Candida albicans biofilm formation on soft denture liners and efficacy of cleaning protocols

doi: 10.1111/j.1741‐2358.2011.00485.x
Candida albicans biofilm formation on soft denture liners and efficacy of cleaning protocols Objective: The aim of this study was to investigate Candida albicans biofilm formation on denture liners and to analyse the efficacy of cleaning protocols. Material and methods: Specimens were prepared from four silicone‐based soft denture liners. After artificial ageing and surface free energy determination, specimens were incubated with saliva (2 h) and Candida albicans ATCC 10231 for either short‐ (2.5 h) or long‐term (24 h) biofilm formation. Adherent cells were determined either after incubation of specimens with Candida albicans or after treatment with different denture cleaning protocols. Statistical analysis was performed using one‐way anova and the Games–Howell test (α = 0.05). Results: For both short‐ and long‐term biofilm formation, similar amounts of Candida albicans cells were found on the surface of the different liners (p = 0.295 and 0.178, respectively). For both short‐ and long‐term biofilm formation, the highest cleaning efficacy was observed for sodium hypochlorite (NaOCl; p < 0.01). The efficacy of the chemical denture cleaner in removing long‐term Candida albicans biofilms was significantly lower than the efficacy of removal by brushing (p < 0.001). Conclusion: Different silicone‐based soft denture liners yield similar Candida albicans biofilm formation on their surface. The highest efficacy for the removal of Candida albicans biofilms was identified for NaOCl. Chemical denture cleaners appear to have rather low efficacy to remove mature Candida albicans biofilms.     

Oral health related quality of life of edentulous patients after denture relining with a silicone-based soft liner

doi: 10.1111/j.1741‐2358.2011.00503.x Oral health related quality of life of edentulous patients after denture relining with a silicone‐based soft liner Background: Knowledge of benefits caused by a treatment on quality of life is very relevant. Despite the wide use and acceptance of soft denture liners, it is necessary to evaluate the patient’s response about the use of these materials with regard to improvement in oral health related quality of life (OHRQoL). Objectives: The aim of this study was to evaluate the influence of denture relining in the OHRQoL of edentulous patients. Materials and methods: Thirty‐two complete denture wearers had their lower dentures relined with a silicone‐based material (Mucopren soft, Kettenbach, Germany) according to chairside procedures. OHRQoL was assessed before and after 3 months of relining by means of OHIP‐EDENT, and the median scores were compared by Wilcoxon test (p ≤ 0.05). Results: After 3 months of relining, participants reported significant improvement of their OHRQoL (p ≤ 0.01). Conclusion: Denture relining with a soft liner may have a positive impact on the perceived oral health of edentulous patients.     

Effect of a denture cleanser on the concentration of volatile sulphur compounds and denture biofilm in institutionalised elderly

doi:10.1111/j.1741‐2358.2009.00341.x
Effect of a denture cleanser on the concentration of volatile sulphur compounds and denture biofilm in institutionalised elderly Objective: To determine the effectiveness of a denture cleanser in reducing the concentration of volatile sulphur compounds (VSC) and its antimicrobial action. Background: Micro‐organisms from the denture biofilm can cause local and systemic disease and halitosis. Denture cleansers are important adjuncts in oral care, but there is limited investigation on their effect in malodour compounds. Material and methods: Nineteen institutionalised elderly who wore at least an upper denture were selected; their VSC concentrations were measured and the denture biofilm was collected. In phase 1, the subjects wore their old denture and data were collected before (B₀) and after 7(A₁), 14(A₂), 28(A₃) days of continuous daily use of the denture cleanser. In phase 2, new dentures were inserted and measurements were made at 30(A_1.1), 60(A_2.2), 90(A_3.3) days of treatment. Results: The VSC concentration increased from B₀ to A₁ (p < 0.05)_, but no differences were found for the others intervals of times. Total micro‐organism data did not show a statistical difference between times in Phase I, but in Phase II, there was a statistical difference (p < 0.05) and a progressive re‐colonisation was observed. Conclusion: Within the limits of this study, it was concluded that the denture cleanser had no antimicrobial effect and VSC levels were not reduced.     

