Impact of Denture Cleaning Method and Overnight Storage Condition on Denture Biofilm Mass and Composition: A Cross-Over Randomized Clinical Trial

Background

Appropriate oral hygiene is required to maintain oral health in denture wearers. This study aims to compare the role of denture cleaning methods in combination with overnight storage conditions on biofilm mass and composition on acrylic removable dentures.

Methods

In a cross-over randomized controlled trial in 13 older people, 4 conditions with 2 different mechanical cleaning methods and 2 overnight storage conditions were considered: (i) brushing and immersion in water without a cleansing tablet, (ii) brushing and immersion in water with a cleansing tablet, (iii) ultrasonic cleaning and immersion in water without a cleansing tablet, and (iv) ultrasonic cleaning and immersion in water with a cleansing tablet. Each test condition was performed for 5 consecutive days, preceded by a 2-days wash-out period. Biofilm samples were taken at baseline (control) and at the end of each test period from a standardized region. Total and individual levels of selected oral bacteria (n = 20), and of Candida albicans were identified using the Polymerase Chain Reaction (PCR) technique. Denture biofilm coverage was scored using an analogue denture plaque score. Paired t-tests and Wilcoxon-signed rank tests were used to compare the test conditions. The level of significance was set at α< 5%.

Results

Overnight denture storage in water with a cleansing tablet significantly reduced the total bacterial count (p<0.01). The difference in total bacterial level between the two mechanical cleaning methods was not statistically significant. No significant effect was observed on the amount of Candida albicans nor on the analogue plaque scores.

Conclusions

The use of cleansing tablets during overnight denture storage in addition to mechanical denture cleaning did not affect Candida albicans count, but reduced the total bacterial count on acrylic removable dentures compared to overnight storage in water. This effect was more pronounced when combined with ultrasonic cleaning compared to brushing.

Trial Registration

ClinicalTrials.gov {"type":"clinical-trial","attrs":{"text":"NCT02454413","term_id":"NCT02454413"}}NCT02454413     

Effectiveness of different cleaning agents on the adherence of Candida albicans to acrylic denture base resin

doi: 10.1111/j.1741‐2358.2010.00379.x
Effectiveness of different cleaning agents on the adherence of Candida albicans to acrylic denture base resin Objective:  To evaluate the ability of three alkaline peroxide‐type (Polident, Efferdent, Fittydent) and two mouth rinse cleaning agents (CloSYSII and Corsodyl) to inhibit Candida albicans on acrylic denture base resin. Background:  Appropriate routine cleaning of dentures is necessary to prevent denture stomatitis and maintenance of healthy supporting tissues. Materials and methods:  A total of 180 acrylic resin specimens (10 × 10 × 2 mm) were prepared and divided into six groups. Candida albicans was incubated on Sabouraud dextrose agar (SDA) at 37°C for 48 h. After dilution, a final yeast suspension of approximately 10 6 C. albicans per millimetre was prepared. Ten acrylic resin specimens for each group were placed in a sterile Petri dish covered with 20 ml of fungal suspension and incubated at 37°C for 90 min. Then, the specimens were immersed in 40 ml of the test solution at 37°C for 15, 30 and 60 min. Fungal cells adhering to acrylic resin surfaces were fixed in formaldehyde and counted microscopically. Results:  Mouth rinses showed the highest removal activity for all the treatment times and completely eliminated the adherence of C. albicans. Conclusions:  The use of mouth rinse may be a suitable method for cleaning dentures.     

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.     

Factors affecting the attachment of micro-organisms isolated from ultrafiltration and reverse osmosis membranes in dairy processing plants

Aims:  To identify the types of micro-organisms involved in the formation of biofilms on dairy ultrafiltration and reverse osmosis membranes and investigate factors affecting the attachment of those isolates.
Methods and Results:  Micro-organisms isolated from industrial membranes following standard cleaning were identified using the API culture identification system. Thirteen different isolates representing eight genera were isolated and their ability to attach to surfaces was compared using a microtitre plate assay. Three Klebsiella strains attached best, while mixed strains of Pseudomonas and Klebsiella attached better than individual strains. Whey enhanced the attachment of the isolates. The micro-organisms were characterized according to cell surface hydrophobicity using the microbial adhesion to hydrocarbon (MATH) test, and cell surface charge by measuring the zeta potential. These cell surface characteristics did not show a clear relationship with the attachment of our strains.
Conclusions:  A variety of different micro-organisms is associated with dairy ultrafiltration and reverse osmosis membranes after cleaning, suggesting several possible sources of contamination. The cleaning of these membranes may be inadequate. The attachment of the different isolates is highly variable and enhanced in the presence of whey.
Significance and Impact of the Study:  Knowledge of persistent microflora colonizing dairy membrane systems will help develop strategies to mitigate biofilm development in this environment, improving hygiene in membrane processing plants.     

