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.     

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.     

Virulence and pathogenicity of Candida albicans is enhanced in biofilms containing oral bacteria

This study examined the influence of bacteria on the virulence and pathogenicity of candidal biofilms. Mature biofilms (Candida albicans-only, bacteria-only, C. albicans with bacteria) were generated on acrylic and either analysed directly, or used to infect a reconstituted human oral epithelium (RHOE). Analyses included Candida hyphae enumeration and assessment of Candida virulence gene expression. Lactate dehydrogenase (LDH) activity and Candida tissue invasion following biofilm infection of the RHOE were also measured. Candida hyphae were more prevalent (p < 0.05) in acrylic biofilms also containing bacteria, with genes encoding secreted aspartyl-proteinases (SAP4/SAP6) and hyphal-wall protein (HWP1) up-regulated (p < 0.05). Candida adhesin genes (ALS3/EPA1), SAP6 and HWP1 were up-regulated in mixed-species biofilm infections of RHOE. Multi-species infections exhibited higher hyphal proportions (p < 0.05), up-regulation of IL-18, higher LDH activity and tissue invasion. As the presence of bacteria in acrylic biofilms promoted Candida virulence, consideration should be given to the bacterial component when managing denture biofilm associated candidoses.     

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.     

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.     

Genetic diversity of gamma-hexachlorocyclohexane-degrading sphingomonads isolated from a single experimental field

Aim:  To determine the effect of the surface roughness of denture acrylic on the attachment of Streptococcus oralis .
Methods and Results:  Roughened denture acrylic samples were assessed for bacterial attachment, over time, using microscopy. The area of the image covered by bacteria was calculated and converted into a percentage of the total area sampled. The results showed an increasing bacterial coverage with time of incubation and increasing roughness. Differences were seen between heat cured acrylic and cold cured acrylic.
Conclusion:  This study successfully demonstrated a system for the assessment of the amount of attached bacteria on denture acrylic varying roughness. The system was able to discern the difference in surface area coverage by attached bacteria over a roughness range relevant to brushing dentures with dentifrices.
Significance and Impact of Study:  This study provides strong support for the scratches caused by brushing dentures with dentifrice encouraging bacterial attachment. This is likely to have a significant effect on efficacy of denture cleaning, general hygiene and biofilm re-formation between cleaning regimens and may indicate that alternative low abrasive cleaners, such as antimicrobial denture-cleaning tablets, offer a more appropriate regimen.     

Evaluation of Candida albicans adhesion and biofilm formation on a denture base acrylic resin containing silver nanoparticles

Aim: This study firstly evaluated the activity of a silver nanoparticle (AgNPs) solution against Candida albicans and then the effect of incorporation of AgNPs into a denture base acrylic resin on the material’s hydrophobicity, C. albicans adhesion and biofilm formation. Methods and Results: The AgNPs solution was synthesized by chemical reduction and characterized. Minimum inhibitory (MIC) and minimum fungicidal (MFC) concentrations for planktonic cells and sessile cells (MFCs) of the AgNPs solution against C. albicans were determined. Specimens (n = 360) of silver‐incorporated acrylic resin at concentrations of 1000, 750, 500, 250 and 30 ppm were also prepared and stored in PBS for 0, 7, 90 and 180 days. Control was acrylic resin without AgNPs (0 ppm). After the storage periods, contact angles were measured and the specimens were used for C. albicans adherence (37°C; 90 min; n = 9) and biofilm formation (37°C; 48 h; n = 9) by XTT reduction assay. MIC, MFC and MFCs values were 3·98, 15·63 and 1000 ppm, respectively. Incorporation of AgNPs reduced the hydrophobicity of the resin. No effect on adherence and biofilm formation was observed. At 90 and 180 days of storage, there was significant increase in adherence and biofilm formation. Conclusions: Although the AgNPs solution had antifungal activity, no effect on C. albicans adherence and biofilm formation was observed after its incorporation into a denture base resin. Significance and Impact of the Study: The synthesized AgNPs solution is a promising antifungal agent, warranting investigations of more efficient methods of incorporation into denture base resins.     

