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Original Article

Effectiveness of different denture cleansing

methods on removal of biofilms formed in vivo
Renu Yadav, Vikender Singh Yadav1, Shushant Garg2, Sanjeev Mittal2, Rahul Garg3
Departments of Prosthodontics including Crown and Bridge, 1Periodontics and Oral Implantology, Surendera Dental
College and Research Institute, Sriganganagar, Rajasthan, 2Department of Prosthodontics including Crown and Bridge,
M.M. College of Dental Sciences and Research, Mullana, Ambala, 3Dental Surgeon, Department of Dental Surgery,
Haryana Civil Dental Services, General Hospital, Choutala, Sirsa, Haryana, India

ABSTRACT
Background: Denture plaque has been shown to be responsible for a variety of soft tissue changes in many denture
wearers. Proper routine cleaning of tissue surface of denture is essential to maintain healthy supporting tissue.
Aims: The aim of the present study was to evaluate the effectiveness of different denture cleansing methods
i.e., mechanical, chemical and combination method, in removal of plaque from the tissue surface of the denture.
Materials and Methods: A total of 50 maxillary dentures in healthy edentulous patients were cleaned randomly with
either denture brush, denture cleansing tablet, vinegar, denture cleansing tablet followed by brushing and vinegar
followed by brushing. At the test session, every denture with 48 hours accumulated plaque on the tissue surface, was
disclosed with a two‑tone plaque disclosing agent before and after cleaning the denture and the accumulated plaque
was scored according to plaque index.
Results: The mean difference of the pre‑ and postcleansing plaque score was statistically significant (P < 0.001) for
all treatment methods. The plaque removing effectiveness was found to be significantly greater with combination
method followed by mechanical and chemical methods. Vinegar was shown to be least efficacious as a denture
cleanser.
Conclusion: Results of present study suggests that combination of brushing and alkaline peroxides should be
encouraged to ensure efficient cleansing of dentures.
Keywords: Denture brush, denture cleansing methods, denture cleansing tablet, denture plaque, vinegar

INTRODUCTION subjects.[1,3] For effective cleansing of a denture, it is

Plaque and associated microorganisms on critical for a denture cleanser to remove plaque not
the tissue surface of the dentures are significant only from the polished surfaces of the prosthesis but,
factors in the pathogenesis of denture stomatitis.[1,2] more importantly, the unpolished tissue surface.[4]
Denture cleanliness is thus of paramount importance
to prevent the oral diseases among edentulous Dentures can be cleaned mechanically,
chemically, or through a combination of these
Access this article online
methods.[5] Mechanical methods are comprised of
Quick Response Code:
Website:
brushing (associated with water, soap, toothpastes)
and ultrasonic treatment. The utilization of ultrasonic
www.craniomaxillary.com cleansers is extremely limited due to lack of both
DOI: professional and lay information of this approach and
the discouraging cost. Ultrasonic cleansing would
10.4103/2278-9588.113582
thus make sense in an institutional environment.[6]

Correspondence to:
Dr. Vikender Singh Yadav, H. No.‑ 883/5, Street No. 6A, Patel Nagar, Gurgaon, Haryana, India. E‑mail: vikenderyadav@gmail.com

