Croinflon

Andressa R. P. L

Supported by grants No. 2011/0132

aDoctoral student, Department of D(UNESP), Araraquara.bDoctoral student, Department of D(UNESP), Araraquara.cPostdoctoral student, Department oUniversity (UNESP), Araraquara.dDoctoral student, Department of D(UNESP), Araraquara.eUndergraduate student, DepartmenUniversity (UNESP), Araraquara.fUndergraduate student, DepartmenUniversity (UNESP), Araraquara.gFull Professor, Department of Dent(UNESP), Araraquara.hAssociate Professor, Department ofSchool, Ribeirão Preto.iAssistant Professor, Department of(UNESP), Araraquara.

Leite et al

ssover clinical trial of theuence of the use of adhesivebiofilm formation

eite, DDS,a Danny O. Mendoza-Marin, DDS,b

André G. Paleari, DDS, PhD,c Larissa S. Rodriguez, DDS, MSc,d

Andréia A. Roccia,e Vivian B. Policastro,f

Marco A. Compagnoni, DDS, PhD,g

Raphael F. de Souza, DDS, PhD,h and Ana C. Pero, DDS, PhDi

Araraquara Dental School, São Paulo State University (UNESP),Araraquara, São Paulo, Brazil; Ribeirão Preto Dental School,University of São Paulo (USP), Ribeirão Preto, São Paulo, Brazil

Statement of problem. Contrasting results have been reported regarding the influence of the use of adhesive on biofilmformation.

Purpose. The purpose of this study was to evaluate the influence of the use of adhesive on the formation of biofilm onthe internal surface of complete dentures and the palatal mucosa of denture wearers.

Material and methods. Thirty participants with well-fitting complete dentures were randomly divided according to theexperimental design: protocol 1, adhesive use during the first 15 days, followed by no use of adhesive over the next 15 days;protocol 2, no use of adhesives during the first 15 days, followed by adhesive use over the next 15 days. After each period,material from the mucosa and intaglio of the maxillary dentures was collected. Replicate aliquots were plated onto Petridishes containing selective media for Candida spp, Streptococcus mutans, and a nonselective culture medium. Colony-formingunits were expressed as log (CFUþ1)/mL. In addition, the internal surfaces of the maxillary and mandibular completedentures were stained and photographed. From the photographs, the total internal surface and the surface stained withbiofilm were quantified (software ImageTool 3.00), and the percentage of the biofilm-covered area (%) on the maxillaryand mandibular dentures was calculated and compared with 2-way ANOVA. For the nonselective culture medium, datawere compared with the paired-sample t test, and the Wilcoxon signed rank test was performed to compare the colony countsof Candida spp and Streptococcus mutans (a¼.05).

0-3 and 2010/00736-9 from FAPESP (São Paulo Research Foundation).

ental Materials and Prosthodontics, Araraquara Dental School, São Paulo State University

ental Materials and Prosthodontics, Araraquara Dental School, São Paulo State University

f Dental Materials and Prosthodontics, Araraquara Dental School, São Paulo State

ental Materials and Prosthodontics, Araraquara Dental School, São Paulo State University

t of Dental Materials and Prosthodontics, Araraquara Dental School, São Paulo State

t of Dental Materials and Prosthodontics, Araraquara Dental School, São Paulo State

al Materials and Prosthodontics, Araraquara Dental School, São Paulo State University

Dental Materials and Prosthodontics, University of São Paulo (USP), Ribeirão Preto Dental

Dental Materials and Prosthodontics, Araraquara Dental School, São Paulo State University

2 Volume - Issue -

Results. Similar colony counts were found with or without the use of adhesive for the mucosa and internal surfaces ofmaxillary dentures, irrespective of the culture medium (P>.05). The area of dentures covered with biofilm was influencedby the use of adhesive (P¼.025), regardless of the type of denture (P¼.121).

Conclusions. The use of adhesive did not alter the colony counts of microorganisms from the palatal mucosa and maxillarydentures of complete denture wearers during the 15-day period, but it did influence the area covered with biofilm on theinternal surfaces of the complete dentures. (J Prosthet Dent 2014;-:---)

The Journal of

Clinical ImplicationsAdhesive usage did not alter the oral microbiota of denture wearersduring the 15-day period. Therefore, adhesives may be recommendedas a useful adjunct for patients with complete dentures.

