Inhibition of Candida albicans biofilm formation on denture material

doi:10.1016/j.tripleo.2009.01.021

Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology, and Endodontology

Volume 107, Issue 5, May 2009, Pages 669-672

Abstract presented at the American Association for Dental Research Annual session, Orlando, Florida, March 6, 2006.

https://doi.org/10.1016/j.tripleo.2009.01.021 Get rights and content

Objective

The aim was to determine the ability of several thin-film polymer formulations, with and without incorporated antifungals, to inhibit Candida albicans biofilm growth on denture material. The inhibition of C. albicans biofilms on maxillary dentures could play a significant role in preventing the development of denture stomatitis.

Study design

Low-porosity and high-porosity thin-film polymer formulations were used and one of the following fungicides was added: 1) chlorhexidine diacetate at 1.0%; 2) nystatin at 1.0%; or 3) amphotericin B at 0.1%. These coatings were placed on rectangular (12 × 10 mm) dental resin material samples. A subset of the coated dental materials were brushed to simulate denture cleaning for 1 minute per day for 1 year. Candida albicans biofilms were formed on polymethylmethacrylate (PMMA) specimens placed in 24-well polystyrene plates, and the extent of biofilm formation on coated and noncoated specimens was assessed using a 2,3-bis(2-methoxy-4-nitro-5-sulfo-phenyl)-2H-tetrazolium-5-carboxanilide (XTT) reduction assay.

Results

Thin-film polymer PMMA coatings alone, without an antifungal agent, produced a small significant reduction in C. albicans biofilm formation compared with control PMMA. However, incorporation of antifungal medications into the thin-film polymer reduced biofilm formation between 70% and 80% with nystatin, and between 50% and 60% with amphotericin B. Biofilm reduction with chlorhexidine (up to 98%) was significantly greater than all other formulations tested (P < .025).

Conclusion

This novel thin-film coating with various antifungals effectively inhibits C. albicans biofilm formation and should be evaluated as a potential preventive therapy for denture stomatitis.

Section snippets

Materials and Methods

We decided to evaluate 2 variations of the polymer formula by altering porosity. On one hand we felt that the more porous material may provide an improved substrate for biofilm growth. On the other hand, we wanted to determine if higher porosity increased the ability of antifungals to inhibit biofilm growth. The formulations were designated “low porosity” and “high porosity.” Rectangular (12 × 10 mm) dental material (DM) samples were prepared by pouring a mixture of 1 part Dentsply Repair

Results

Fig 1 shows the biofilm inhibition of the thin-film polymer formulas alone after the brushing test was applied. There was >20% inhibition of biofilm formation with the high-porosity film but virtually no inhibition with the low-porosity film. This difference was statistically significant (P < .005).

Fig 2 shows biofilm inhibition of both thin-film polymer formulas with medications after brushing. All 3 medications showed >50% improvement over the thin film alone. However, both chlorhexidine

Discussion

The thin-film polymer appears to inhibit biofilm formation when applied without antifungal medications. Porosity of this formula appears to be significant, with the high-porosity formula showing significant inhibition of biofilm formation. We were surprised at this finding, because we thought that a more porous substrate would promote biofilm formation. However, the thin-film polymer surface even of the high-porosity formula is much less porous than PMMA, so this porosity may be too small to

