2 which plays a central role in ferment- affect the supporting structure of the teeth (gingival,
ing carbohydrates resulting in acid production, and cementum, periodontal membrane and alveolar

1
Assistant Professor,School of Microbiology, Mahatma Gandhi National Institute of Research and Social Action (MGNIR-
SA),Hyderabad, Andhra Pradesh, India.
Correspondence to: Manupati Prasanth, Email: [email protected]

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Prasanth Antimicrobial Efficacy of Toothpastes and Mouthrinses

bone). The endotoxins, hydrolytic enzymes and tox- Materials and Methods
ic bacterial metabolites are involved in this disease. Microorganisms
Gingivitis, an inflammatory condition of gum, is the Pure cultures of Candida albicans (MTCC 854),
most common form of periodontal disease. Serious Escherichia coli (MTCC 579) and Streptococcus
forms of periodontal disease that affect the peri- mutans (MTCC 890) were obtained from the Insti-
odontal membrane and alveolar bone may results in tute of Microbial Technology, Chandigarh, India.
tooth loss. Streptococci, spirochetes and bacteroides Cultures of Candida albicans (MTCC 854), Esche-
are found to be the possible pathogens responsible richia coli (MTCC 579) were cultured in nutrient
for the disease. broth (Hi-Media) at 37°C for 24 h while Candida
In many individuals, the customary oral hygiene albicans was cultured for 48 hours. Streptococcus
method of tooth brushing is, by itself, usually insuf- mutans (MTCC 890) was cultured in brain heart
ficient over a long period to provide a level of pla- infusion broth (Hi-Media) at 37°C for 24 h.
que control consistent with oral health. Consequent-
ly, the incorporation of chemical agents with anti- Evaluation of Dentifrices
plaque or antimicrobial activity into dental products The survey was aimed at knowing the brands of
has been proposed as a potential prophylactic me- toothpastes and mouthrinses that are mostly used. As
thod of reducing plaque-mediated disease.4 The use a result, five toothpastes and five mouthrinses were
of antimicrobial chemotherapeutic agent has been selected for assessment of their in vitro antimicrobial
proposed as a means of reducing the levels of oral activities. They were purchased from local markets in
Hyderabad, Andhra Pradesh, India. The composition
bacteria, specifically Streptococcus mutans.5
of these dentifrices is given in Tables 1 and 2. The
Recently, Triclosan, a low-toxicity, non-ionic
selected dentifrices solutions were made by mixing
phenolic derivative with a wide spectrum of antimi- the calculated amount of toothpastes (2.0 gm) in
crobial activity has been successfully incorporated measured volume (2 ml) of sterile pyrogen-free dis-
into toothpastes and mouthrinses, resulting in mod- tilled water to give 1:1 dilution; they were further
erate but distinct positive effects on both dental bio- diluted in sterile distilled water and four different
film and marginal inflammation or gingivitis.6 There dilutions of 1:2, 1:4, 1:8 and 1:16 were made. Simi-
is evidence indicating that the ingredients in the larly, each mouthrinse (2 ml) was mixed with 2 ml of
formula of triclosan-containing mouthwashes, in- sterile distilled water and serial dilutions were made
cluding vehicle and other active substances, may as above. Nutrient agar and brain heart infusion agar
influence its antimicrobial activity, and consequent- plates were prepared to assess the antimicrobial activ-
ly its clinical efficiency.7 ity of dentifrices against the pathogens. All other
Triclosan has been used for over 30 years in chemicals and reagents used were of analytical grade.
skincare products, such as soaps, deodorants, and
creams. In dentistry, it was first used in a European Antimicrobial assay
toothpaste in 1985.8 Today triclosan is the active The antimicrobial activity of different concentrations
ingredient in many oral hygiene formulations. of the dentifrices was determined by modified agar
McMurry et al.9 demonstrated in a study with well diffusion method.10,11 In this method, nutrient
Escherichia coli that the antiseptic activity of triclo- agar plates were seeded with 0.5 mL of 24 h broth
san is due to its ability to block the synthesis of fatty cultures of each isolate (brain heart infusion agar was
acids by inhibiting the enoyl-acyl carrier protein used for Streptococcus mutans strain). The plates
were allowed to dry for 1 h. A sterile 8 mm cork-
reductase enzyme.
borer was used to cut one central and five wells at
Dentifrices need to contain various antimicrobial
equidistance in each of the plates. 0.2 mL of the den-
agents in order to reduce, control and prevent differ- tifrice dilutions was introduced into each of the five
ent kinds of dental diseases. Many dentifrices claim wells while the same amount of sterile distilled water
to have antimicrobial properties but very little re- was introduced into the first well as control. The
search has been conducted to investigate these plates were incubated at 37°C for 24 h (48 h for yeast
claims. Based on this scanty information, the species). The antimicrobial activity was evaluated by
present study was designed to investigate antimi- measuring the diameter of zones of inhibition (in
crobial efficacy of different toothpastes and mouth- mm) (Figure 1). All the plates were made in tripli-
rinses by using standard agar well diffusion method. cates and the experiments repeated thrice.