Biofilm formation of Candida albicans on the surface of a soft denture‐lining material

Background: Soft denture lining‐materials are more susceptible to microbial adhesion than hard denture base acrylic resin. Poor oral hygiene and Candida albicans infection are common among elderly denture wearers as these patients usually have difficulty in keeping them clean. Purpose: To evaluate the influence of the oral hygiene methods on the formation of a biofilm over a soft denture‐lining material. Material and methods: Twenty volunteers were randomly separated into two groups: G1 and G2. Ten volunteers performed daily hygiene of the prostheses with a soft toothbrush and toothpaste. The G2 performed a treatment identical to G1 but also immersed the prostheses in sodium hypochlorite 0.5% for 20 min, once a week. Quantification of the mean score values of biofilm formation at different times were statistically analysed using analysis of variance and Tukey’s test (α = 0.05). Results: G1 (0.65 ± 0.52) showed the lowest mean score values of biofilm formation. There was statistical difference between G1 and G2. The highest mean score values were found at 6 weeks (1.3 ± 1.08) and were statistically different from other times. Conclusion: The oral hygiene methods had a significant effect in the formation of the biofilm over a soft denture‐lining material.     

In vitro and clinical evaluation of specific dentifrices for complete denture hygiene

Objectives: To study the physical properties of two experimental dentifrices for complete denture hygiene, their effect on denture biofilm removal and antimicrobial properties by means of a clinical trial. Materials and methods: The experimental dentifrices comprised two compositions. One was based on the addition of 1% chloramine T (D1) and the other on the presence of 0.01% fluorosurfactant (D2). Measurements of density, pH, consistency, rheological features and abrasiveness were conducted. Sixty complete denture wearers were randomly assigned to three groups and were instructed to brush their dentures with a specific toothbrush: (1) Water (control); (2) D1; or (3) D2. Each method was used for 21 days. Denture biofilm was disclosed by a 1% neutral red solution and quantified by means of digital photos taken from the internal surface. Microbiological assessment was conducted to quantify Candida sp. and mutans streptococci. Data were evaluated by one‐way anova and Tukey HSD, or Kruskal–Wallis (α = 0.05). Results: Both dentifrices decreased biofilm coverage when compared with the control group. D1 was the most efficacious treatment to reduce mutans streptococci, whereas D2 showed an intermediate outcome (anova , p < 0.040). No treatment influenced Candida albicans or non‐albicans species (Kruskal–Wallis, p = 0.163 and 0.746, respectively). Conclusion: It can be concluded that brushing complete dentures with the experimental dentifrices tested could be effective for the removal of denture biofilm.     

Comparative analysis of biofilm levels in complete upper and lower dentures after brushing associated with specific denture paste and neutral soap

Objectives: The objective of this study was to compare and correlate biofilm levels in complete upper and lower prosthesis after brushing, associated with specific paste and soap, by means of computerised methodology. Materials and methods: Forty‐five complete denture wearers were selected and instructed to brush their prostheses (Soft Oral B 40) three times a day for 3 weeks with water (Control), specific paste for complete dentures (Corega Brite) (Experiment 1) and neutral soap (Experiment 2). The study was based on a cross‐over model and a wash‐out period was not included. For biofilm quantification, the internal surfaces were dyed (neutral red 1%), photographed (Canon EOS Digital) and the disclosed biofilm was measured with the Image Tool 2.0 software. The products were assessed by means of a questionnaire regarding their hygiene properties and acceptance. Results: The variance analysis indicated that the lower prostheses exhibited a mean biofilm percentage, significantly higher than the upper prostheses and that brushing with paste (Experiment 1) was more effective than soap (Experiment 2) and, in turn, this was more effective than water (Control). There was a high biofilm correlation (Pearson correlation) between both prostheses. Both products were well accepted by the patients, but the most favoured one was the paste. Conclusions: This was effective in controlling the biofilm and can be used preventatively in the maintenance of oral health by wearers of complete dentures. This is important where the lower prosthesis can harbour microorganisms which may act as a reservoir for other areas of the mouth and thus enhance the importance of proper hygiene.     

In situ evaluation of surface roughness and micromorphology of temporary soft denture liner materials at different time intervals

Objective

To perform an in situ evaluation of surface roughness and micromorphology of two soft liner materials for dentures at different time intervals.

Background

The surface roughness of materials may influence the adhesion of micro‐organisms and inflammation of the mucosal tissues. The in situ evaluation of surface roughness and the micromorphology of soft liner materials over the course of time may present results different from those of in vitro studies, considering the constant presence of saliva and food, the changes in temperature and the pH level in the oral cavity.