Anti-adhesive and antibacterial properties of a proprietary denture cleanser

The antimicrobial efficacy and anti-adhesive attributes of a proprietary denture cleanser were evaluated. To determine cleanser antimicrobial efficacy, Streptococcus mutans was grown on denture acrylic strips which were then exposed to the cleanser. To evaluate anti-adhesive efficacy, the strips were treated with the cleanser and then placed in the Strep. mutans suspension. Following incubation, adhered bacteria were removed and enumerated by viable counting. Treated denture acrylic plates were also placed in a parallel-plate flow chamber and then exposed to Strep. oralis. Images of adhered bacteria were analysed to determine biofilm coverage. Biofilm removal force was quantified by increasing the flow rate between the acrylic plates. The cleanser exhibited a 100% kill against Strep. mutans adhered to the acrylic surface, and inhibited attachment of cells by 66%. The flow chamber study found that cleanser-treated denture acrylic allowed the formation of a multilayer biofilm which was easily removed by a slight increase in flow rate.     

Survival analysis of mandibular complete dentures with acrylic‐based resilient liners

doi: 10.1111/j.1741‐2358.2012.00658.x Survival analysis of mandibular complete dentures with acrylic‐based resilient liners Objective: The purpose of this long‐term randomised controlled trial was to compare the longevity of dentures constructed using a conventional acrylic resin (CAR) to that of dentures constructed using an acrylic‐based resilient liner (ARL). Materials and methods: The follow‐up study was essentially carried out by annual telephone calls to each of the 67 participants. The Kaplan–Meier method and life‐table analysis were used for univariate analyses. The Cox proportional‐hazards test was used as a final model for statistically adjusting predictor variables such as sex, clinician type, mandibular denture type and age at denture delivery. Results: The denture type was likely to affect the survival time of the dentures, while the sex and clinician type were not. The group using acrylic‐based resilient denture liners had twice the risk of having shorter denture‐survival times than those using conventional acrylic resin dentures. Younger participants were likely to have a reduced risk of having shorter denture‐survival times than older participants. Conclusion: We conclude that mandibular complete dentures constructed using ARL are twice as likely as dentures constructed using CAR to have shorter denture survival times, mainly because of material deterioration.     

Examination of denture-cleaning methods based on the quantity of microorganisms adhering to a denture

doi: 10.1111/j.1741‐2358.2011.00461.x Examination of denture‐cleaning methods based on the quantity of microorganisms adhering to a denture Objectives: To investigate effective denture‐cleaning methods, we examined the relationships between the quantity of microorganisms adhering to dentures and the use of a denture brush and the frequency of use of a denture cleanser. Subjects and Methods: Denture plaque was collected from the mucosal surface of the examined dentures, which were 142 and 80 upper and lower complete dentures, respectively, worn by 96 outpatients (mean age: 71.9 years) of a university hospital and 41 nursing home residents (mean age: 84.8 years). The collected microorganisms were counted in terms of isolated representative colonies that were cultured and identified using standard methods. The use of a denture brush, the frequency of use, and the type and soaking time of denture cleansers as denture‐cleaning methods were surveyed. Results: The quantity of microorganisms was significantly lower in dentures of denture brush users than in those of non‐users in the outpatients (p < 0.01, Mann–Whitney U test). The quantity of microorganisms was significantly lower in the dentures of outpatients who used a denture cleanser daily or 3–4 times a week than in those who used one once or less per month and in the dentures of nursing home residents who used one daily than in those who used one at other frequencies (p < 0.05, Kruskal–Wallis test, followed by Dunn’s Multiple Comparison test). Conclusion: Within the limitations of this study, it was concluded that the use of a denture brush and daily use of denture cleanser should be recommended to complete dentures wearers as denture‐cleaning methods that effectively reduce the quantity of microorganisms adhering to dentures.     