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Prasanth Antimicrobial Efficacy of Toothpastes and Mouthrinses

Table 1. Ingredients of various toothpastes tested for antimicrobial potential

Toothpastes Ingredients as listed on packages

A Triclosan, Sodium monofluoro phosphate, Sorbitol and Flavor.
B Sodium monofluorophosphate, Calcium Carbonate, Sorbitol.
C Sorbitol, Water, Hydrated silica, Sodium lauryl Sulfate, PEG-32, Flavor, Cellulose gum,
Sodium fluoride, Sodium saccharin, Cl -16255, Cl –17200.
D Babhul, Jambhul, Lavang, Manjishtha, Dalchini, Bor, Vajradanti, Acrod,
Khair,Patang,Akkal kadha, Bakula, Jesthamadh, Kabab chini (Chirfal), Anant mul,Maifal,
Trifala (Amal,Harda,Behada), Ajwan, Calcium- Carbonate, Tragacanth gum, Sorbitol, Me-
thyl paraben sodium, propyl paraben sodium, Sodium Lauryl Sulfate, Sodium hydroxide,
Flavor, water.
E Dadima (punicagranatum), Sodium benzoate, Bonopol,
Tumburu (Xanthoxylum alatum), Babbula (Acacia arabica), Triphala, Vidanga (Embelia
ribes), Nirgundi (Vitex negundo), Vaikranta bhasma, Nimba (Azadirachta indica), Ajamoda
satva, Pilu (Salvadora persica), Irimeda (Acacia farnesiana), Khadira (Acacia catechu),
Bakula (Mimosops elengi), Sweetener, Saccharine

Table 2. Ingredients of various Mouthrinses tested for antimicrobial potential

Mouthrinses Ingredients as listed on packages

F Triclosan, Sodium fluoride, Ethyl Alcohol.
G Chlorhexidine Gluconate.
H Chlorhexidine Gluconate, Sodium fluoride, Zinc Chloride.
I Potassium Nitrate, Sodium Fluoride
J Triclosan, Sodium fluoride, Alcohol

a b c d

Figure 1. Zones of inhibition produced by toothpaste formulation A at 24 h against the three tested microorganisms at
five different dilutions. (a) Escherichia coli, (b) Streptococcus mutans, (c) Candida albicans, (d) Control.

Statistical Analysis
Results
Statistical Analysis was performed using a statistical
The results of this investigation showed that tooth-
package, SPSS windows version 15 by applying
paste formulation A had maximum zones of inhibi-
mean values using analysis of variance (ANOVA)
tion against the test organism, Escherichia coli
with post-hoc least square differences (LSD) me-
(p<0.001, Table 3) compared to all other toothpaste
thod.
formulations. In Streptococcus mutans and Candida

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Prasanth Antimicrobial Efficacy of Toothpastes and Mouthrinses

albicans, the zones of inhibition were less in com- higher dilutions (1:8, 1:16, p<0.05, Tables 4 and 5)
parison to E. coli but were significantly different at for toothpaste formulation A.

Table 3. Anti-microbial activity of dentifrice formulations against Escherichia coli

1:1Dilution 1:2 Dilution 1:4 Dilution 1:8 Dilution 1:16 Dilution

Mean value Mean value Mean value Mean value Mean value
± Std. Devia- ± Std. Devia- ± Std. Devia- ± Std. Devia- ± Std. Devia-
tion tion tion tion tion
* * * ** **
A 28.33 ± 3.512 25.67 ± 4.041 23.33 ± 4.263 21.33 ± 4.163 18.33 ± 4.163
0 (Zone of inhi-
bition in mm)
B 21.33 ± 1.528 18.67 ± 1.263 16.33 ± 1.528 13.67 ± 1.528 11.00 ± 1.000
(Zone of inhibi-
tion in mm)