Materials and methods

Forty‐eight rectangular specimens of each of the two soft liner materials were fabricated: a silicone‐based material (Mucopren Soft) and an acrylic resin‐based material (Trusoft). The specimens were placed in the dentures of 12 participants (n = 12), and the materials were evaluated for surface roughness and micromorphology at different time intervals: 0, 7, 30 and 60 days. Roughness (Ra) was evaluated by means of a roughness tester. Surface micromorphology was evaluated by scanning electron microscopy.

Results

Analysis of variance for randomised block design and Tukey's test showed that surface roughness values were lower in the groups using the silicone‐based material at all the time intervals (P < .0001). The average surface roughness was higher at time interval 0 than at the other intervals, for both materials (P < .0001). The surface micromorphology showed that the silicone material presented a more regular and smoother surface than the acrylic resin‐based material.

Conclusion

The surface roughness of acrylic resin‐based and silicone‐based denture soft liner materials decreased after 7 days of evaluation, leading to a smoother surface over time. The silicone‐based material showed lower roughness values and a smoother surface than the acrylic resin‐based material, thereby making it preferred when selecting more appropriate material, due its tendency to promote less biofilm build‐up.     

Experiments on in vivo biofilm formation and in vitro adhesion of Candida species on polysiloxane liners

doi:10.1111/j.1741‐2358.2009.00329.x
Experiments on in vivo biofilm formation and in vitro adhesion of Candida species on polysiloxane liners Objectives: Microorganisms may colonise polysiloxane soft liners leading to bio‐deterioration. The aim of this study was to investigate in vitro adhesion and in vivo biofilm formation of Candida species on polysiloxane surfaces. Methods: The materials used in this study were Molloplast B, GC Reline soft, Mollosil Plus, Silagum Comfort and Palapress Vario. The in vitro retention of clinical isolates of Candida albicans to the relining and denture‐base materials by microscopic (scanning electron microscopy, SEM), conventional culturing methods and antimicrobial properties of these materials were studied. Candida found on materials and mucosa following long‐term use were identified and quantified, and biofilms covering the surfaces were investigated by SEM. Results: There was a significant decrease in the number of cells attached in vitro to saliva‐coated surfaces compared with non‐treated surfaces. An oral Candida carriage of 78% was found. Candida albicans, C. glabrata, C. intermedia and C. tropicalis were identified. In vivo biofilm formation on the liners appeared as massive colonisation by microorganisms. Conclusions: The results of the in vitro experiments suggest that salivary film influences early colonisation of different C. albicans strains. The film layer also minimises the differences among different strains. The Candida carriage of these patients was similar to denture‐wearing patients without soft liners.     

Effervescent tablets and ultrasonic devices against Candida and mutans streptococci in denture biofilm

doi: 10.1111/j.1741‐2358.2010.00378.x Effervescent tablets and ultrasonic devices against Candida and mutans streptococci in denture biofilm Objective: To evaluate the antimicrobial action of effervescent tablets and ultrasound on Candida spp. and mutans streptococci from denture biofilm. Background: It is not uncommon for edentulous patients to be elderly and find it difficult to brush their dentures. Hence, auxiliary methods are required for cleansing dentures as well as treating oral infections. Materials and methods: Seventy‐seven complete denture wearers were randomly assigned into four groups: (A) Brushing with water (control); (B) Effervescent tablets; (C) Ultrasonic device (Ultrasonic Cleaner, model 2840 D); (D) Effervescent tablets and ultrasonic device. All groups brushed their dentures with a specific brush and water, three times a day, before applying their treatments. Denture biofilm was collected at baseline and after 21 days. The samples were collected by brushing the dentures with saline and the detached microbial cells were quantified by plating. Counts [log (CFU+1) ml^?1] of total aerobes, Candida spp. and mutans streptococci were compared by one‐way anova or Kruskal–Wallis test (α = 0.05). Results: No significant difference was found among the methods from C. albicans (p = 0.76), C. tropicalis (p = 0.94) and C. glabrata (p = 0.80). Lower counts were found for methods B and D when compared with the other methods against mutans streptococci (p < 0.001). Method B showed lower total aerobic counts than A, whereas C and D showed intermediate results (p = 0.011). Conclusion: The effervescent tablets significantly reduced mutans streptococci and total aerobes from denture biofilm. However, they was not as effective against C. albicans. Ultrasonic cleansing presented a discrete antimicrobial effect and was less effective than the tablets for complete denture disinfection.     