High correlation between the bacterial species in denture plaque and pharyngeal microflora

Objectives : The morbidity and mortality of the dependent elderly that result from aspiration pneumonia are recognized as a major geriatric health problem. Most cases of bacterial pneumonia are initiated following colonization or superinfection of the pharynx by pathogenic bacteria, followed by aspiration of pharyngeal contents. A recent study revealed that bacteria, that commonly cause respiratory infection, colonized the dentures of dependent elderly. This suggests that denture plaque may function as a reservoir of potential respiratory pathogens to facilitate colonization on the pharynx. The purpose of this study was to determine the possible correlation between denture and pharyngeal microflora. Study Design : The denture and pharyngeal bacterial flora of 50 dependent elderly were examined, and the microorganisms identified by culturing. The agreement between the bacterial species in denture plaque and pharyngeal microflora was investigated using the Kappa method. Results : The microorganism species on the dentures and pharyngeal mucosa of the subjects had an agreement rate of 68.5%. The agreement rate for each of the bacterial species of the dentures and pharynx was also demonstrated to be high. Conclusions : Dentures should be considered an important reservoir of organisations which could colonise the pharynx, and the importance of controlling denture plaque for the prevention of aspiration pneumonia cannot be overemphasized.     

The effects of surface roughness and type of denture acrylic on biofilm formation by Streptococcus oralis in a constant depth film fermentor

AIMS: To investigate the effects of surface roughness and type of denture acrylic on the early development of a Streptococcus oralis biofilm in a constant depth film fermentor (CDFF). METHODS AND RESULTS: Streptococcus oralis was incubated with acrylic of known surface roughness in the CDFF. Adherent Strep. oralis were enumerated by viable counting. Cold-cure acrylic was rougher (P < 0.01) than heat-cure acrylic after polishing with abrasive paper of any given grit-grade. Heat-cure acrylic was colonized by fewer (P < 0.001) bacteria than cold-cure acrylic at any given surface roughness. The number of bacteria adhering to heat-cure and cold-cure acrylic increased linearly with mean surface roughness after 2 h incubation, the increase being greater (P < 0.001) for the cold-cure compared with the heat-cure acrylic. However, after 4 h incubation, surface roughness appeared to have no effect on the number of adherent bacteria. CONCLUSION: The type of acrylic used, and its roughness, affect the early stages of biofilm formation by Strep. oralis. SIGNIFICANCE AND IMPACT OF THE STUDY: Choosing an appropriate type of smooth acrylic could lead to reduced biofilm formation in vivo.     

The needs of denture-brushing in geriatrics: clinical aspects and perspectives

doi: 10.1111/j.1741‐2358.2011.00557.x
The needs of denture‐brushing in geriatrics: clinical aspects and perspectives Introduction: Oral and denture hygiene are often defective in particular with dependent persons such as geriatric subjects. The reasons are the lack of hygiene education of the subjects or those caring for them. Consequently, oral hygiene is often neglected, resulting in poor oral health and an increase in the presence of local or general infections. Objective: This paper is a report of brushing effectiveness on microbial biofilm deposits on dentures of subjects participating in a specific oral hygiene programme. Materials and methods: Thirty‐nine dentures of 30 subjects were assessed for 2 weeks following an educational brushing programme. Microbial biofilm was recovered using fluoresceine and then scanned and quantified by ‘Mesurim’ software three times: before study, after 1 and 2 weeks. Results: The repeated measurement procedures showed a decrease in the percentage of biofilm present (F = 15, p < 0.001) whatever the type of denture (partial or complete) and for all biomaterials. Conclusion: Regular denture‐brushing can improve local hygiene. Consequently, decreasing the biofilm surface can reduce the prevalence of oral pathogens, thereby contributing to the general prevention of the risks of infections such as pneumotisis.     

Effect of sealer coating on mechanical and physical properties of permanent soft lining materials

doi: 10.1111/j.1741‐2358.2011.00487.x
Effect of sealer coating on mechanical and physical properties of permanent soft lining materials Objective: To evaluate the long‐term effects of sealer coating on tensile bond strength and surface roughness of soft liners. Background: Failure of the bond between resilient liners and denture base significantly compromise the dentures’ longevity. In addition, surface roughness contributes to bacterial adherence on prosthetic materials, increasing the risk of oral infections. Methods: Specimens were manufactured from four reliners [Mucopren Soft (MS), Dentuflex (DF); Soft Comfort Denso (SC) and Ufi Gel SC (USC)], distributed into 10 groups (n = 10), according to material and coating treatment. Tensile bond strength was performed after one year of ageing, while surface roughness was evaluated at 0, 1, 3, 6 and 12 ageing months. Bond strength data were submitted to two‐way anova and Tukey‐HSD tests, while roughness data were submitted to mixed model analysis for repeated measurements (p ≤ 0.05). Results: MS and DF without sealer coating presented the highest tensile bond strength. After coating, DF and SC presented increased tensile bond strength. No surface roughness difference was observed in the non‐coated groups over time, acrylic‐based reliners presented higher roughness. Among coated groups, only SC presented increased surface roughness. Acrylic‐based materials presented reduced roughness at three months. Conclusion: Surface coating was effective for acrylic reliners in maintaining their initial properties. However, sealer coating should be re‐applied every 3 months.     