Denture adhesives improve theretention and stability of completedentures and can be used for well-fitting dentures.1,2 They provide confi-dence, comfort, improved masticatoryability, and improved quality of life1-8

and may also facilitate clinical pro-cedures by increasing the retention oftrial bases when jaw relation recordsare performed.3,9 Denture adhesivescan help socially active people withneurologic deficits adapt to new den-tures and can help those who haveundergone surgical modifications.10-15

If denture adhesives do affect bio-film formation, these products couldexacerbate the tissue inflammation(associated with the Candida spp andoral bacteria) and denture stomatitisin denture wearers.16 In addition,denture adhesives may play a role inhalitosis, a common problem in den-ture wearers, given that some bacteria,specifically Streptococcus oralis, Prevotellaoralis, Fusobacterium nucleatum, andStreptococcus mutans, are related tomalodor.17

In vivo studies have found that theuse of denture adhesives does notsignificantly alter the biofilm formationover a period of 14 days18,19 or a periodof 2 months of continuous use of theseproducts.20 However, Stafford andRussell21 reported that some dentureadhesives may support microbialgrowth, and Makihira et al22 suggestedthat denture adhesives tended to exhibityeast growth. Other in vitro studiesfound that some brands of denture ad-hesives inhibited the growth of Candida

Prosthetic Dentis

albicans23 and some malodor-relatedbacteria.17

Considering that a large number ofdenture wearers regularly use or haveused denture adhesives,6,24-26 factorsrelated to their use should be investi-gated. In addition, denture wearers aremostly older individuals with impairedmanual dexterity who may have diffi-culty removing adhesive residue, pos-sibly leading to biofilm accumulationon the denture bases.27 Informationis relatively scarce or controversialregarding the influence of these adhe-sives in the oral environment andregarding their cost, which may causemany patients to abandon their use.1,28

The objective of this in vivo studywas to investigate the influence of ad-hesive use on biofilm formation on theinternal surfaces of complete denturesand on the palatal mucosa in completedentures wearers. The average dailycost of the product was also estimated.The null hypothesis of this study wasthat the use of adhesives during theproposed period would have no influ-ence on biofilm formation on the in-ternal surfaces of dentures, irrespectiveof the type of denture, or on the num-ber of microorganisms on the palatalmucosa and maxillary dentures ofdenture wearers.

MATERIAL AND METHODS

Participants

A preliminary sample of 50 com-pletely edentulous individuals wearing

try

well-fitting dentures (32 women withmean age of 63.8 �9.1 years; 18 menwith mean age of 66.0 �10.6 years)was assessed for eligibility to partici-pate in this research. These patientswere previously treated with completedentures at Araraquara Dental School.The same dental technician fabricatedall the complete dentures.

The inclusion criteria for this studywere that the individuals should beadults wearing well-fitting bimaxillarycomplete dentures less than 2 yearsold; that the complete denturesshould be without rebase/reline ma-terials; that the individuals should bementally receptive; and that theyshould have the normal volume andresilience of residual edentulousridges. Residual ridge volume wasconsidered normal when the contourof a cross-sectional portion of theedentulous ridges displayed a grosslytriangular shape, with the baseranging between labial/buccal vesti-bules and the sides corresponding tothe bilateral linear projection of bothridge slopes.29,30 The exclusion criteriawere dysfunctions in the masticatorysystem, debilitating systemic diseases,diabetes mellitus, xerostomia, use ofdenture adhesives, and use of antibi-otics in the 3 months preceding thestudy. Denture adhesive users wereexcluded from the study because theirexperience in placing and removing theproduct might have influenced theresults.

After the initial evaluation, 12 in-dividuals did not meet the inclusion

Leite et al

- 2014 3

criteria (owing to diabetes mellitus[n¼7]; recent history of antibiotic use[n¼3]; physical limitations of locomo-tion [n¼1]; or cognitive or psychologi-cal problems [n¼1]). The remainingindividuals were invited to participatein this clinical trial, and 8 declined.Written consent was obtained fromthe participants before enrollment.This study was approved by the Insti-tutional Ethics Committee (ProtocolNo. 04⁄10) and registered in theensaiosclinicos.gov.br database.