Influence of varnish application on a tissue conditioner: analysis of biofilm adhesion

doi:10.1111/j.1741‐2358.2009.00290.x
Influence of varnish application on a tissue conditioner: analysis of biofilm adhesion Purpose: The aim of this study was to evaluate if the use of a varnish on a tissue conditioner would affect biofilm adhesion. Background: After the surgery has been performed, before the delivery of a complete denture, it is often necessary to use materials such as tissue conditioners on the surgical wound. However, these materials present deficient physico‐mechanical properties, which allow biofilm development. Methods: Forty elderly volunteers wearing complete maxillary dentures were selected. They were randomly allocated into two groups (n = 20), G1 and G2. In both groups, a silicone‐based tissue conditioner was placed in a recess created at the base of the denture, according to the manufacturer’s guidelines. In group G1, a varnish was applied to the tissue conditioner, while in group G2, no treatment was applied. All volunteers performed daily hygiene of the prosthesis with a soft toothbrush, and toothpaste. Quantification of the mean score values of biofilm formed at different time points (baseline, 1 week and 3 weeks) was statistically analysed using anova (α = 0.05). Results: Group G2 (1.6 ± 1.2) showed the lowest mean score values of biofilm formation and there was a statistical difference between the groups (p = 0.03). The highest mean score values were found after 3 weeks (2.7 ± 1.4) and were statistically different from the other time points studied (baseline and 1 week). Conclusion: The use of a varnish had a detrimental effect on the tissue conditioner studied, allowing higher biofilm formation.     

Complete denture hygiene and nocturnal wearing habits among patients attending the Prosthodontic Department in a Dental University in Brazil

Gerodontology 2010; doi: 10.1111/j.1741‐2358.2010.00369.x Complete denture hygiene and nocturnal wearing habits among patients attending the Prosthodontic Department in a Dental University in Brazil Objective: The aim of this study was to evaluate the overnight wearing and cleaning habits of complete denture wearers. Background: Successful complete denture treatment can be achieved when the patients are motivated and aware of appropriate denture wear and hygiene. Materials and methods: A sample of 224 complete denture wearers (162 women) aged 37–89 years was studied. Inclusion criteria comprised edentulous subjects who had received their new complete dentures between 2000 and 2005 in the Dental Clinic of the Araçatuba and Araraquara Dental School, São Paulo State University. Ethical approval was sought and granted. Subjects were interviewed using questions related to overnight denture wearing and denture cleaning habits. Possible statistical relationships among some of items were analysed by the chi‐square test at 5% significance level. Results: Of the patients, 55.8% removed their dentures during the overnight period and 88% did this every day. Among them, 66.4% removed both dentures. Most of the patients used brushing with toothpaste (105 patients – 46.87%) as a cleaning method. More than a half of the subjects (63.4%) showed biofilm and calculus on their dentures. Conclusion: The patients need instructions and motivation concerning denture hygienic and denture removal overnight.     

Evaluation of experimental cleanser solution of Ricinus communis: effect on soft denture liner properties

doi: 10.1111/j.1741‐2358.2010.00438.x Evaluation of experimental cleanser solution of Ricinus communis: effect on soft denture liner properties Objective: This study evaluated colour stability, hardness and roughness of soft denture liners after immersion in various cleansers. Materials and methods: Thirty specimens (14 mm × 4 mm) of Elite Soft Relining (ES) and Mucopren Soft (MS) were randomly immersed in distilled water at 37°C, sodium hypochlorite 1%, and an experimental Ricinus communis solution (RC) for 7, 15 and 183 continuous days. Results: anova (p < 0.05) and Tukey’s test indicated that after T7 (μ =8.79 ± 7.36); T15 (μ = 4.23 ± 2.62) and T183 (μ = 8.78 ± 3.16), MS presented a higher increase in hardness than ES. After T7, MS underwent an increase in roughness (μ = 0.09 ± 0.80); ES underwent a decrease (μ = ?0.08 ± 0.16). RC caused the smallest variation in roughness. After T15, both materials presented an increase in roughness. After T183, ES (μ = ?0.30 ± 0.48) presented a higher roughness variation than MS (μ = ?0.07 ± 0.32). Hypochlorite caused an increase in roughness (μ = 0.02 ± 0.19). Conclusion: After all periods ES presented higher colour alteration than MS; highest colour alteration was caused by hypochlorite. Both materials were more stable after immersion in RC.     