C 21.33 ± 0.600 19.33 ± 0.600 17.33 ± 1.528 15.33 ± 1.528 12.67 ± 1.528
(Zone of inhibi-
tion in mm)
D 20.67 ± 1.200 18.33 ± 0.600 15.33 ± 1.528 13.67 ± 1.528 11.00 ± 1.732
(Zone of inhibi-
tion in mm)

E 20.33 ± 0.600 19.00 ± 1.000 16.67 ± 0.600 14.67 ± 0.600 12.33 ± 0.600
(Zone of inhibi-
tion in mm)
**
F 19.70 ± 2.082 17.40 ± 1.528 15.00 ± 1.000 13.00 ± 1.000 11.33 ± 0.577
(Zone of inhibi-
tion in mm)
**
G 19.33 ± 2.082 16.67 ± 1.528 14.00 ± 1.732 11.67 ± 1.155 10.00 ± 1.000
(Zone of inhibi-
tion in mm)
*
H 33.33 ± 3.215 27.00 ± 1.000 14.67 ± 4.726 10.00 ± 1.000 7.67 ± 1.155
(Zone of inhibi-
tion in mm)
**
I 20.67 ± .577 0.00 ± 0.000 0.00 ± 0.000 0.00 ± 0.000 0.00 ± 0.000
(Zone of inhibi-
tion in mm)
**
J 20.33 ± 1.155 17.67 ± 1.528 16.00 ± 1.000 14.00±1.000 12.33±.577
(Zone of inhibi-
tion in mm)

A-E are toothpaste samples while F-J are mouthrinses.

n=3, *P<0.001, **P<0.05

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Table 4. Anti-microbial activity of dentifrice formulations against Streptococcus mutans

1:1Dilution 1:2 Dilution 1:4 Dilution 1:8 Dilution 1:16 Dilution

Mean value Mean value Mean value Mean value Mean value
± Std. Devia- ± Std. ± Std. ± Std. ± Std. Devia-
tion Deviation Deviation Deviation tion
** **
A 22.33 ± 1.200 20.67 ± 1.528 18.67 ± 1.528 16.33 ± 1.528 14.00 ± 2.000
(Zone of inhibi-
tion in mm)
B 20.00 ± 1.732 17.67 ± 1.528 15.00 ± 1.000 12.33 ± 1.528 3.67 ± 2.0
(Zone of inhibi-
tion in mm)
C 19.33 ± 2.082 17.33 ± 1.200 15.33 ± 1.200 13.33 ± 1.200 9.00 ± 2.000
(Zone of inhibi-
tion in mm)
D 18.33 ± 1.200 17.00 ± 1.000 14.67 ± 0.600 12.00 ± 0.000 2.00 ± 1.200
(Zone of inhibi-
tion in mm)
E 17.33 ± 1.200 15.00 ± 1.000 12.67 ± 1.528 10.33 ± 1.155 .00 ± .000
(Zone of inhibi-
tion in mm)
**
F 27.67 ± 1.155 25.33 ± 1.528 23.33 ± 1.528 21.67 ± .577 19.67 ± .577
(Zone of inhibi-
tion in mm)
**
G 22.67 ± 1.155 20.33 ± .577 17.00 ± 1.000 14.33 ± 1.155 11.67 ± .577
(Zone of inhibi-
tion in mm)
H 17.00 ± 1.000 14.33 ± 1.528 13.00 ± 1.000 11.00 ± 1.000 9.00 ±1.000
(Zone of inhibi-
tion in mm)
I 0.00 ± 0.000 0.00 ± 0.000 0.00 ± 0.000 0.00 ± 0.000 0.00 ± 0.000
(Zone of inhibi-
tion in mm)
**
J 22.67 ± 1.528 21.00 ± 2.000 18.67 ± 2.517 16.67 ± 2.517 15.00 ± 2.646
(Zone of inhibi-
tion in mm)

A-E are toothpaste samples while F-J are mouthrinses

N = 3, *P < 0.001, **P < 0.05

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Prasanth Antimicrobial Efficacy of Toothpastes and Mouthrinses

Table 5. Anti-microbial activity of dentifrice formulations against Candida albicans

