Effect of Essential Oil Mouthwash on Halitosis

Abstract

Halitosis is an embarrassing symptom with a significant social impact. Periodontal disease, tongue coating, interdental food impaction, and dental cavities are the predominant causative factors. The aim of this study is to evaluate the effect of essential oil mouthwash on halitosis. 30 patients aged between 16-25 were enrolled in this single-blind, parallel-group study. Informed consent was obtained. The following parameters were recorded: gingival index, plaque index, organoleptic breath assessment, and BANA test from tongue coating samples prior to treatment with essential oil mouthwash (group 1) or a placebo (group 2) at baseline and 28 days after the start of treatment. Histological examinations were performed from gingival tissue. There were significant differences (p<0.05) in plaque and gingival index after treatment with essential oil mouthwash compared to the placebo. The results provide a statistically significantly greater level of efficacy in controlling established plaque and gingival index after use of essentia it can be recommended in halitosis therapy.

Full text

http://www.revistadechimie.ro REV.CHIM.(Bucharest)♦68♦No. 3 ♦2017
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Effect of Essential Oil Mouthwash on Halitosis
RALUCA SABAU1, ALINA ORMENISAN1, ADRIANA MONEA1, MELINDA SZEKELY1, ADINA COSARCA1, TUDOR PETRU IONESCU2,
GABRIELA BERESESCU1*, CAMELIA ELENA DALAI3
1 University of Medicine and Pharmacy Targu Mures, Faculty of Dental Medicine, 38 Gh. Marinescu, 540139, Targu Mures, Romania
2 itu Maiorescu University of Bucharest, Faculty of Dental Medicine, 67A Gh. Petrascu, 031593, Bucharest, Romania
3 University of Oradea, Faculty of Medicine and Pharmacy, 10 Piata 1 Decembrie Str., 410073, Oradea, Romania
Halitosis is an embarrassing symptom with a significant social impact. Periodontal disease, tongue coating,
interdental food impaction, and dental cavities are the predominant causative factors. The aim of this study
is to evaluate the effect of essential oil mouthwash on halitosis. 30 patients aged between 16-25 were
enrolled in this single-blind, parallel-group study. Informed consent was obtained. The following parameters
were recorded: gingival index, plaque index, organoleptic breath assessment, and BANA test from tongue
coating samples prior to treatment with essential oil mouthwash (group 1) or a placebo (group 2) at
baseline and 28 days after the start of treatment. Histological examinations were performed from gingival
tissue. There were significant differences (p<0.05) in plaque and gingival index after treatment with essential
oil mouthwash compared to the placebo. The results provide a statistically significantly greater level of
efficacy in controlling established plaque and gingival index after use of essential oil mouthwash. Therefore,
it can be recommended in halitosis therapy.
Keywords: halitosis, essential oil, histologyplaque index, organoleptic breath, BANA test
Halitosis or bad breath is an embarrasing sympoms
affecting millions of people all over the world. Each year
professionals have struggled to deal this aspects, most of
the time they came at a lost [1].
The term halitosis comes from Latine
halitus osis
meaning the pathologically modified air breath out [2]. The
symptoms as a multifactorial ethology, however the most
important cause is considered to be the decomposition of
organic residues by the oral microorganisms. The first study
on halitosis appeared in 1874 and in 1934 osmoscoup was
created that is an instrument which measures the density
of small within the oral cavity in a semi-quantitative and
subjective manner. In the 70s the main causes of halitosis
was discovered as representing the sulphuric volatile
components (VSC) and in the year to come other
instruments were devised by means of which direct
identifications of VSC was possible( hydrogen-sulphuric,
methan-ethiol and dimethyl-sulphuric).
The organoleptical method is another way of measuring
halitosis. It is a highly subjective method with very good
quality results, but very poor quantity-wise since it manly
depends on the olphactiv acuity of the examiner. This is
the main reason why objective instruments had to be
developed, at the moment halimeter and BANA test being
the most widely used [1].
Whenever halitosis is diagnosed it is necessary to be
differentiating it from false halitosis (halitofobia) or a
transitional halitosis caused by diet or dehydration. Bad
breath can also be cause by NMY or respiratory diseases,
digestive diseases such as GER or NS or hepatic cirosis.
However, 90% of the halitosis cases have the oral cavity as
starting point [3].
Halitosis can be caused by the pathology of the oral
cavity such as dental caries, cauting tongue, expose dental
pulp, food impaction, periodontal diseases, alterations or
oral cancers. The main factor in halitosis is bacterian as
Cocci and Gram-negative anaerob bacily have VSC as final
metabolic products [1].
* email: gabriela.beresescu@gmail.com; Phone: 0745134184
Halitosis is a condition with a high social impact,
eighteen million people suffering from it over to million
dollars being spent annually on products attempting to
disguise the embarrassing small [2].
The aim of this study is to extend research on the effect
of various mouthwashes on halitosis and also on periodontal
parameters, since most of the time halitosis is connected
to periodontal disease. The paper also aims at evidencing
the possibilities of using BANA test as means of diagnosis
in halitosis and of early stages of periodontal disease.
Experimental part
30 patients aged between 16-25 were enrolled in the
study. All of them had corresponding oral hygiene with more
than 10% of the site showed plaque index 1 and gingival
index 1, all of them with halitosis. None of the patients had
used antibiotics for the previous four weeks, were not
subjected to orthodontic treatment and did not showed
any systemic diseases at the moment of the investigations.
None of the patients was smoker or presenting scrotal
tongue. Informed consent was obtained after clear
explanation of the stages of the study and its aims. The 30
patients were randomly divided in 2 groups of 15 subjects:
control group and test group.
The oral cavity was assessed by the same calibrated
examiner mainly focusing on the periodontal parameters.
These parameters were measured before treatment and
28 days after the start of it. Each tooth was examined for
presence or absence of bacterian plaque and gingival
bleeding. The vestibular site of the first upper molars, lingual
site of the first lower molars and vestibular site of the left
and right upper central incisors were examined.
The presence or absence of plaque was evaluated with
plaque index (O’Leary et al.1972) [5]. Gingival
inflammation was assessed with gingival index (Loe and
Silness, 1963) [6]. Halitosis was subjectively assessed by
the subjects and objectively by the examiner both
organolepticaly and using BANA tests. Breath odour
examinations were carried out objectively and subjectively.