22 Journal of Cranio-Maxillary Diseases / Vol 2 / Issue 1 / January 2013

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Previous data have shown that brushing is one of
the most common methods of denture cleansing.[5]
Brushing is a simple, inexpensive and effective method
when used meticulously in removing denture biofilm.
However, abrasive action could result in the wear of
the denture base and relining materials.[7,8] Another
concern is that competency of mechanical methods is
jeopardized, especially among geriatric or physically
challenged denture wearers. In such persons, efficient
chemical denture cleansers might be an important
alternative to mechanical cleansing.
Chemical methods for cleaning dentures
include soaking in a commercial[5] (peroxides,
acids, mouthwashes and enzymes) or
household  (hypochlorides, sodium chloride,
[9]
vinegar) solution. These solutions are simple to
employ and can easily reach undercuts of the
denture base. The roughness of acrylic resin’s surface
also remains unchanged, and therefore possibly
less susceptible to biofilm accumulation. However,
among soak products, enzymes have shown less
antimicrobial activity than sodium hypochlorites[10]
and peroxides,[11] despite early reports of their
potential usefulness as denture cleansers.[12]
Hypochlorites are known to damage acrylic resins
and metal alloys.[13,14] On the other hand, alkaline
peroxide solutions present good antimicrobial
activity against denture biofilm comparable with that
of sodium hypochlorite solutions.[15] This property,
in addition to absence of odor and aftertaste,
makes peroxide solutions a good choice for denture
cleansing. However, routine usage may cause
bleaching of acrylic resin and may have a harmful
effect on soft resilient denture liners. Moreover,
effectiveness of household products like vinegar for
denture cleansing has not been reported widely[9,16]
and thus needs to be elucidated.
The combination of mechanical and chemical
methods is routinely recommended for denture
cleansing.[5,6,17] Nevertheless, several trials have
shown controversial outcomes when testing those
procedures. Better results were observed when testing
exclusively either solutions[18,19] or brushing[20] or the
combination method.[9,21]
The aim of the present study was therefore
to evaluate and compare the effectiveness of
mechanical (brushing), chemical (peroxides and
Journal of Cranio-Maxillary Diseases / Vol 2 / Issue 1 / January 2013
Yadav, et al.: Denture cleansing effectiveness
vinegar) and combination methods for the removal
of denture biofilm in edentulous subjects.
MATERIALS AND METHODS
Sixty healthy edentulous denture
wearers (28 males and 32 females; age range:
41‑69 years; mean age: 55.2  ±  8.2 years)
were selected among the patients attending the
Department of Prosthodontics. Dentures were
inserted at least one year (mean; 2.64 ± 1.03 years)
prior to study and were made from heat‑polymerized
resin. Patients were informed of the nature of the
investigation; and written informed consent was
obtained prior to enrolment. The informed consent
and research protocols were approved by the
Institutional Review Board.
Subjects were consecutively enrolled in the
study when the following inclusion criteria were
fulfilled: (i) edentulous and wearing a complete
acrylic maxillary denture (ii) included in the age
range of 40‑70 years (iii) had a good fitting denture
and/or were enrolled after completion of necessary
adjustments, if required. (iv) time of denture use more
than a year.
Exclusion criteria included: (i) Presence of partial
dentures or mandibular denture only (ii) use a denture
adhesive (iii) subjects with immunocompromised
or any debilitating pre‑existing systemic and oral
pathological condition (iv) subjects with a known
allergy to materials used in this study.
Maxillary dentures of healthy edentulous subjects
selected for the study were cleaned randomly with
the following five methods by choosing a sealed
envelope (out of a bunch of 60 identical envelops):
1. Mechanical; wet brushing with 80 strokes/min
in anteroposterior direction only using a denture
brush (STIM, DentAids, New Delhi, India)
followed by rinsing with water.
2. Chemical I; soaking of denture for 30 minutes
in alkaline peroxide solution prepared by
mixing a denture cleansing tablet comprised
of sodium perborate monohydrate (Fittydent,
Group  Pharmaceuticals Ltd., Thane, India) in
300 ml of warm water (45°C)
3. Combination I; combination between methods
1 and 2
23
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Yadav, et al.: Denture cleansing effectiveness
4. Chemical II; soaking of denture for 30 minutes
in diluted vinegar prepared by mixing 150 ml
vinegar (G.D. Foods Pvt. Ltd., Rajasthan India)
in 150 ml of warm water.
5. Combination II; combination between methods
1 and 4.
Treatment procedure
On the first appointment, maxillary dentures
were cleaned ultrasonically for 30 minutes to
perform a uniform baseline. Plaque was allowed to
accumulate for 48 hours in individuals with healthy
oral mucosa during which time they refrained from
all denture hygiene procedures.
At test session, the maxillary denture with 48 hours
of accumulation of plaque was coded and rinsed
in flow controlled running water (1000 cc/min) for
1 minute to remove any loose food debris. The
tissue surface of each maxillary denture was divided
into four sections approximately equal in area by
drawing a line anteroposteriorly at the midline and
another line perpendicular to the midline at about
the distal of second premolar region. The two‑tone
plaque disclosing agent (AlphaPlac, Dental Products
of India, Mumbai, India) was then dropped on the
tissue surface of the maxillary denture for one minute
to disclose bacterial plaque. Excess dye was gently
rinsed in flow controlled running water for 1 minute.
After the plaque accumulation was disclosed,
scoring was done for each of the quadrant according
to the plaque index[17,20] before each regimen by
an investigator  (S.G.) who was masked regarding
the treatment, as follows: 0  =  No visible plaque;
1 = Light Plaque  (≤25% of the quadrant covered);
2  =  Moderate Plaque (26‑50% of the quadrant
covered); 3 = Heavy Plaque (51‑75% of the quadrant
covered); 4  =  Very Heavy Plaque (76‑100% of the
quadrant covered). Intraexaminer reproducibility
was assessed with a calibration exercise performed
at separate occasions. Calibration was accepted if
recordings were similar at >90% level. A total denture
plaque score was obtained by summing the quadrant
scores. (Maximum score  =  16). The procedure for
recording precleansing plaque score was the same
for all the groups.
After recording the precleansing plaque score,
dentures in different groups were cleaned randomly
with either of the denture cleaning method by the
24 Journal of Cranio-Maxillary Diseases / Vol 2 / Issue 1 / January 2013
same investigator (R.Y.) followed by rinsing with flow
controlled running water for 1 minute. Two‑tone dye
was then dropped on the tissue surface of the denture
and the excess dye was gently rinsed. Postcleansing
plaque scoring was accomplished as before.
Statistical analysis
Each denture was considered as a statistical unit.
Descriptive data that included Mean  ±  Standard
Deviation were calculated for plaque score
before (precleansing) and after (postcleansing)
the treatment with denture cleansers in all the
five groups. A level of significance was set at the
probability value (P) ≤ 0.05. Differences in mean
values of precleansing and postcleansing plaque
score were statistically analyzed by paired t‑test for
within‑group comparisons whereas between‑group
differences were evaluated by one‑way analysis of
variance (ANOVA). If the differences at P ≤ 0.05 level
were found, the treatment comparisons were done by
Tukey’s honestly significant difference (HSD) multiple
comparisons test.
RESULTS
All subjects accepted for the active phase of this
study had a natural plaque accumulation with a score
of 1 or more. Out of 60 subjects, four were eliminated
because they used denture adhesive, and six patients
dropped out due to circumstances unrelated to study.
Thus a total of 50 subjects (10 in each group) were
evaluated to assess the outcome of the study.
There were no significant differences in mean
values of precleansing denture plaque score between
the five treatment groups (P  =  0.976). Statistical
analysis accomplished using the paired t‑test,
revealed a highly statistically significant difference in
the mean values of pre‑ and postcleansing plaque
score [P < 0.001, Table 1].
The statistical analysis of post cleansing plaque
scores using the one‑way analysis variance, revealed
a statistically significant difference between the
different five groups [P < 0.001, Table 2].
Application of Tukey’s HSD multiple comparisons
test showed a statistically significant difference between
Mechanical I (brushing) and Chemical II (vinegar) with
P = 0.001, between Chemical II and Combination I
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Yadav, et al.: Denture cleansing effectiveness