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Response to fluconazole and itraconazole of Candida spp. in denture stomatitis
Mycoses
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The objective was to synthesize halloysite nanotubes loaded with chlorhexidine (HNT/CHX) and evaluate the antimicrobial activity, microhardness, color change, and surface characteristics of an experimental self-curing acrylic resin containing varying concentrations of the synthesized nanomaterial.
The characterization of HNT/CHX was carried out by calculating incorporation efficiency, morphological and compositional, chemical and thermal evaluations. SAR disks were made containing 0 %, 3 %, 5 %, and 10 % of HNT/CHX. Specimens (n = 3) were immersed in distilled water and spectral measurements were carried out using UV/Vis spectroscopy to evaluate the release of CHX for up to 50 days. The antimicrobial activity of the composite against Candida albicans and Streptococcus mutans was evaluated by disk-diffusion test. Microhardness, color analyses (ΔE), and surface roughness (Ra) (n = 9) were performed before and after 30 days of immersion. Data were analyzed using ANOVA/Bonferroni. {Results.} The incorporation efficiency of CHX into HNT was of 8.15 %. All test groups showed controlled and cumulative CHX release up to 30 or 50 days. Significant antimicrobial activity was verified against both microorganisms (p < 0.001). After the 30-day immersion period, the 10 % HNT/CHX group showed a significant increase in hardness (p < 0.05) and a progressive color change (p < 0.001). At T0, the 5 % and 10 % groups exhibited Ra values similar to the control group (p > 0.05), while at T30, all groups showed similar roughness values (p > 0.05). {Significance.}
The modification of a SAR with HNT/CHX provides antimicrobial effect and controlled release of CHX, however, the immediate surface roughness in the 3 % group was compromised when compared to the control group.
Silver nanoparticles in denture adhesive: An antimicrobial approach against Candida albicans
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To evaluate the antimicrobial potential of silver nanoparticles (Ag NPs) synthesized using three different routes (ultraviolet light, Turkevich, and green chemistry method using Glycine max extract) associated with COREGA® denture powder adhesive.
Heat-cured acrylic resin specimens were treated with different Ag NPs associated with the adhesive (AD + Ag UV, AD + Ag Turk, and AD + Ag Gm groups). As controls, the specimens were treated with a combination of adhesive and nystatin (AD + Nyst group), only adhesive (AD group), or submerged on the surface of the specimens (PBS group). After the treatments, biofilms of C. albicans developed for 3, 6, and 12 h on the specimen surfaces. The biofilm was quantified using colony-forming units per milliliter, colorimetric assay, and confocal laser scanning microscopy.
Regardless of the period, we observed an inhibition of fungal load and a reduction in metabolic activity and biofilm mass in the resin specimens treated with the combinations AD/Ag NPs, compared to AD and PBS. The antimicrobial action of the AD + Turk and AD + Ag Gm groups was similar than that for the AD + Nyst group in all periods and viability tests, except for the biofilm mass (12 h).
The COREGA® adhesive with Ag NPs, mainly those synthesized using the Turkevich and Glycine max methods, showed excellent antimicrobial activity against C. albicans biofilms, maintained for up to 12 h.
The association of Ag NPs to the adhesive can add preventive or therapeutic effects against denture stomatitis, to this prosthetic material.
Development of a chlorhexidine delivery system based on dental reline acrylic resins
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The high recurrence rate of common denture stomatitis after antifungal treatment is still concerning. This condition is caused by low patient compliance and incomplete local elimination of the main etiological factor — Candida albicans, often associated with other microorganisms, such as Streptococcus species. Impregnating denture materials with antimicrobials for local delivery is a strategy that can overcome the side effects and improve the efficacy of conventional treatments (topical and/or systemic). In this work, we describe the development of three hard autopolymerizing reline acrylic resins (Kooliner, Ufi Gel Hard, and Probase Cold) loaded with different percentages of chlorhexidine (CHX). The novel formulations were characterized based on their antimicrobial activity, mechanical, morphological and surface properties, in-vitro drug release profiles, and cytotoxicity. The addition of CHX in all resins did not change their chemical and mechanical structure. Among all the tested formulations, Probase Cold loaded with 5 wt% CHX showed the most promising results in terms of antimicrobial activity and lack of serious detrimental mechanical, morphological, surface, and biological properties.
Determining growth inhibition of Candida albicans biofilm on denture materials after application of an organoselenium-containing dental sealant
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Denture stomatitis is a chronic inflammatory condition caused by the formation of Candida albicans biofilm on denture bases. It is associated with aggravating intraoral pain, itching, and burning sensations. It can also potentiate cardiovascular diseases and aspiration pneumonia. The problem has thus far eluded efficient, toxic-free, and cost-effective solutions.
The purpose of this in vitro study was to investigate the effectiveness of organoselenium to inhibit the formation of C. albicans biofilm on the surface of acrylic resin denture base materials when it is either incorporated into the acrylic resin material or coated on the denture surface as a light-polymerized surface sealant.