Fracture resistance of maxillary complete dentures subjected to long-term water immersion

doi: 10.1111/j.1741‐2358.2012.00616.x Fracture resistance of maxillary complete dentures subjected to long‐term water immersion Objective: This study investigated the fracture resistance of maxillary acrylic resin complete dentures subjected to long‐term water immersion. Materials and Methods: Maxillary acrylic resin complete dentures were fabricated from five denture base resins. Half of the dentures were stored in water for 50 h, and the other half were kept in water for 180 days before testing. Ten specimens were fabricated per group. The flexural load at the proportional limit (FL‐PL) of the dentures was tested. Results: A two‐way anova revealed a significant difference in FL‐PL because of the denture base material variable. There were no significant differences in FL‐PL because of the effect of water immersion and the interaction between the effect of water immersion and the denture base material. The FL‐PLs of the dentures fabricated with the two conventional heat‐processed resins, the pour‐type autopolymerizing resin and the microwave energy‐processed resin were not significantly different from each other; they were significantly higher than the light‐activated resin in regard to their FL‐PL. Conclusion: The FL‐PLs of the maxillary acrylic resin complete dentures did not change after long‐tern water immersion, and the FL‐PL of the denture fabricated from the light‐activated resin was lower than those of the other materials.     

Weight loss and changes in surface roughness of denture base and reline materials after simulated toothbrushing in vitro

doi: 10.1111/j.1741‐2358.2010.00422.x
Weight loss and changes in surface roughness of denture base and reline materials after simulated toothbrushing in vitro Objective: To evaluate the weight loss and the surface roughness of acrylic resins after simulated brushing tests. Material and methods: Ten specimens of each material (Tokuyama Rebase II‐TR, New Truliner‐NT, Ufi Gel Hard‐UH and Lucitone 550‐L) were made. The wear loss (mg) by weight and the surface roughness (Ra μm) of each specimen was determined before and after brushing. The specimens were placed on the brushing machine and a total of 20 000 brushing cycles was performed. The results of weight loss and roughness values were submitted to the anova followed by the Tukey’s test (p = 0.05). Results: The mean weight loss of material L was statistically higher (p < 0.001) than that of the relines TR, UH and NT. No significant differences were found among the roughness values of resins TR, UH and L (p > 0.05). Only for L, toothbrushing increased the surface roughness. After toothbrushing, there was no significant difference between the roughness values of materials L and NT. The highest mean weight loss during the simulated toothbrushing tests was observed for L. Before the toothbrushing tests, the NT exhibited the highest mean roughness. Conclusion: Brushing resulted in increase in roughness only for resin L.     

Correlation between factors associated with the removable partial dentures use and Candida spp. in saliva

doi: 10.1111/j.1741‐2358.2010.00390.x
Correlation between factors associated with the removable partial dentures use and Candida spp. in saliva Objectives: To correlate the presence and number of Candida spp. in the saliva of wearers of removable partial dentures retained with precision attachments with the proportion of metal/acrylic resin present in the dentures. Methods: Saliva samples from 40 removable partial denture wearers (test) and one paired sample of individuals, non‐wearers of any type of removable denture (control) were collected, seeded, and the colony forming units of Candida counted and identified. The metal/acrylic resin proportion of each denture was quantified, using silicone plates pressed over each denture. Results: Candida spp. was found in the saliva of 80% of the individuals in the test group and 65% of the control, with C. albicans being the most prevalent species. The test group presented with the highest number of colony forming units of Candida per ml of saliva, and there was weak correlation between this number and the metal and resin area of the denture (Pearson’s coefficient of correlation). Greater prevalence and a higher number of colony forming units of Candida per ml of saliva occurred in removable partial denture wearers (p = 0.04) with a weak positive correlation between the metal and resin area and the number of colony forming units of Candida per ml of saliva. However, this correlation was more significant for the area of resin.     