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.     

In vivo biofilm formation on a soft denture liner in elderly patients with controlled diabetes

doi: 10.1111/j.1741‐2358.2010.00428.x In vivo biofilm formation on a soft denture liner in elderly patients with controlled diabetes Objectives: This in vivo study evaluated the influence of controlled diabetes on biofilm formation on a soft denture liner in elderly patients. Background: Soft denture lining materials are more susceptible to microbial colonisation than denture base acrylic resins. Especially in the elderly, several predisposing factors may accumulate leading to an increased probability of biofilm development that may result in candidiasis, a significant clinical oral disease. Materials and methods: Volunteers wearing complete dentures were divided into two groups (n = 20): diabetic patients with controlled glycaemia, and healthy patients. In both groups, a silicone‐based soft liner was placed in a recess created at the base of the maxillary dentures. Subjects cleaned the prosthesis three times a day. Biofilm formed on the liner was quantified at various time points (baseline, two, four and six weeks). Data were analysed by two‐way repeated measures anova and Tukey’s test (α = 0.05). Results: There was no statistical difference in biofilm formation for any of the time points between controlled diabetes patients and healthy patients. Conclusion: The results suggest that the control of diabetes in elderly patients provides the same levels of biofilm formation when compared to healthy individuals.     

Inhibition of Streptococcus pyogenes Biofilm Formation by Coral-Associated Actinomycetes

Streptococcus pyogenes biofilms tend to exhibit significant tolerance to antimicrobials during infections. We screened coral-associated actinomycetes (CAA) for antibiofilm activity against different biofilm forming M serotype of Streptococcus pyogenes. Actinomycetes isolated from the mucus of the coral Acropora digitifera were screened for antibiofilm activity against S. pyogenes biofilms wherein several isolates clearly demonstrated antibiofilm activity. The biofilm inhibitory concentrations (BICs) and the sub-BICs (1/2 and 1/4 BIC) of the extracts significantly prevented biofilm formation up to 60–80%. The extract of Streptomyces akiyoshinensis (A3) displayed efficient antibiofilm activity against all the biofilm forming M serotypes. All the five extracts efficiently reduced the cell surface hydrophobicity (a crucial factor for biofilm formation in S. pyogenes) of three M types and thus may inhibit biofilm formation. CAA represent an interesting source of marine invertebrates-derived antibiofilm agents in the development of new strategies to combat Streptococcal biofilms.     

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.     

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.     

Effect of pre-processing methods on bond strength between acrylic resin teeth and acrylic denture base resin

doi: 10.1111/j.1741‐2358.2011.00480.x
Effect of pre‐processing methods on bond strength between acrylic resin teeth and acrylic denture base resin Objectives: This study evaluated the effects of various pre‐processing methods on the bond strength between resin and denture teeth. Backgrounds: Debonding of acrylic resin teeth from denture base material is a problem for patients wearing complete dentures. Materials and Methods: Four experimental groups (n = 30) were investigated by subjecting tooth–resin bonding to tensile loading. Specimens were prepared and tested according to the methods of the International Standards Organization (ISO 22112:2005) using a special assembly. Four pre‐processing surface treatments of teeth were applied: (i) ST₁, no treatment applied (control); (ii) ST₂, wax solvent (Dewaks, Faber Kimya & Ilaç, Turkey); (iii) ST₃, boiling water followed by conditioning with methyl methacrylate (MMA) monomer (Meliodent, Bayer Dental, Germany); (iv) ST₄, boiling water followed by wax solvent agent and finally MMA monomer application. Bond strength test was performed using a universal testing machine. Results: All the strength values of the test groups were within clinically acceptable limits. The lowest values were from the ST₁ group and the highest values were in the ST₄ group. Conclusions: Wax elimination methods affected bonding strength. Application of wax solvent and MMA monomer to the ridge lap surfaces of the teeth gave the best results. In clinical practice, this application procedure may decrease the bonding failure of denture teeth.     