Sixty heat-polymerized polymethyl methacrylate disks were fabricated and assigned to 4 groups (n=15): disks coated with a light-polymerized organoselenium-containing enamel surface sealant (DenteShield), disks impregnated with 0.5% organoselenium (0.5% selenium), disks impregnated with 1% organoselenium (1% selenium), and disks without organoselenium (control). C. albicans biofilm was grown on each disk which had been placed in a well of the microtiter plate containing 1-mL brain heart infusion broth inoculated with C. albicans. The plates were incubated aerobically at 37 °C for 48 hours. A confocal laser scanning microscope was used to determine the biofilm thickness, biomass, and live/dead cell ratio. Biofilm morphology was examined with scanning electron microscopy, whereas microbial viability was quantified by the spread plate method. The data were analyzed by using ANOVA and Tukey-Kramer multiple comparisons (α=.05).
The microbial viability, biofilm thickness, biofilm biomass, and live/dead cell ratio were lower (P<.001) on disks in the test groups (DenteShield, 0.5% selenium, 1% selenium) when compared with the control group, with these variables being lowest in the 0.5% selenium and 1% selenium groups. The 0.5% selenium and 1% selenium groups did not differ significantly from each other in any of the variables (P>.05). Scanning electron microscope images showed inhibition of both biofilm growth and yeast to hyphae transition in the DenteShield, 0.5% selenium, and 1% selenium groups, with visible disruption of the biofilm morphology.
The present study demonstrated that organoselenium, whether incorporated into or coated on the surface of an acrylic resin denture base material, has the potential to inhibit Candida albicans biofilm growth on denture surfaces and as such can be clinically useful for the prevention of denture stomatitis.
Fracture strength of Er,Yag laser treated PMMA denture-based polymer (DBP) colonized with C. albicans, S. aureus, S.mutans, and E.coli
2022, Photodiagnosis and Photodynamic Therapy
Citation Excerpt :
Silva and their colleagues found 2% CHX to be effective in disinfecting PMMA denture bases. Also, Redding et al. deduced analogous results [44]. However, the effect of CHX is dependent on the pH of the environment and the manifestation of organic matter [15].
To assess and equate the efficacy of different disinfection protocols autoclave, chlorhexidine (CHX), PDT utilizing Rose Bengal (RB), chitosan, and Er, Yag laser and their effect on fracture strength of PMMA denture-based polymer (DBP) colonized with C. Albicans, S. aureus, S.mutans, and E.coli.
A total of 50 (n = 10) PMMA DBP were manufactured and adulterated with the American Type Culture Collection (ATCC) of diverse microbial colonies inhabited by C. Albicans, S. aureus, S.mutans, and E.coli. The specimens were subjected to different denture disinfection approaches by randomly distributing in into five groups i.e., Er, Yag laser, RB, autoclave, CHX, and Chitosan, respectively for appraising antimicrobial effectiveness. PMMA fracture load was also assessed and statistical analysis was performed for CFU/mL (log10) of exposed C. Albicans, S. aureus, S.mutans, and E.coli by one-way ANOVA and Tukey's multiple comparison test at a significance level of p < 0.05.
Intergroup comparison disclosed that denture disinfection with Er, Yag laser, autoclave, Chitosan, and CHX (control) validated comparable antimicrobial efficacy to denture against all inspected CFU/mL (log10) (p>0.05). The intragroup comparison revealed that DBP sanitization with Er, Yag laser, autoclave, Chitosan, RB, and CHX substantiated equivalent effective antimicrobial efficacy in plummeting CFU/mL (log10) of S. mutans and E. coli (p>0.05) but in consideration to S.aureus and C.albicans, all groups resulted in declining their count except 5µm RB activated by PDT(p < 0.05). No significant difference was perceived in fracture load of PMMA denture base among Er, Yag laser, RB, chitosan, and CHX (control) (p > 0.05) except autoclave decontamination procedure that indicated the least fracture strength of DBP when disinfected (p < 0.05).
Er, Yag laser, and Chitosan activated by PDT have the potential to be used as an alternative to chlorhexidine for disinfecting Polymethyl methacrylate denture base as they demonstrated the highest antimicrobial efficacy against E. coli, C. Albicans, S aureus, and S. mutans with optimal fracture load.
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Specimens of 3D printing denture base polymers were prepared by two printers, namely, stereolithography (SLA, Form 3B) and digital light processing technology (DLP, Solflex 350 plus). Three build angles were used: 0°, 45°, and 90°. Surface topography was examined by scanning electron microscopy. Also, arithmetical mean height (S_a) values were calculated. An adhesion test was performed to observe initial C. albicans adhesion to the specimens. The data were statistically analyzed using the two-way analysis of variance and Tukey's multiple comparison test.
The data of S_a values had statistically significant differences, which were mainly determined by the main factor of build angle (p < 0.05). Moreover, the quantitative results of C. albicans adhesion exhibited no significant differences: printing techniques (p = 0.7794) and build angle (p = 0.0589), respectively.
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: Supported by an International Association for Dental Research/Glaxo Smith Kline Innovation in Oral Care Award in 2004 to the University of Texas Health Science Center at San Antonio and Biomedical Development Corporation.