Experimental design

A sample size calculation wasdone after the preliminary evaluationof the results from the first 11 parti-cipants. This analysis was performedwith Java applets for power and samplesize (http://homepage.stat.uiowa.edu/wrlenth/Power/). For this sample sizeestimation, preliminary data of thecolony counts for the nonselectiveculture medium from the mucosa

ExcludedNot using adhesive(n=1)Causes: disliked

consistency (1)

ExcludedDeclined to participate(n=3)Causes: broken leg (1);

moved to another city (1); illness of a spouse (1)

Pn=15 (567.2+/–mean a

Evaluan=

Evaluatin=11 (5

67.2+/–1age

•

•

1 Flow diagram of participants. (DA, de

Leite et al

was considered the primary outcomevariable of the study. A 1-sample t testwas performed considering the meanand standard deviation of the differ-ences between the groups (0.54 �0.83)from the first 11 participants. Thus, asample size of at least 20 participantswas necessary to detect significant dif-ference between the groups within apower of .79 at a¼.05. Consideringpossible losses and withdrawals, agroup of 30 participants was estab-lished to be randomly assigned to 1 ofthe 2 sequences of the study.

Thirty participants were enrolledand randomized to receive 2 sequencesof treatment proposed in alternatingperiods of 15 days as follows: protocol1, adhesive use during the first 15 days,followed by no use of adhesive over thenext 15 days; protocol 2, no use ofadhesive during the first 15 days, fol-lowed by use of adhesive over the next15 days. In this study, the denture ad-hesive evaluated was Ultra CoregaCream (GlaxoSmithKline Brasil Ltda).

Assessed for eligibilityn=50

Randomizedn=30

E

Protocol 2n=15 (5 men, mean69.6+/–11.1; 10 womean age: 69.2+/–

rotocol 1 men, mean age:11.6; 10 women,ge: 64.6+/–8.8)

tion 1 (with DA)14 (5 men)

Evaluation 1 (withon=14 (4 men)

on 2 (without DA) men, mean age:

.6; 6 women, mean: 68.5+/–8.5)

Evaluation 2 (witn=9 (2 men, mean

69.5+/–7.7; 7 womenage: 67.7+/–7.2

••

nture adhesive.) (Adapted from the CON

The protocol for each participant wasdetermined randomly by sex and age byusing computer generated numbers(BioEstat 5.0; Universidade Federal doPará).

A flow diagram of participants inthe research is presented in Figure 1. Itwas adapted from the CONSORTStatement.31 For protocol 1, 15 par-ticipants (5 men with mean age of67.2 �11.6 years; 10 women withmean age of 64.6 �8.8 years) receiveda denture adhesive and were showedhow to use it according to the manu-facturer’s instructions. For protocol 2,15 participants (5 men with mean ageof 69.6 �11.1 years; 10 women withmean age of 69.2 �8.4 years) con-tinued wearing their dentures until thefirst evaluation. After each period of 15days, evaluations were performed toassess the biofilm formation on the in-ternal surfaces of the dentures and onthe palatal mucosa of the participants.

The participants were instructedto clean their dentures and mucosa

ExcludedDeclined to participate(n=1)Causes: moved toanother city (1)

ExcludedNot using adhesive(n=5)Causes: nausea (1);improper use ofadhesive (4)

xcluded (n=20)Not meeting inclusion criteria (n=12)Declined to participate (n=8)

age:men,8.4)

ut DA)

h DA) age:, mean)

•

•

SORT Statement.31)

4 Volume - Issue -

3 times a day, after the main meals(breakfast, lunch, and dinner), not touse any mouthwash, and to removetheir dentures before sleeping.32 Afterthe participants had received instruc-tions and hygiene materials (dentifriceand toothbrush), they were assessed bythe investigator, who performed bio-film disclosure of the internal surfaceof the maxillary and mandibular den-tures with 1% neutral red solution(Dinâmica Química ContemporâneaLtda). The investigator brushed thedentures until the disclosed biofilmwas completely removed. This protocolwas performed again after the firstevaluation to quantify only the biofilmformed in the respective periods of15 days.

Biofilm quantification on theinternal surface of dentures

To quantify the biofilm on the in-ternal surface of the dentures, acomputerized method associated witha biofilm-disclosing agent based on aprevious study was used.33 The investi-gator asked the participants to removetheir dentures and place them in asterilized Petri dish (20�100 mm).Then, the internal surfaces of the den-tures were rinsed with running water(5 seconds) and air-dried (10 seconds).Afterward, a disclosing agent (1% neutralred) was rinsed on the internal surface,and the denture was rinsed withrunning water and dried again. Thedenture was then positioned on aclamp fixed on a stand at a standard-ized distance and at a 45-degree incli-nation, and the disclosed surfaces werephotographed with a digital camera(Cyber-shot DSC-F717; Sony).