1:1Dilution 1:2 Dilution 1:4 Dilution 1:8 Dilution 1:16 Dilution
Mean value Mean value Mean value Mean value Mean value
± Std. Devia- ± Std. Devia- ± Std. Devia- ± Std. Devia- ± Std. Devia-
tion tion tion tion tion
**
A 28.33 ± 0.600 25.67 ± 0.600 22.67 ± 1.155 20.33 ± 2.082** 18.00 ± 1.732
(Zone of inhibi-
tion in mm)
B 26.00 ± 1.000 24.00 ± 1.000 22.00 ± 1.000 17.67 ± 1.528 14.33 ± 1.200
(Zone of inhibi-
tion in mm)
C 23.00 ± 2.000 20.33 ± 1.528 17.67 ± 1.528 15.33 ± 1.528 12.67 ± 1.200
(Zone of inhibi-
tion in mm)
D 24.33 ± 0.600 21.00 ± 2.000 17.67 ± 2.400 13.67 ± 1.528 10.33 ± 1.528
(Zone of inhibi-
tion in mm)
E 18.33 ± 1.200 16.67 ± 1.600 15.00 ± 1.000 12.67 ± 0.600 10.33 ± 0.600
(Zone of inhibi-
tion in mm)
**
F 26.67 ± .577 25.33 ± 0.577 24.00 ± 0.000 22.33 ± .577 20.00 ± 0.000
(Zone of inhibi-
tion in mm)
G 16.33 ± 0.577 14.33 ± 0.577 12.00 ± 0.000 10.00 ± 1.000 8.67 ± 0.577
(Zone of inhibi-
tion in mm)
H 14.33 ± 3.215 12.33 ± 3.215 10.67 ± 3.055 6.67 ± 5.859 0.00 ± 0.000
(Zone of inhibi-
tion in mm)
I 0.00 ± 0.000 0.00 ± 0.000 0.00 ± 0.000 0.00 ± 0.000 0.00 ± 0.000
(Zone of inhibi-
tion in mm)
*
J 22.33 ± .577 20.67 ± .577 19.33 ± 1.155 17.00 ± 1.732 15.00 ± 2.646
(Zone of inhibi-
tion in mm)
A-E are toothpaste samples while F-J are different mouthrinse formulations.
n = 3, *P < 0.001, **P < 0.05

Mouthrinse formulation H showed maximum ef- formulations F, G and J showed significant antimi-
ficacy against the test organism, Escherichia coli crobial activity (p<0.05) compared to formulations
(p<0.001, Table 3) compared to all other mouthrinse H and I (Table 4), while against Candida albicans,
formulations. However, mouthrinse formulations F, the zones of inhibition were statistically significant
G, I, J also showed significant difference (p<0.05, (p<0.05) for formulation F (Table 5). The mean val-
Table 3). Against Streptococcus mutans, mouthrinse ues ± standard deviation of zones of inhibition are

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Prasanth Antimicrobial Efficacy of Toothpastes and Mouthrinses