Table 1: Pre‑and postcleansing denture plaque score (Mean±S.D.)

Method Precleansing plaque score Postcleansing plaque score Mean difference P value*
Mechanicals 15.7±3.83 8.7±2.06 7±2.31 <0.001**
Chemical I (fittydent tablets) 15.7±2.50 10.7±2.63 5.0±1.56 <0.001**
Chemical II (vinegar) 15.2±2.25 13.1±2.56 2.1±0.74 <0.001**
Combination I (fittydent+brushing) 15.9±2.60 8.0±2.16 7.9±1.52 <0.001**
Combination II (vinegar+brushing) 15.3±2.26 8.2±2.15 7.1±0.88 <0.001**
*Paired t test; **Statistically significant

Table 2: Analysis of variance (ANOVA) of denture plaque Table 3: Tukey’s HSD multiple comparisons test between
scores between groups and within groups treatment groups
Variable Source of SS df MS F P Between groups Mean Std. P value
variation value difference error
Postcleansing Between 186.920 4 46.730 8.664 <0.001* Mechanical-Chemical I −2 1.04 0.32
denture groups Mechanical-Chemical II −4.4 1.04 0.001*
plaque score Mechanical-Combination I 0.7 1.04 0.96
Within 242.700 45 5.393 Mechanical-Combination II 0.5 1.04 0.99
groups Chemical I-Chemical II −2.4 1.04 0.16
Total 429.620 49 Chemical I-Combination I 2.7 1.04 0.09
*Statistically significant Chemical I-Combination II 2.5 1.04 0.13
Chemical II-Combination I 5.1 1.04 <0.001*
(fittydent denture cleansing tablet  +  brushing) with Chemical II-Combination II 4.9 1.04 <0.001*
P < 0.001 and between Chemical II and Combination II Combination I-Combination II −0.20 1.04 1
(vinegar + brushing) [Table 3]. *Statistically significant