The photographs were processedand the areas (total internal surfaceand surface stained with biofilm)quantified (software ImageTool, Win-dows, version 3.00; University of TexasHealth Science Center). The percentageof area covered by biofilm was calcu-lated by the ratio between the biofilmarea multiplied by 100 and the totalarea of the internal surface of thecomplete dentures.

The Journal of Prosthetic Dentis

Quantitative evaluation oforal microbiota

Material from the palatal mucosaand internal surface of the maxillarydenture of each participant was col-lected with a sterile swab. Collectingmaterial from the palatal mucosa wasperformed as soon as the patientarrived in the clinic and removed thedentures. After collection, each swabwas immersed in a tube containing5 mL of sterile saline and submitted tolaboratory procedures. Each tube wasshaken vigorously for 1 minute to pro-mote the suspension of the microor-ganisms. Dilutions were made fromreplicate aliquots of 100 mL of solutiontransferred to Eppendorf tubes con-taining 900 mL of sterile saline. Analiquot of 25 mL of the resulting sus-pensions was then plated onto sterilePetri dishes containing a nonselectivemedium (Mueller-Hinton agar) and se-lective media for Candida spp (Sabour-aud dextrose agar containing 5 mg/mLchloramphenicol) and Streptococcus mutans(SB20 agar). These procedures wereperformed in duplicate. The dishes werecovered and incubated at 37�C for48 hours under aerobic conditions(Mueller-Hinton agar and Sabourauddextrose agar for Candida spp) andanaerobic conditions (SB20 agar forStreptococcus mutans). After incubation,viable colonies on each Petri dishwere counted with a digital colonycounter (CP 600 Plus; Phoenix Indústriae Comércio Equipamentos CientíficosLtda). The estimated number of colony-forming units (CFU) per milliliter wascalculated by multiplying the meanof the number of colonies from the2 Petri dishes by the dilution factorand aliquot used. The microbial countdata obtained were expressed as log(CFUþ1)/mL.

Evaluation of average daily cost ofadhesive

To estimate the average daily costof adhesive, the adhesive tubes wereweighed on a precision balance beforeand after the product was used. A

try

mathematical formula was developedto calculate the mass of adhesive useddaily by each participant. Subsequently,the average daily cost of adhesive wasestimated as the sum of the daily massof adhesive used per participant multi-plied by the price paid per gram ofproduct divided by the number of par-ticipants in the study.

The primary outcome variable se-lected for this evaluation was the col-ony counts for the nonselective culturemedium from the palatal mucosa,because it was more representative ofthe possible effect of the treatment. Astatistical assessment of the primaryoutcomes was performed with thegeneralized estimating equations (GEE)method with an identity link functionto test period as a second dependentfactor and its interaction with the use ofadhesive. The latter served as an indi-cator of a possible carryover effect. Anexchangeable working correlation wasassumed, and generalized score statis-tics were used instead of Wald tests.Periods and groups were used as cate-gorical variables.

To quantify oral microbiota, datawere compared between the experi-mental condition (use of adhesive) andcontrol (no use). For the nonselectiveMueller-Hinton agar medium, datawere compared by means of the paired-sample t test, and the Wilcoxon signedrank test was performed to comparethe colony counts of Candida sppand Streptococcus mutans. Data on thebiofilm-covered area (%) on the maxil-lary and mandibular dentures with useof adhesive and without were com-pared with 2-way ANOVA. All analyseswere performed with a¼.05 by usingsoftware (PASW Statistics, version 18;IBM Corp).

RESULTS

Thirty individuals were recruited inthis study. During the follow-up, 10participants were lost from the study.

The total aerobe counts from themucosa (GEE, P¼.128), with a .95confidence interval of the differenceranging from �0.77 to 0.07 for the

Leite et al

Table I. Mean (with standard deviation) of log (CFUþ1/mL) fornonselective culture medium

Location Condition

Mean(StandardDeviation)

95% ConfidenceInterval of theDifference

PyLower Upper

Mucosa With DA 6.06 (0.63) �.7 .066 .097

Without DA 5.72 (0.69)

Denture With DA 5.17 (1.04) �.8 .2 .224

Without DA 4.86 (1.04)

DA, denture adhesive.yCalculated with t test for paired sample. No significant difference (P>.05).