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.     

Bond strength of denture teeth to acrylic resin: effect of thermocycling and polymerisation methods

Background: Failure of bonding between acrylic resin teeth and denture base material is a considerable problem for patients who wear complete dentures. Objective: The purpose of this study was to evaluate the bond strength between acrylic resins and resin denture teeth with different thermocycling and polymerisation methods. Materials and methods: Microwave‐polymerised (Onda‐Cryl), heat‐polymerised (Clássico) and autopolymerising (Jet) acrylic resins and resin denture tooth (Biotone) were used. The acrylic resins were polymerised according to the following: (A) microwave – fast cycle, Onda‐Cryl; (B) microwave – long cycle, Onda‐Cryl; (C) microwave – manufacturer’s cycle, Onda‐Cryl; (T) water bath – long cycle, Clássico; and (Q) bench polymerisation cycle, Jet. Twenty specimens were prepared for each polymerisation method. Ten were thermocycled, and 10 did not receive thermocycling. For the purpose of the study, a shear test was used. Data were analysed with the Kruskal–Wallis test and Dunn’s multiple comparisons test (p = 0.05). Results: Cycles B, C and T were similar (p > 0.05), and had statistically higher bonding values than cycles A and Q (p < 0.05) for the thermocycled and non‐thermocycled groups. For all cycles, there were no statistically significant differences between thermocycled and non‐thermocycled groups (p > 0.05), except for cycle Q (p = 0.0038). Conclusion: Thermocycling decreased the bond strength, but not significantly for microwave and heat‐polymerised cycles. Regarding the bond strength of denture teeth to acrylic resin, the fast microwave polymerisation cycle should be avoided when polymerising Onda‐Cryl acrylic resin. In addition, Jet acrylic resin is not adequate for use in denture repair.     

Treatment protocol for denture stomatitis,prior to anatomical molding

doi: 10.1111/j.1741‐2358.2012.00661.x Treatment protocol for denture stomatitis, prior to anatomical molding Background: Microorganisms of the genus Candida have been recovered from complete dentures made of acrylic resin, with high numbers of colony forming units and species diversity, which can act as infectious agents causing chronic atrophic candidiasis (denture stomatitis). Objective: The objective of this paper is present a treatment protocol for chronic atrophic candidiasis (denture stomatitis). Materials and Methods: The work describes three cases of totally edentulous patients presenting palate stomatitis who were submitted for treatment associating denture rebasing with chemically‐activated acrylic resin, night immersion in 2.5% sodium hypochlorite and use of topical antifungals for two weeks. Results: In all cases, remission of the inflammatory process occurred. Conclusion: The proposed treatment protocol proved to be to be effective.     

Denture disinfection by microwave energy: influence of Candida albicans biofilm

doi: 10.1111/j.1741‐2358.2010.00439.x Denture disinfection by microwave energy: influence of Candida albicans biofilm Objective: This study evaluated the influence of the area of Candida albicans biofilm on denture disinfection by microwave energy. Materials and methods: Candida albicans biofilm was allowed to form for 72 h on resin discs, and three small coverage or seven large coverage discs were placed onto the palatal surface of sterile maxillary dentures. Each denture was immersed in 200 ml distilled water and individually irradiated at a power of 450, 630 or 900 W for different time intervals (1, 2 or 3 min) (n = 6). The effectiveness of disinfection was evaluated by counting the residual cells. The data were analysed by anova and Tukey’s HSD test (α = 0.05). Pearson’s correlation test was performed to determine the correlation between effectiveness of sterilisation and temperature. Results: Dentures with a larger area of biofilm demanded a longer irradiation exposure to achieve disinfection (p < 0.001), irrespective of power setting, and in this time no yeast growth was detected. Dentures with small areas of biofilm were disinfected after 1 min at 900 W and 2 min at 450 or 630 W. A positive correlation was found between water temperature and effectiveness of disinfection (r = 0.6170; p < 0.001). Conclusion: The C. albicans biofilm area influenced disinfection by microwave energy; therefore dentures with larger biofilm areas required longer irradiation exposure to be disinfected.     