Distribution of biofilm on internal and external surfaces of upper complete dentures: the effect of hygiene instruction

Objectives: The purpose of this study was to evaluate biofilm distribution over internal and external surfaces of upper complete dentures. It was also aimed at assessing the effect of oral hygiene instructions before and after home use of a disclosing solution. Materials and methods: The sample consisted of 29 complete denture wearers whose upper prostheses were evaluated. Surface biofilm was disclosed by means of a 1% neutral red solution and quantified with regard to internal and external surfaces. Oral hygiene information was provided on an individual basis. During the second stage, an amount of disclosing solution was given for domestic use. Results: Internal and external surfaces presented a similar amount of biofilm, which was concentrated over the area of the posterior teeth area, palatal rugae and the internal vestibular incline of the distobuccal flange. This pattern was maintained during the study. However, overall amounts were reduced following denture hygiene information. The use of disclosing solution by subjects at home resulted in a further reduction. Conclusions: It was concluded that biofilm deposits were similar over assessed surfaces, regardless of the type of procedure but irregular areas presented greater amounts. Oral health instruction was effective in reducing the biofilm, in particular when associated with home use of a disclosing agent.     

Complete denture wearing and fractures among edentulous patients treated in university clinics

doi: 10.1111/j.1741‐2358.2011.00551.x
Complete denture wearing and fractures among edentulous patients treated in university clinics Objective: The prevalence of wearing and fracture of complete dentures was evaluated among edentulous patients treated in two dental schools in Brazil. Background: Acceptance and wearing of complete dentures are related to adaptive behaviour of edentulous patients. However, one reason that could interfere with the wearing dentures is their potential to fracture, which is still a common complication in denture rehabilitation practice. Material and methods: Two hundred and twenty‐four edentulous patients rehabilitated with complete dentures from 2000 to 2005 in Araçatuba and Araraquara Dental School, University of State of São Paulo, were assessed in 2006 and 2007 to answer a questionnaire about wearing and fracture of their dentures. Statistical analysis were performed using Epi Info software and chi‐squared test to compare maxillary and mandibular data (α = 0.05). Results: Almost 26% of the patients did not wear their dentures, and among the remainder, the majority wore the maxillary denture. About 30% of the dentures were fractured, with higher prevalence in the maxillary arch (p = 0.003). Conclusions: Discontinuation of wearing dentures was quite high, especially considering the treatment which was carried out in university clinics. Prevalence of fractures was also high, greater for the maxillary denture, and was one of the main reasons for non‐wearing of complete dentures.     

Evaluation of three indices for biofilm accumulation on complete dentures

doi:10.1111/j.1741‐2358.2009.00285.x
Evaluation of three indices for biofilm accumulation on complete dentures Objectives: The objective of this study was to evaluate the accuracy and reproducibility of three complete denture biofilm indices (Prosthesis Hygiene Index; Jeganathan et al. Index; Budtz‐Jørgensen Index) by means of a computerised comparison method. Background: Clinical studies into denture hygiene have employed a large number of biofilm indices among their outcome variables. However, the knowledge about the validity of these indices is still scarce. Materials and methods: Sixty‐two complete denture wearers were selected. The internal surfaces of the upper complete dentures were stained (5% erythrosine) and photographed. The slides were projected on paper, and the biofilm indices were applied over the photos by means of a scoring method. For the computerised method, the areas (total and biofilm‐covered) were measured by dedicated software (Image Tool). In addition, to compare the results of the computerised method and Prosthetic Hygiene Index, a new scoring scale (including four and five graded) was introduced. For the Jeganathan et al. and Budtz‐Jørgensen indices, the original scales were used. Values for each index were compared with the computerised method by the Friedman test. Their reproducibility was measured by means of weighed κ. Significance for both tests was set at 0.05. Results: The indices tested provided similar mean measures but they tended to overestimate biofilm coverage when compared with the computerised method (p < 0.001). Agreement between the Prosthesis Hygiene Index and the computerised method was not significant, regardless of the scale used. Jeghanathan et al. Index showed weak agreement, and consistent results were found for Budtz‐Jorgensen Index (κ = 0.19 and 0.39 respectively). Conclusion: Assessment of accuracy for the biofilm indices showed instrument bias that was similar among the tested methods. Weak inter‐instrument reproducibility was found for the indices, except for the Budtz‐Jørgensen Index. This should be the method of choice for clinical studies when more sophisticated approaches are not possible.     