Table II. Mean (with standard deviation) and median (with interquartilerange) of log (CFUþ1/mL) for Candida spp

Location Condition

Mean(StandardDeviation)

Median(Interquartile

Range) Py

Mucosa With DA 0.47 (1.51) 0.88 (1.58)

Without DA 0.50 (1.59) 4.65 (1.45) .317

Denture With DA 1.37 (2.29) 4.95 (1.45)

Without DA 0.43 (1.36) 4.50 (3.20) .223

DA, denture adhesive.yCalculated with Wilcoxon signed rank test. No significant difference (P>.05).

Table III. Mean (with standard deviation) and median (with interquartilerange) of log (CFUþ1/mL) for Streptococcus mutans

Location Condition

Mean(StandardDeviation)

Median(Interquartile

Range) Py

Mucosa With DA 0.88 (1.58) 3.30 (0.40) .465

Without DA 0.62 (1.52) 4.30 (0.60)

Denture With DA 0.85 (1.78) 4.10 (1.33) .144

Without DA 0.78 (1.62) 4.65 (0.53)

DA, denture adhesive.yCalculated with Wilcoxon signed rank test. No significant difference (P>.05).

Table IV. Mean (with standard deviation) of biofilm-covered area (%) formaxillary and mandibular dentures

Condition

Mean (Standard Deviation)

Maxillary Denture Mandibular Denture

With denture adhesive 1.25 (0.96) 1.79 (1.26)

Without denture adhesive 0.82 (0.55) 1.45 (1.28)

- 2014 5

Mueller-Hinton medium, were similarbetween use of adhesive and no use ofadhesive. The influence of test periodswas not significant (P¼.705), nor wastheir interaction with the use of adhe-sive (P¼.477). The P value of .477 in-dicates that the possible carryover effectwas not significantly different betweenthe 2 sequences of the study.

The colony count data for theMueller-Hinton medium were similarbetween use of adhesive and no use ofadhesive from the mucosa (P¼.097)and the maxillary dentures (P¼.224)(Table I). No statistically significantdifferences were noted between use ofadhesive and no use of adhesive for thecolony counts of Candida spp (Table II)from the mucosa (P¼.317) and themaxillary dentures (P¼.223) and forS mutans (Table III) from the mucosa(P¼.465) and the maxillary dentures(P¼.144).

The mean percentages (with stan-dard deviations) of biofilm-coveredarea (%) for each experimental condi-tion are presented in Table IV. The2-way ANOVA (Table V) found thatthe area covered by biofilm for themaxillary and mandibular dentures wasinfluenced by adhesive use (P¼.025),irrespective of the type of denture(P¼.121) (Fig. 2).

It was found that, on average, eachparticipant used 3.9 �0.3 g of dentureadhesive per day, equivalent to anaverage daily cost of US $1.92.

DISCUSSION

From the results of this study, thenull hypothesis was not rejected,because adhesive use did not influencethe primary outcome variable of thisstudy. These findings are in agreementwith previous studies,18,19 which foundin vivo that use of adhesives or no useduring a 2-week period did not alter thecolony counts of Candida spp in wearersof complete dentures.

Adhesive use produced a biofilm-covered area significantly higher thanthe area of biofilm measured afterthe nonuse adhesive period, irrespectiveof the type of denture (maxillary or

Leite et al

Table V. Two-way ANOVA for biofilm-covered area (%)

Source of VariationSum ofSquares df

MeanSquare F P

Treatment (with or without adhesive) 6.93 1 6.93 5.89 .02

Denture (maxillary or mandibular) 2.91 1 2.91 2.64 .12

Treatment�Denture 0.04 1 0.04 0.05 .83

Error 15.80 19 0.83

Total 25.64 22

P<.05 indicates significant difference.

A

B1.60

1.40

1.20

1.00

0.80

Bio

film

-cov

ered

are

a, m

ean

(%)

0.60

0.40

0.20

0.00Without adhesive With adhesive

2 Comparison of the mean percentages of thebiofilm-covered area (% maxillary and mandibulardentures) with and without denture adhesive. Differentuppercase letters mean statistical difference (2-wayANOVA, P<.05).