Peppermint (Mentha piperita) inhibits microbial biofilms in vitro

Microbial biofilms have become increasingly problematic in the food processing and medical industries where they cause food and surface contamination. Biofilms have also been implicated as the cause of serious infections in humans as their occurrence makes it difficult to treat common infections and the likelihood of recurrent infections is high. Due to emerging resistance, conventional control methods are fast becoming ineffective. In this study, the use of a selection of commercial plant extracts is investigated. The inhibitory effects of eight herbal extracts on the development of microbial biofilms was investigated against clinical and reference strains of Pseudomonas aeruginosa and Candida albicans. The antimicrobial activity was investigated on the planktonic forms using the minimum inhibitory concentration assay. The extracts that showed the highest antimicrobial activity against the two test organisms were Echinacea angustifolia (cone flower), Mentha piperita (peppermint) and Rosmarinus officinalis (rosemary) with minimum inhibitory concentration values between 0.38 and 2.5 mg/ml. The crystal violet assay was used to assess the effect of pre-treating a surface with plant extracts on cell attachment and the extent of biofilm development following exposure to extracts (biofilm biomass). Most of the extracts reduced microbial colonization by at least 50%. In contrast, preformed biofilms were less responsive to the majority of extracts, thus growth inhibition was more difficult to achieve. Mentha piperita was the only extract that showed some antibiofilm activity against both pathogens.     

Non-glucan Attached Proteins of Candida albicans Biofilm Formed on Various Surfaces

Non-glucan attached proteins of the cell surface and extracellular matrix of Candida albicans biofilms formed on two catheter surfaces and denture acrylic were examined. The SDS-PAGE protein profiles of these proteins compared with that obtained from planktonic yeast cells and germ tubes were generally similar. This observation suggested that this class of biofilm surface proteins is not composed of a unique set of extracellular proteins or that one or a few proteins dominate the non-glucan attached proteins of biofilm. However, differences were observed in the proteins obtained from biofilm formed on one catheter surface and two proteins, Grp2p and ORF19.822p, identified by mass spectrometry following two-dimensional separation. These proteins have previously been associated with drug resistance and their presence or abundance appeared to be influenced by the surface on which the biofilm was formed.     

A novel in vitro flat‐bed perfusion biofilm model for determining the potential antimicrobial efficacy of topical wound treatments

Aims: To develop an in vitro flat‐bed perfusion biofilm model that could be used to determine the antimicrobial efficacy of topically applied treatments. Methods and Results: Pseudomonas aeruginosa and Staphylococcus aureus biofilms were grown within continuously perfused cellulose matrices. Enumeration of the biofilm density and eluate was performed at various sampling times, enabling determination of the biofilm growth rate. Two antimicrobial wound dressings were applied to the surface of mature biofilms and periodically sampled. To enable real‐time imaging of biofilm growth and potential antimicrobial kinetics, a bioluminescent Ps. aeruginosa biofilm was monitored using low‐light photometry. Target species produced reproducible steady‐state biofilms at a density of c. 10⁷ per biofilm support matrix, after 24‐h perfusion. Test dressings elicited significant antimicrobial effects, producing differing kill kinetic profiles. There was a good correlation between photon and viable count data. Conclusions: The model enables determination of the antimicrobial profile of topically applied treatments against target species biofilms, accurately differentiating bactericidal from bacteriostatic effects. Moreover, these effects could be monitored in real time using bioluminescence. Significance and Impact of the Study: This is the first in vitro biofilm model which can assess the antimicrobial potential of topical therapies in a dynamic growth environment.