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The patients was instructed not to use onion, garlic, spicy
food, alcohol, or smoke 48 h prior to examination, and not
to use chewing gum, mouthwash or any other substance
that could disguise halitosis prior to examination.
Objective examination (ORGO) was carried out by the
same calibrated dental examiner so that the subjective
perceptions would not modified the values in the test. The
examiner had a normal sense of smell and did not intake
alcohol, coffee, tea or cigarettes prior to examination.
Perfumed cosmetics were avoided. The examination took
place in the dental medicine surgery and the subjects were
instructed to keep the mouth completed close for 3 min
while breathing through the nose only. After 3 min they
were intrusted to slowly release the air through the mouth
and 10 cm from the nose of the examiner who graded the
smell on a scale from 0 to 5.
Subjective examination (ORGS) was carried out by the
patients before 28 days after the treatment. Each patient
was instructed to keep the mouth completed close for 3
min, after which to lick their wrist after drying to grade the
odour on a scale from 0 to 5 [7]:
Scor 0 – no odour. Odour not detectable.
Scor 1 – slight odour. Slight detectable odour, but not
characteristic to halitosis.
Scor 2 – detectable odour. Detectable odour close to
halitosis.
Scor 3 – moderate odour. Detectable odour easily
recognisable, characteristic for halitosis.
Scor 4 – strong odour. Strong odour that can be tolerated
by the examiner.
Scor 5 – severe odour. Sever odour that cannot be
tolerated by the examiner.
BANA tests(N-benzoyl-DL-arginine naphthylamide),
describe by Loesche et al. [8], is a rapid test for evaluation
of non-sufuros compounds. BANA tests were used to
evidence the three bacterian species, Porphyromonas
gingivalis, Tannarella forsythia, Treponema denticola,
which can be felt responsible for halitosis because of their
proteolotic activity. These microorganisms release an
enzyme able to hydrolyze the synthetic peptide benzoyl-
DL-arginine-naphthylamide (BANA) on the test band. When
either or all three species are present on the sample from
the dorsal side of the tongue, BANA is hydrolyze, the test
reads positive and the index became blue. The bluer the
color, the higher the bacterian concentration. As BANA
test are sensitive to light and humidity, they will be taken
out the box only prior to use and the box be seal back. The
upper part of the band provides place for name and data
examination.
The samples from the dorsal site of the tongue was
taken on a cotton swab then place on the marked lower
part of the BANA band. After placing it, the upper part of the
test which is pink-orange is dampened with distilled water
by means of sterile swab. Care should be taken not to used
to much water since it can dilute the reactive and induce a
false negative response. The test is then folded on the
dotted area so that the side containing the reactive will
came in contact with the side with the sample from the
tongue. The folded test is next introduced in a incubator in
55C for 5 min, after which is it taken out, cut on the dotted
line and the upper part of the test, with the reactive is
examined. The results were logged in accordance with
the blue zones as per manufactures instruction. The card
contains three codes: Negative – no color changes; Slight
positive – light blue dots randomly spread on the band;
Positive – dark blue patches on light brown test band.
The biopsy cores collected were submitted for
histological examination. The histological examination
was carried out according to a specific protocol: fixation
in Lille neutral formaldehyde for 5 days, dehydration in
consecutive solutions of alcohol, immersion in xylene,
inclusion in paraffin, sectioning, staining with
Hematoxiline-Eosine and examination under optic
microscope at different magniffications.
The control mouthwash was distilled water for placebo,
and the test mouthwash had the following recipe: distilled
water, essential oil of lemon, essential oil of meant, Natrium
dicarbonate .
After examination of the subjects, registering the
organoleptic scores and the BANA test results, the patients
were examined and instructed on the correct techniques
of the brushing and the cleaning the mouth. The subjects
received the same type of tooth paste and the same amount
of mouthwash according to the group: control group –
placebo mouthwash and test group – test mouthwash.
The mouthwash was used twice daily for 28 days.
Results and discussions
Halitosis represents a condition largely spread all over
the word and numerous attempts have been made to
eliminate it. The present paper evidences the effect of
essential oil mouthwash on halitosis, plaque index,
bleeding on probing index, bacterian concentration and
organoleptic examination carried out by both examiner
and subject.
The initial results showed significant changes after
treatment, with a dramatic difference in the test group
regarding the plaque index, bleeding on probing, gingival
index, BANA test results and organoleptic examinations
values.
The plaque index showed significantly higher changes
in the test group as compared with the control group. Prior
the treatment, the control group (fig.1) had an average
plaque index in all subjects of 1.333, and after treatment it
decreased to 1.211, which represents merely 9.25%
(p<0.05). The difference could be due to the higher
attention to oral hygiene during the study. The test group
(fig.2) showed a decrease in the plaque index of 31.5%,
from 1.388 before treatment to 0.966 after the treatment
(p<0.05).
Fig. 1. Plaque index
before and after
treatment in both
group (test group
and control group)
Fig. 2. BOP before
and after treatment
in both groups (test
group and control
group)