6 Volume - Issue -

mandibular). Sato et al9 stated thatdenture adhesives swell and becomeviscous and sticky in the presence ofsaliva, which may explain the difficultyin removing these products from theintaglio of the dentures in this study.

The type of denture (maxillary ormandibular) had no influence on bio-film formation in this study. Previousstudies33,34 have found that thedifferent shapes of dentures can signif-icantly affect biofilm accumulation. Thefact that all the participants receivedinstructions on and materials forcleaning their dentures and mucosaduring the trial period was probablyimportant in moderating the effect ofthe type of denture.

A 15-day trial was used in this study,because during this period it is notpossible to induce candidiasis.19 In arecent study, Ozkan et al20 evaluated

The Journal of Prosthetic Dentis

the extended use of adhesives in den-ture wearers for up to 2 months, andthe results indicated no increase inmicrobiota or adverse effects.

The influence of adhesive use onthe growth of S mutans was evaluated inthis study because recent studies haveindicated that C albicans, Staphylococcusaureus, and S mutans are consideredhabitual colonizers of the oral biofilmin complete denture wearers and are,therefore, strongly involved in the earlystages of biofilm formation. S mutans iscapable of producing an extracellularmatrix of polysaccharide, facilitatingthe adherence of other microorganismssuch as C albicans.35,36 In addition,S mutans is related to oral malodor.17

This study found no statisticallysignificant increase or decrease in thecolony counts of microorganisms withthe use of adhesive. The proliferation of

try

microorganisms could be due to theaccumulation of these products on thedenture surfaces.18 The fact that par-ticipants were instructed to clean theirdentures and remove any remainingadhesive before a new application mayhave had an influence on these findings.Another factor was that all participantswere using well-fitting dentures withappropriate retention and stability,providing a smaller space between thedenture base and mucosa, and conse-quently a thin layer of adhesive wasused.18,19

The harmful effects caused by den-ture adhesives are related to theirmisuse, and adhesives are contra-indicated for ill-fitting dentures.13,20 Insuch situations, thicker films of adhe-sive are used progressively to attempt tocompensate for inadequately fittingdenture bases.

Both dentists and patients shouldbe educated in the best use of dentureadhesives. Furthermore, the routine useof these products must be managed bymeans of periodic oral examinations.Pero et al24 observed that 47.6% ofthe respondents had used adhesivesby themselves, which suggests that adentist did not supervise the use ofthese products.

The average daily cost of the adhe-sive was estimated in the study. On av-erage, each participant used 3.9 �0.3 gof adhesive per day, which correspondsto an average daily cost of approxi-mately US$1.92. This represents con-siderable expense for many patients,considering that users are mostly on alow income.37 In Brazil, it representsapproximately 18% of the current mini-mum salary. In a recent survey of Bra-zilians,24 56.2% of patients said theyhad abandoned the use of a dentureadhesive because it was not effective,and 6.2% said that they had abandonedits use because of cost.

One limitation of this research wasthat 10 participants were lost from thestudy during the follow-up, because 4declined to continue and 6 reportednot using the adhesive during the trialperiod, owing to nausea (n¼1), dislikeof the consistency (n¼1), and improper

Leite et al

- 2014 7

use of the adhesive (n¼4). This lossmay have increased the chance of bias.The other 3 participants were lost dur-ing the study because of a broken leg(n¼1), a move to another city (n¼1),or a sick spouse (n¼1).

Other limitations of this study arethat only 1 brand of adhesive wasevaluated and that the ratio of womento men was 2:1, even though womenare more careful in their denture hy-giene habits.38 However, other in-vestigators have observed that sex didnot influence the prevalence of denture-related oral mucosal lesions or thenumber of microbes in the saliva ofcomplete denture wearers.39,40 Futurestudies should investigate whether sexcould influence biofilm formation indenture adhesive users.

In spite of these limitations, this isthe first study identified by the authorsin which a crossover design was used tocompare the intervention (adhesive us-age) and the control (nonuse). Thisexperimental design was used to elimi-nate the interparticipant response vari-ation to the same treatment and toreduce the influence of confoundingcovariates (because each participantserves as his/her own control), in-creasing the statistical efficacy.41,42

CONCLUSIONS

Based on the results obtained in thisstudy, it was concluded that adhesiveusage did not alter the colony counts ofmicroorganisms from the palatal mu-cosa and maxillary dentures but didinfluence the biofilm-covered area onthe internal surfaces of complete den-tures during the 15-day period.