given for all the test organisms. Each experiment in formulations such as toothpastes and mouth-
was repeated thrice (n = 3). rinses. It has recently been suggested that triclosan
blocks lipid biosynthesis by specifically inhibiting
Discussion the enzyme enoyl-acyl carrier protein reductase
Maintenance of good oral hygiene is the key to the (ENR).9 Systematic reviews of six-month clinical
prevention of dental diseases. The primary etiologi- studies have concluded that formulations containing
cal factor for dental diseases is dental plaque. The triclosan and copolymer significantly improve pla-
formation of plaque on the tooth surface is characte- que control and periodontal health.15,16 In a previous
rized by the progression from a limited number of study, Sullivan et al.17 investigated toothpaste con-
pioneer microbial species to the complex flora of taining triclosan on resistant oral Streptococci and
mature dental plaque. This progression involves measured the in vitro sensitivity of Streptococci
initial adherence of bacteria to the salivary pellicle strains against triclosan.
and subsequent accumulation by growth and inter- Next to triclosan, fluorinated products such as
bacterial adherence. Ultimately, the tooth surface formulations B and C were found to have antimi-
gets coated with a dense, complex micro- crobial activities, although these were not statistical-
community that ends up in the destruction of hard ly significant; this may be due to the ingredients
enamel tissue.2 present in their formulations. These dentifrices con-
The activities of oral microflora being responsi- tained sodium monofluorophosphate and sodium
ble for mouth odor and most oral diseases are not in fluoride as active ingredients. Fluorides are abun-
doubt. The need to keep these oral organisms to a dantly used in many oral health products including
level consistent with oral health by antimicrobial toothpastes and mouthrinses as they help in caries
agent inclusion in dentifrice has been stressed.12 prevention.18 When formulated correctly and used
When these substances are added to oral products, as directed, fluoride toothpaste will help to safely
they kill microorganisms by disrupting their cell and effectively prevent tooth decay. It is well docu-
walls and inhibiting their enzymatic activity. They mented the ability of fluoride to inhibit or even re-
also prevent bacterial aggregation, slow multiplica- verse the initiation and progression of dental ca-
tion and release endotoxins.13 Several clinical stu- ries.19 However, if the bacterial challenge is too
dies have demonstrated the inhibitory effects of an- high, it is not possible for fluoride to overcome the
timicrobial dentifrice on oral bacteria and gingiva.14 challenge completely.20 In a previous study, Jen-
Data from the present study is in support of this kins21 stated that fluoride products such as tooth-
assertion as all the investigated dental care products paste and mouthrinse formulations have shown to
exhibited wide variations in their effectiveness reduce caries between 30 and 70% compared with
against the three test microorganisms, a feature that no fluoride therapy. A systematic review indicated
may have been largely due to their antimicrobial that a toothpaste containing triclosan/copolymer
active ingredients (Table 1 and Table 2). Among all provides a more effective level on plaque control
the investigated toothpastes, formulation A emerged and periodontal health than conventional fluoride
as the most effective, based on the mean diameter of toothpaste.15 The effectiveness of fluoride tooth-
the zone of microbial inhibition produced by the pastes are concentration dependent.22
toothpastes in agar well diffusion method, against Formulations D and E are herbal based products
all the three microorganisms tested. The exceptional and exhibited least effectiveness compared to the
ability of formulation A to retain its in vitro antimi- other test formulations. This may be due to the in-
crobial activity against all the three tested pathogens gredients present. Using natural medicines to cure
even at higher dilution of 1:16 is notable. This various diseases has become an increasing trend.
might be due to the presence of triclosan in its for- Herbal medicine has made significant contribution
mulation. This become more plausible as the utility to modern medical practice.23
and effectiveness of a 1% triclosan formulation in Though studies in animals and in vitro have
health care industry has been reviewed by Jones et shown the antimicrobial properties of several of
al.8 these herbs, there is no other way of knowing their
Triclosan [5-chloro-2-(2,4-dichlorophenoxy) phe- real clinical effects without a randomized clinical
nol] has been used for more than 30 years as a gener- trial. In the present study, the herbal formulations
al antibacterial and antifungal agent, which is found studied appeared to be equally effective as the fluo-

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Prasanth Antimicrobial Efficacy of Toothpastes and Mouthrinses