¹: B. Bhatt is a former employee and G. Siegel is a current employee and officer of Biomedical Development Corporation

²: S. Redding and R. Rawls have participated as investigators on several Small Business Innovation Research grants to Biomedical Development Corporation.

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Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology, and Endodontology

Objective

Study design

Results

Conclusion

Section snippets

Materials and Methods

Results

Discussion

References (12)

Antifungal therapy in oropharyngeal mycotic infections

Oral Surg Oral Med Oral Pathol Oral Radiol Endod

Denture stomatitis: a role for Candida biofilms

Oral Surg Oral Med Oral Pathol Oral Radiol Endod

Denture stomatitis IThe etiology in relation to trauma and infection

Acta Odontol Scand

Denture Stomatitis: a review

J Oral Rehab

Etiology, pathogenesis, therapy, and prophylaxis of oral yeast infections

Acta Odontol Scand

Response to fluconazole and itraconazole of Candida spp. in denture stomatitis

Mycoses

Cited by (113)

Microbiological, physicomechanical, and surface evaluation of an experimental self-curing acrylic resin containing halloysite nanotubes doped with chlorhexidine

Silver nanoparticles in denture adhesive: An antimicrobial approach against Candida albicans

Development of a chlorhexidine delivery system based on dental reline acrylic resins

Determining growth inhibition of Candida albicans biofilm on denture materials after application of an organoselenium-containing dental sealant

Fracture strength of Er,Yag laser treated PMMA denture-based polymer (DBP) colonized with C. albicans, S. aureus, S.mutans, and E.coli

Effect of additive manufacturing method and build angle on surface characteristics and Candida albicans adhesion to 3D printed denture base polymers

Oral medicineInhibition of Candida albicans biofilm formation on denture material

Objective

Study design

Results

Conclusion

Section snippets

Materials and Methods

Results

Discussion

Oral Surg Oral Med Oral Pathol Oral Radiol Endod

Oral Surg Oral Med Oral Pathol Oral Radiol Endod

Denture stomatitis IThe etiology in relation to trauma and infection

Acta Odontol Scand

Denture Stomatitis: a review

J Oral Rehab

Etiology, pathogenesis, therapy, and prophylaxis of oral yeast infections

Acta Odontol Scand

Response to fluconazole and itraconazole of Candida spp. in denture stomatitis

Mycoses

Oral medicine
Inhibition of Candida albicans biofilm formation on denture material