In vivo assessment of the effect of an adhesive for complete dentures on colonisation of Candida species

doi: 10.1111/j.1741‐2358.2009.00345.x
In vivo assessment of the effect of an adhesive for complete dentures on colonisation of Candida species Objective: Denture adhesives have long been recognised by denture wearers as a useful adjunct to denture retention and stability. The objective of the present study was to evaluate, in vivo, the effect of a denture adhesive on oral quantities of Candida species by determination of absolute counts of colony‐forming units (CFU) per ml of saliva of individuals who use this denture adhesive for a period of 14 days. Materials and methods: Twenty‐four individuals were randomised in two equal groups of 12 (test and control), with the individuals of the test group using the adhesive for 14 days. Samples of saliva were collected from all individuals on days 0 (initial), 7 and 14. Aliquots of saliva were diluted and plated in duplicate on Sabouraud dextrose agar with chloramphenicol and incubated for 37°C for 48 h, the CFU/ml were counted in the individuals of each group and the data of each group were compared at the different time periods and analysed statistically by the non‐parametric Mann‐Whitney U‐test (α ≤ 5%). Results: There were no statistically significant differences between the test and control groups during the test periods. Conclusion: Within of the limitations of this study, the data suggested that the denture adhesive tested did not significantly alter the oral microbiota during the 14‐day trial period.     

Relining of prosthesis with auto‐polymerizing hard denture reline resins: effect of post‐polymerization treatment on flexural strength

Background: It has been suggested that microwave irradiation and prosthesis immersion in hot water after its polymerization may improve mechanical and viscoelastic properties of acrylic resins. Purpose: This study was proposed to verify the influence of microwave post‐polymerization (PP) treatment over the flexural strength of thermo‐polymerizing acrylic resin specimens (QC‐20) relined or not with two different composition hard chairside auto‐polymerizing reliners [Kooliner (K) and New Truliner (NT)]. Materials and Methods: For this study, 50 specimens of 64 × 10 × 3.3 mm were polymerized and distributed into five groups. G1 (control) specimens without relining and PP; G2 specimens relined with K, without PP; G3 specimens relined with NT, without PP; G4 specimens relined with K, with PP (microwave irradiation with 650 W for 5 min); G5 specimens relined with NT, with PP. Tests were performed on a universal testing machine Instron 4411 with compression speed of 5 mm/min. Results: Specimens of K without PP did not show statistically different results (p < 0.05) when compared with control. However, when submitted to PP these specimens showed a significant increase in flexural strength. Specimens of NT showed the lowest flexural strength of all groups, with or without PP when compared with control and K groups. Conclusion: Microwave PP (650 W for 5 min) proved to be an effective method of improving the flexural strength of K relined prosthesis. However, it did not seem to affect NT specimens.     

Effect of denture adhesive on the micro-organisms in vivo

doi: 10.1111/j.1741‐2358.2010.00381.x Effect of denture adhesive on the micro‐organisms in vivo Background: Denture adhesives increase the retention and stability of dentures in edentulous patients, especially in cases where salivary flow is impaired or in the management of traumatised oral mucosa. Objectives: The effect of a denture adhesive on the oral flora at different time intervals. Method: Thirty denture‐wearing patients were involved in this study. While half of the group received a denture adhesive, the other half did not. At baseline, 1 and 2 months after delivering the dentures, smear samples were obtained from the saliva, palate and the dentures. Candida albicans, Candida krusei, Candida glabrata, Candida spp., Staphylococcus aureus, Moraxella catarrhalis, α‐haemolytic streptococci, β‐haemolytic streptococci, Pneumococcus aureus, S. anginosus, S. intermedius, S. constellatus, S. sanguis, S. gordonii, S. mitis, S. mutans, S. salivarius, and yeasts were investigated. The data were statistically analysed using anova and repeated measures. Results: Most types of the micro‐organisms were not seen and could not be analysed statistically except α‐haemolytic streptococci and C. albicans. No statistically significant difference was found for α‐haemolytic streptococci and C. albicans in saliva, palate and the denture at all time intervals. Conclusions: Prolonged use of the denture adhesive tested up to 2 months did not yield to increase in micro‐organisms of the oral flora.