REFERENCES

1. Felton D, Cooper L, Duqum I, Minsley G,Guckes A, Haug S, et al. Evidence-basedguidelines for the care and maintenance ofcomplete dentures: a publication of theAmerican College of Prosthodontists. J AmDent Assoc 2011;142:1S-20S.

2. Munoz CA, Gendreau L, Shanga G,Magnuszewski T, Fernandez P, Durocher J.A clinical study to evaluate denture adhesiveuse in well-fitting dentures. J Prosthodont2012;21:123-9.

Leite et al

3. Adisman IK. The use of denture adhesives asan aid to denture treatment. J Prosthet Dent1989;62:711-5.

4. Coates AJ. Denture adhesives: a review.Aust Prosthodont J 1995;9:27-31.

5. Grasso JE. Denture adhesives: changingattitudes. J Am Dent Assoc 1996;127:90-6.

6. Coates AJ. Usage of denture adhesives. J Dent2000;28:137-40.

7. Fujimori T, Hirano S, Hayakawa I. Effectsof a denture adhesive on masticatoryfunctions for complete denture wearerseconsideration for the condition ofdenture-bearing tissues. J Med Dent Sci2002;49:151-6.

8. Hasegawa S, Sekita T, Hayakawa I. Effect ofdenture adhesive on stability of completedentures and the masticatory function. J MedDent Sci 2003;50:239-47.

9. Sato Y, Kaiba Y, Hayakawa I. Evaluationof denture retention and ease of removalfrom oral mucosa on a new gel-type dentureadhesive. Nihon Hotetsu Shika Gakkai Zasshi2008;52:175-82.

10. Zarb GA, Hobkirk J. Prosthodontictreatment for edentulous patients:complete dentures and implant-supportedprostheses. 13th ed.St Louis: Mosby; 2012.p. 121-60.

11. Shay K. Denture adhesives: choosing the rightpowders and pastes. J Am Dent Assoc1991;122:70-6.

12. Chew CL, Boone ME, Swartz ML,Phillips RW. Denture adhesives: their effectson denture retention and stability. J Dent1985;13:152-9.

13. Grasso JE. Denture adhesives. Dent ClinNorth Am 2004;48:721-33.

14. Sumita YI, Otomaru T, Taniguchi H. Effectsof a denture adhesive in edentulous patientsafter maxillectomy. Gerodontology 2011;29:645-9.

15. Slaughter A, Katz RV, Grasso JE. Professionalattitudes toward denture adhesives: a Delphitechnique survey of academic prosthodon-tists. J Prosthet Dent 1999;82:80-9.

16. Arendorf TM, Walker DM. Denturestomatitis: a review. J Oral Rehabil 1987;14:217-27.

17. Polyzois G, Stefaniotis T, Papaparaskevas J,Donta C. Antimicrobial efficacy of dentureadhesives on some oral malodor-related mi-crobes. Odontology 2013;101:103-7.

18. Kim E, Driscoll CF, Minah GE. The effect of adenture adhesive on the colonization ofCandida species in vivo. J Prosthodont2003;12:187-91.

19. Oliveira MC, Oliveira VM, Vieira AC,Rambob I. In vivo assessment of the effect ofan adhesive for complete dentures on colo-nisation of Candida species. Gerodontology2010;27:303-7.

20. Ozkan YK, Uçankale M, Ozcan M, Uner N.Effect of denture adhesive on the micro-organisms in vivo. Gerodontology 2012;29:9-16.

21. Stafford GD, Russell C. Efficiency of dentureadhesives and their possible influence onoral microorganisms. J Dent Res 1971;50:832-6.

22. Makihira S, Nikawa H, Satonobu SV, Jin C,Hamada T. Growth of Candida species oncommercial denture adhesives in vitro.Int J Prosthodont 2001;14:48-52.

23. Sampaio-Maia B, Figueiral MH, Sousa-Rodrigues P, Fernandes MH, Scully C. Theeffect of denture adhesives on Candida albicansgrowth in vitro. Gerodontology 2011;29:348-56.

24. Pero A, Paleari A, Rodriguez L, Leite A,Roccia A, Compagnoni M. Subjectiveassessment of adhesives usage by completedenture wearers in a Brazilian population.Rev Odontol UNESP 2012;41:38-42.