ride formulations, but not superior to them.24 The covered in the 1950s, it is still considered one of the
antimicrobial activity of the herbs is due to the pres- most effective anti-plaque agents in dentistry. Its
ence of secondary metabolites such as alkaloids, long-term use is limited by its disagreeable taste,
flavonoids, polyphenols, and lectins.25 Synergistic and propensity to stain teeth brown.38 Oral adminis-
interactions between the principal components of tration of antimicrobials for a prolonged period may
these herbs are considered to be a vital part of their alter natural microflora of the gastrointestinal tract.
efficacy. This synergistic activity, however, needs to Next to chlorhexidine, triclosan and sodium fluo-
be established. Many studies on herbal base tooth- ride products such as formulations F and J were
paste in control of plaque and gingivitis are re- found to have antimicrobial activities and these
ported.24,26 A systematic review concluded that her- were statistically significant but less effective when
bal toothpastes have rarely been shown to have sig- compared to chlorhexidine formulation. Triclosan is
nificantly greater anti-plaque activity than conven- a broad spectrum antimicrobial39 which has anti-
tional pastes.27 Our data are in accordance with the plaque activity. But, it is equally effective in reduc-
literature cited above. ing the S. mutans count but shows less effectiveness
With respect to mouthrinses, formulation H has against E. coli when compared to chlorhexidine.
shown highly significant reduction in Escherichia Many studies using triclosan as an anti plaque agent
coli and Streptococcus mutans count. This may be were carried out40 and have given good results.
due to the presence of chlorhexidine gluconate and Study carried out by Jenkins et al.41 using 0.2% tric-
sodium fluoride as major ingredients in their formu- losan reported significant reduction in total microbi-
lations; this observation adds information to the ear- al count in saliva. In the present study, triclosan
lier studies carried out by Spets-Happonen et al.28 showed a significant reduction in Candida albicans
and Hefti et al.29 and Streptococci mutans counts. Although data
Chlorhexidine gluconate is a cationic biguanide from toothpaste trails evaluating triclosan have been
with broad-spectrum antimicrobial action, whose encouraging, the data on triclosan used as a mouth-
effectiveness in decreasing the formation of dental wash is limited.
biofilm (plaque) and gingivitis have been demon- Formulation I was least effective compared to
strated in several clinical studies.30 Its mechanism of other test formulations, which may be due to the
action is that the cationic molecule binds to the ne- presence of potassium nitrate and sodium fluoride as
gatively charged cell walls of the microbes, destabi- active ingredients in the formulation. They lack an-
lizing their osmotic balance.31,32 Chlorhexidine for- timicrobial activity.42
mulations are considered to be the “gold standard” It is known that a balance exists in a person’s
antiplaque mouthrinses due to their prolonged broad oral microbial population. If this balance is lost, op-
spectrum antimicrobial activity and plaque inhibito- portunistic microorganisms can proliferate, enabling
ry potential.31,33 The high efficacy could be ex- the initiation of disease processes. Therefore, the
plained by its immediate bactericidal action during formulation identified as having the largest microbi-
the time of application followed by a prolonged bac- al inhibition zone and thus, probably the strongest
teriostatic action due to adsorption at the tooth sur- antimicrobial properties may not be necessarily su-
face.34 Studies involving rinsing with 0.2% chlor- perior to those with smaller diameter inhibition
hexidine gluconate twice daily for 60 seconds as zones. Because the formulation used in vivo is likely
supplement for normal mechanical oral hygiene to be diluted by saliva, the level to which antimi-
procedures resulted in less plaque formation and crobial properties are buffered or lost in dilution in
gingivitis than rinsing with a placebo.35 Clinical iso- vitro of interest.11
lates of gram-negative bacteria were found to be This testing method also functioned as a screen-
highly susceptible to chlorhexidine gluconate.36 ing method, and may not have been able to detect
Gehlen37 studied the influence of 0.2% chlorhex- the effects of a chemical agent that does not diffuse
idine mouth rinse on plaque re-growth. In spite of through the agar matrix. More importantly, the test
its better efficacy against the oral infections, local was conducted in vitro, so it cannot be assumed that
delivery of the drug at the intended site was not suc- the results of antimicrobial efficacy could be pro-
cessful by conventional method. Conventionally portional or transferable to the oral cavity and trans-
0.2% chlorhexidine gluconate mouthwash is used lated into clinical effectiveness. Studies have dem-
for treatment of oral infections. Despite being dis- onstrated that the bacteria in biofilm forms such as

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Prasanth Antimicrobial Efficacy of Toothpastes and Mouthrinses

plaque have decreased sensitivity to antibacterial new triclosan mouthrinse formulation. J Clin Peri-
agents. Moreover, formulations for topical antimi- odontol 2000; 27(11): 806-9.
crobial oral use, such as mouthrinses and dentifric- 8. Jones RD, Jampani HB, Newman JL, Lee AS. Triclo-
es, must be able to penetrate the biofilm matrix and san: a review of effectiveness and safety in health care
settings. Am J Infect Control 2000; 28(2): 184-96.
deliver the active agents quickly because exposure
9. McMurry LM, Oethinger M, Levy SB. Triclosan tar-
times are limited under actual conditions. Neverthe- gets lipid synthesis. Nature 1998; 394(6693): 531-2.
less, the in vitro method is a well-established tech- 10. Popoola TOS, Yangomodou OD, Akintokun AK.
nique that commonly is used in screening the anti- Antimicrobial Activity of Cassava Seed Oil on Skin
microbial efficacy of chemicals before in vivo test- Pathogenic Microorganisms. Res J Medicinal Plant
ing. 2007; 1(2): 60-4.
11. Barry AL, Thornsberry C. Susceptibility Tests: Dif-
Conclusion fusion Test Procedures. In: Balows A, editor. Ma-
Results from this study have shown that triclosan nual of Clinical Microbiology. 5th ed. Boston: Asm
containing toothpaste formulations were more effec- International; 1991: p. 1117-25.
12. Ciancio S. Improving oral health: current considera-
tive in controlling the oral microflora compared to tions. J Clin Periodontol 2003; 30 (Suppl 5): 4-6.
non-triclosan containing synthetic toothpastes. 13. Bou-Chacra NA, Gobi SS, Ohara MT, Pinto TJA.
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