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Bleeding on probing showed positive results after
treatment in the control group with a difference of 6.02%
compared to baseline and a difference of 34.49% compared
to the baseline in the test group. Our observations
demonstrate the adjuvant effect of antimicrobian
mouthwash with essential oils combined with correct
brushing on adequate oral health with PI and BOP of 0 or
1(fig 2).
BANA test evidenced three anaerobe bacteriae acting
on the periodontium and bad breath. In the control group
the results of the test decreased by 4.38% while in the test
group the decrease was more significant of 38.09% (fig 3).
Organoleptical scores, both from the subjects and
examiner, are subjective which explains the differences in
assessing an odour of the same intensity.
After treatment, the patients assessment of their own
halitosis changes, especially in the test group. Four patients
in the control group considered their breath is less smelly
after placebo with a difference of 9.52% before and after
treatment. In the control group, 13 of the 15 patients
reported an improvement of their breath after using the
mouthwash with the difference 36.84% (fig 4).
Halitosis is brought about by the action of bacteria on
food debris and shed epithelial cells, which turn releases
volatile sulphur compounds.
BANA test is efficient in the periodontal diagnosis. The
test detect the
red complex species
[8].
In the present study, BANA test showed significant
associations in all periodontal parameters. Bleeding on
probing, gingival and plaque index were found to be
significant with BANA. Puscasu et al.[9] observed a
statistical correlation with the severity of periodontal status
and BANA. DeBoever [1] and Bosy demonstrated that
tongue coating samples were positive for BANA test and
the tongue coating of individuals with high
Asokan et al.[10] investigated the effect of oil pulling
with sesame oil on halitosis and the microorganism
responsable for it. Their study showed equally efective like
chlorhexidine against halitosis. The finding of present study
was similar regarding the beneficial effects of essential oil
mouthwash on the clinical and microbiological
parameters of periodontal inflammation.
Other studies on antibacterian properties of the essential
oil obtained similar results showing an improvement of
26% in the test group scores as compared with the control
group scores [11].
The association of essential oil [12] with mounthwash
has shown to be effective antiplaque agent. Ross et al.
[13] showed that essential oils annihilate microorganisms
in the presence of serum.
The results of our study provides a statistically
significantly greater level of efficacy in controlling
established halitosis, plaque and gingival index after use
the essential oil mouthwash.
Ultimately, with more research in the future, it will be
possible to arrive at better diagnostic tools along with
improved treatments and treatment options.
Conclusions
The evaluation of the results confirmed the work
hypothesis that is the essential oil have a benefic clinical
effect on halitosis and periodontal parameters. The present
study contain practical implication and can be used
whenever patients ask for help with their breath bad.
The present study improve by using a larger of number
of patient and a longer a period of treatment and also by
including antibacterian substances with benefic effect on
oral halitosis.
Acknowledgement: This paper was partialy sustained by the project
contract number 912/2015 financed by S.C.OPTOMED SRL in
collaboration with University of Medicine and Pharmacy of Targu
Mures and by the project contract number 1262/2015 financed by SC
ANDSER Medica SRL in collaboration with University of Medicine and
Pharmacy of Targu Mures
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The present study evaluated the effect of essential oil
mouthwash on halitosis using de BANA test and
organoleptic examination.
Halitosis is an embarrassing symptom with a significant
social impact. Periodontal disease, tongue coating,
interdental food impaction, dental cavities are the
predominant causative factors.

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Manuscript received: 3.06.2016