25. Douglass CW, Shih A, Ostry L. Will therebe a need for complete dentures in theUnited States in 2020? J Prosthet Dent2002;87:5-8.

26. Ozcan M, Kulak Y, Arikan A, Silahtar E. Theattitude of complete denture wearers towardsdenture adhesives in Istanbul. J Oral Rehabil2004;31:131-4.

27. Padilha DM, Hugo FN, Hilgert JB, DalMoro RG. Hand function and oral hygiene inolder institutionalized Brazilians. J Am GeriatrSoc 2007;55:1333-8.

28. Bates J, Murphy W. A survey of an edentulouspopulation. Br Dent J 1968;124:116-21.

29. de Souza RF, Marra J, Pero AC, Regis RR,Compagnoni MA, Paleari AG. Maxillarycomplete denture movement duringchewing in mandibular removable partialdenture wearers. Gerodontology 2009;26:19-25.

30. Paleari AG, Marra J, Rodriguez LS, DeSouza RF, Pero AC, De AMF Jr, et al. A cross-over randomised clinical trial of eccentricocclusion in complete dentures. J OralRehabil 2012;39:615-22.

31. Altman DG, Schulz KF, Moher D, Egger M,Davidoff F, Elbourne D, et al. The revisedCONSORT statement for reporting random-ized trials: explanation and elaboration. AnnIntern Med 2001;134:663-94.

32. Compagnoni MA, Souza RF, Marra J,Pero AC, Barbosa DB. Relationshipbetween Candida and nocturnal denturewear: quantitative study. J Oral Rehabil2007;34:600-5.

33. Paranhos Hde F, Salles AE, Macedo LD,Silva-Lovato CH, Pagnano VO, Watanabe E.Complete denture biofilm after brushingwith specific denture paste, neutral soapand artificial saliva. Braz Dent J 2013;24:47-52.

34. Salles AE, Macedo LD, Fernandes RA,Silva-Lovato CH, Paranhos HF. Comparativeanalysis of biofilm levels in completeupper and lower dentures after brushingassociated with specific denture paste andneutral soap. Gerodontology 2007;24:217-23.

35. Kuroki K, Hayashi T, Sato K, Asai T,Okano M, Kominami Y, et al. Effect of self-cured acrylic resin added with an inorganicantibacterial agent on Streptococcus mutans.Dent Mater J 2010;29:277-85.

36. Thein ZM, Samaranayake YH,Samaranayake LP. Effect of oral bacteriaon growth and survival of Candida albicansbiofilms. Arch Oral Biol 2006;51:672-80.

8 Volume - Issue -

37. Nicolas E, Veyrune JL, Lassauzay C.A six-month assessment of oralhealth-related quality of life of completedenture wearers using denture adhesive:a pilot study. J Prosthodont 2010;19:443-8.

38. Baran I, Nalçaci R. Self-reported denturehygiene habits and oral tissue conditions ofcomplete denture wearers. Arch GerontolGeriatr 2009;49:237-41.

39. Ryu M, Ueda T, Saito T, Yasui M, Ishihara K,Sakurai K. Oral environmental factorsaffecting number of microbes in saliva ofcomplete denture wearers. J Oral Rehabil2010;37:194-201.

40. Mandali G, Sener ID, Turker SB, Ulgen H.Factors affecting the distribution and preva-lence of oral mucosal lesions in completedenture wearers. Gerodontology 2011;28:97-103.

The Journal of Prosthetic Dentis

41. Farias Neto A, Mestriner Junior W, CarreiroAda F. Masticatory efficiency in denturewearers with bilateral balanced occlusionand canine guidance. Braz Dent J 2010;21:165-9.

42. Jüni P, Altman DG, Egger M. Systematic re-views in health care: assessing the quality ofcontrolled clinical trials. BMJ 2001;323:42-6.

Corresponding author:Dr Ana Carolina PeroDepartment of Dental Materials andProsthodonticsAraraquara Dental School, Universidade EstadualPaulista (UNESP)Rua Humaitá 1680, 14801-903Araraquara, SPBRAZILE-mail: anacarolpero@foar.unesp.br

try

AcknowledgmentThe authors thank Mr João Monti Junior (dentaltechnician) for his assistance in the fabrication ofcomplete dentures.

Copyright ª 2014 by the Editorial Council forThe Journal of Prosthetic Dentistry.

Leite et al