Susceptibility of MRSA biofilms to denture-cleansing agents

Methicillin-resistant Staphylococcus aureus (MRSA) is an important nosocomial pathogen, which is responsible for considerable morbidity and mortality in the United Kingdom. The major reservoir of this organism is thought to be the anterior nares, but there is increasing evidence that this pathogen is present in the oral cavity, particularly in denture wearers. The purpose of this study was to determine whether MRSA, grown as biofilms on denture acrylic resin, could be eradicated using commercially available agents. EMRSA-15 or EMRSA-16 was grown in a model system on the surface of denture acrylic resin for 4, 24 or 120 h before the samples were exposed to a range of disinfectants for time intervals of 1, 5 and 10 min. All of the agents reduced the number of cultivable MRSA bacteria present on the acrylic resin surface at 4 h, with 2% sodium hypochlorite (NaOCl) eliminating MRSA below the level of detection after an exposure of 1 min. However, the established MRSA biofilms (24 and 120 h) were more resistant to killing by the agents, although 2% NaOCl was still able to eradicate all ages of MRSA biofilms within 1 min of exposure.     

Model systems allowing quantification of sensitivity to disinfectants and comparison of disinfectant susceptibility of persistent and presumed nonpersistent Listeria monocytogenes

Aims:  To determine if Listeria monocytogenes persistent strains differ from presumed nonpersistent strains in disinfection susceptibility and to examine the influence of attachment and NaCl on susceptibility.
Methods and Results:  Two model-systems that allowed quantitative inter-strain comparison of disinfectant sensitivity were developed. Persistent L. monocytogenes were not more tolerant to the disinfectants Incimaxx DES and Triquart SUPER than presumed nonpersistent isolates. When calibrating the systems with respect to presence of biological material and cell density, attached bacteria were as sensitive to disinfectants as were planktonic bacteria. Growth with 5% NaCl increased the tolerance of planktonic cells to Incimaxx DES. All strains of spot inoculated L. monocytogenes survived well 20 h of drying when protected by growth media and 5% NaCl, but were not protected by NaCl against disinfection.
Conclusions:  Persistent strains of L. monocytogenes are as susceptible to disinfectants as are presumed nonpersistent strains and attachment does not render the strains more tolerant to disinfectants. Growth with NaCl affected the susceptibility of the planktonic L. monocytogenes to Incimaxx DES and protected spot inoculated cells during drying.
Significance and Impact of the Study:  Attachment to surfaces does not per se offer protection to L. monocytogenes against disinfectants and disinfection tolerances do not appear to influence the ability of a strain to persist.     

Bacteria biofilm encourages algal immigration onto substrata in lotic systems

I tested the effect of the density of attached bacteria on the amount of algal immigration in the early development of a periphyton community in an artificial stream by manipulating the density of the attached bacteria. Three densities were prepared by regulation of the incubation time. A suspension of algae was added to the stream, and the degree of algal attachment to substrata was compared among the treatments. Algal immigration was proportional to the density of attached bacteria on all substrata (glass, PVC, and slate), although density differed among substrata. Analysis of covariance (dependent variable, amount of attached algae; covariate, bacterial density) showed significant relationship between amounts of attached algae and bacterial densities, but did not show significant differences in the slopes and adjusted means among substrata. When acrylic beads were added with the suspension of attached algae, significant linear correlation was obtained between the amount of attached algae and the amount of acrylic beads on the substrata. Algal immigration was due to non-selective adsorption by attached bacterial biofilms on substrata, although the extent of bacterial colonization and biofilm formation may be affected by the substrata and other environmental factors (e.g., current conditions and water temperature).     

Enhanced biofilm formation and 3-chlorobenzoate degrading activity by the bacterial consortium of Burkholderia sp. NK8 and Pseudomonas aeruginosa PAO1

Aims:  To characterize biofilm formation of a chlorobenzoates (CBs) degrading bacterium, Burkholderia sp. NK8, with another bacterial species, and the biodegradation activity against CBs in the mixed-species biofilm.
Methods and Results:  Burkholderia sp. NK8 was solely or co-cultured with each of five other representative bacteria in microtitre dishes. Biofilm formation involving the strain NK8 was synergistically promoted by co-culturing with only Pseudomonas aeruginosa PAO1. Epifluorescent microscopy revealed that cells of the bacterial strain NK8 were viable and distributed randomly in the mixed-species biofilms. Enumeration of the attached cells on the surface of wells revealed that cells of the strain NK8 increased approx. 10-fold by the co-culture with the strain PAO1 compared to those by monoculture of the strain NK8, and the degradation activity of 3-chlorobenzoate by the dual-species biofilms was more promoted than that by the strain NK8-monocultured biofilms.
Conclusions:  Enhanced biofilm formation of Burkholderia sp. NK8 by the bacterial consortium occurred, but is determined by the partner bacterial species. The mixed-species biofilms have the advantage to degrade CBs on a solid surface.
Significance and Impact of the Study:  This study provides a significance of bacterial consortia on the biofilm formation and the degradation activity of Burkholderia sp. NK8, which contribute for complete degradation of chlorinated aromatics.