chemical methods employed in the study can be

DISCUSSION
The objective of the present study was to evaluate
and compare the treatment outcome of different
denture cleansing methods in terms of plaque
removal from the tissue surface of the denture.
Among the three methods of cleaning the
denture, chemical method was shown to be least
efficacious for biofilm removal. Alkaline peroxide
solution has demonstrated better cleansing effects
than vinegar. These findings are in agreement with
those reported by Buergers et al.[22] On the contrary,
da Silva et al.[23] demonstrated that vinegar was more
effective than alkaline peroxide tablets against the
analyzed microbial species. But it should be noted
that these studies assessed in vitro outcomes of
the denture cleansing agents against the specific
microbial species rather than their effectiveness for
plaque removal. Moreover, perusal of the available
literature revealed the present clinical trial as the first
in vivo study to compare the outcomes of vinegar
and peroxide cleansers for denture plaque removal.
It can be hypothesized that variations in the
effectiveness for denture plaque removal by two
due to their different mechanism of action. Fittydent
tablet has sodium perborate as an active gradient.
A saturated aqueous solution of sodium perborate
gives, in effect, a solution of H2O2 buffered to a pH
of about 10. The H2O2 oxidizes to release oxygen.
The evolved O2, related to the observed effervescing
action of the cleanser solutions, is also supposed to
exert a mechanical cleansing effect.[24] In contrast,
vinegar is 6‑13% acetic acid which is a weak acid and
is only partially dissociated in aqueous solution. The
acid causes a slight decrease in the pH of the solution.
The amount of alkalinity in a cleanser solution above
a pH of 8.5 is considered to be the amount available
for cleaning purposes and thus is used for evaluating
the effectiveness of denture cleanser solutions.[25]
Moreover, the effervescence action present in the
fittydent tablet is absent in vinegar.
Combination method was found to be the
most efficacious denture cleansing protocol. This
is in agreement with the previous clinical trials
which reported that combination of brushing and
chemical methods was the most effective type of
denture hygiene methods.[18,21,26‑28] In the present

Journal of Cranio-Maxillary Diseases / Vol 2 / Issue 1 / January 2013 25

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Yadav, et al.: Denture cleansing effectiveness
study, combination of brushing with fittydent tablet
was relatively more effective than with the vinegar
soaking. This can be justified by the effervescence
action and greater degree of alkalinity in aqueous
solution for fittydent tablets in contrast to vinegar, as
previously mentioned.
Mechanical method i.e., brushing alone provided
intermediate outcomes. It was better than chemical
but less effective than combination method. Our
findings are in good conformity with those of previous
studies which pointed out that brushing has a better
effect regarding the denture hygiene than immersion
type cleansers.[20,27‑29] However, some clinical studies
found that brushing alone is less effective than
soaking for denture plaque removal.[18,19,30] It may be
speculated that nature and design of brush, degree
of irregularities on tissue surface of denture, manual
dexterity, study designs, etc., might have influenced
the results.
The analysis used in this study was based on the
internal surface of upper complete denture.[17,20,31,32]
This surface has greater potential for the collection
of pathogenic microorganisms and is associated
with denture stomatitis. External surfaces were not
assessed because they accumulate lesser amounts of
biofilm.[17,31,32]
The present protocol comprised soaking dentures
for 30  minutes according to manufacturer’s
recommendations. It was reported that alkaline
peroxides solutions are effective in few minutes
of soaking time.[6] This is probably due to their
mechanism of action. The alkaline peroxides
work through an oxygen‑liberating process.[24]
The liberated free radicals and oxygen, related to
the observed effervescence, are dissolved in water
and reach high concentrations immediately. On
the contrary, other researchers have reported that
chemical cleansers seem to be more effective on
new plaque and stains when the denture is soaked
for several hours or overnight due to high oxygen
concentration achieved after several hours.[33]
Further studies are needed to approach the outcome
of different immersion times.
In lieu of the present findings, it can be assumed
that an immersion type cleanser causes loosening but
not complete removal of the denture plaque. Also the
26 Journal of Cranio-Maxillary Diseases / Vol 2 / Issue 1 / January 2013
effectiveness of brushing alone seems to be limited,
probably because of the existence of irregularities in
the acrylic resins which makes difficult the mechanical
removal of microorganisms.[34] Therefore, in order to
obtain high quality denture cleaning, brushing and
immersion in chemical agents must be associated so
that plaque loosened by chemical cleansers can be
easily removed due to physical disruption of brushing.
Clinical findings of this study were not validated by
microbiological examination of the denture biofilm.
Therefore, inferences regarding reduction of specific
microbial species by different denture cleansing
methods could not be made. However, to the best of
our knowledge, this is the first in vivo study to compare
clinical outcomes of three methods using vinegar as
one of the chemical denture cleanser. Further studies
with a larger sample size and long‑term observations
would verify the findings presented here.
CONCLUSIONS
Combination was the most efficient method for
denture cleansing followed by mechanical and then
chemical. Vinegar alone was least effective as a
denture cleanser but in combination with brushing,
its results were comparable with other denture
cleansers. Therefore, present study suggests that
combination of brushing and alkaline peroxides
should be encouraged to ensure efficient cleansing
of dentures.
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How to cite this article: Yadav R, Yadav VS, Garg S, Mittal S, Garg R.
Effectiveness of different denture cleansing methods on removal of biofilms
formed in vivo. J Cranio Max Dis 2013;2:22-7.
Source of Support: Nil. Conflict of Interest: None declared.
Submission: October 07, 2012, Acceptance: December 11, 2012
27