Tatiana Pereira-Cenci

Development of Candida-associated denture stomatitis: new insights

Despite therapeutic progress, opportunistic oral fungal infectious diseases have increased in prevalence, especially in denture wearers. The combination of entrapment of yeast cells in irregularities in denture-base and denture-relining materials, poor oral hygiene and several systemic factors is the most probable cause for the onset of this infectious disease. Hence colonization and growth on prostheses by Candida species are of clinical importance. The purpose of this review is to critically discuss several key factors controlling the adhesion of Candida species which are relevant to denture-associated stomatitis. Although there is some consensus on the role of surface properties, studies on several other factors, as the use of denture liners, salivary properties and yeast-bacterial interactions, have shown contradictory findings. A comprehensive fundamental understanding is hampered by conflicting findings due to the large variations in experimental protocols, while other factors have never been thoroughly studied. Surface free energy and surface roughness control the initial adherence, but temporal changes have not been reported. Neither have in vivo studies shown if the substratum type is critical in dictating biofilm accumulation during longer periods in the oral environment. The contribution of saliva is unclear due to factors like variations in its collection and handling. Initial findings have disclosed that also bacteria are crucial for the successful establishment of Candida in biofilms, but the clinical significance of this observation is yet to be confirmed. In conclusion, there is a need to standardize experimental procedures, to bridge the gap between laboratory and in vivo methodologies and findings and – in general – to thoroughly investigate the factors that modulate the initial attachment and subsequent colonization of denture-base materials and the oral mucosa of patients subjected to Candida infections. Information on how these factors can be controlled is required and this may help to prevent the disease. The societal impact of such information is significant given the magnitude of the candidosis problem worldwide.

The effect of Streptococcus mutans and Candida glabrata on Candida albicans biofilms formed on different surfaces

Although Candida containing biofilms contribute to the development of oral candidosis, the characteristics of multi-species Candida biofilms and how oral bacteria modulate these biofilms is poorly understood. The aim of this study was to investigate interactions between Candida albicans and either Candida glabrata or Streptococcus mutans in biofilms grown on various surfaces, with or without saliva. Hydroxyapatite (HA), polymethylmetacrylate (PMMA) and soft denture liner (SL) discs were used as substratum. Counts of viable micro-organisms in the accumulating biofilm layer were determined and converted to colony forming units per unit surface area. Confocal laser scanning microscopy was used to characterize biofilms and to quantitate the number of hyphae in each condition tested. Viable counts of C. albicans and C. glabrata per mm(2) decreased in the order HA>PMMA>SL (p<0.05). Biofilms grown on saliva-coated specimens harboured fewer C. glabrata than uncoated specimens (p<0.05). Glucose and the presence of S. mutans suppressed C. albicans hyphal formation. Dual Candida species biofilms did not show competitive interaction between the two species. We conclude that Candida biofilms are significantly affected by saliva, substratum type and by the presence of other micro-organisms.

Molecular and Cellular Mechanisms That Lead to Candida Biofilm Formation

Fungal infections in the oral cavity are mainly caused by C. albicans, but other Candida species are also frequently identified. They are increasing in prevalence, especially in denture-wearers and aging people, and may lead to invasive infections, which have a high mortality rate. Attachment to mucosal tissues and to abiotic surfaces and the formation of biofilms are crucial steps for Candida survival and proliferation in the oral cavity. Candida species possess a wide arsenal of glycoproteins located at the exterior side of the cell wall, many of which play a determining role in these steps. In addition, C. albicans secretes signaling molecules that inhibit the yeast-to-hypha transition and biofilm formation. In vivo, Candida species are members of mixed biofilms, and subject to various antagonistic and synergistic interactions, which are beginning to be explored. We believe that these new insights will allow for more efficacious treatments of fungal oral infections. For example, the use of signaling molecules that inhibit biofilm formation should be considered. In addition, cell-wall biosynthetic enzymes, wall cross-linking enzymes, and wall proteins, which include adhesins, proteins involved in biofilm formation, fungal-bacterial interactions, and competition for surface colonization sites, offer a wide range of potential targets for therapeutic intervention.

Relationship between Gap Size and Dentine Secondary Caries Formation Assessed in a Microcosm Biofilm Model

Secondary caries can develop at the tooth-restoration interface, depending on the presence of a gap and its size, but this process could be inhibited by fluoride. The aim of this study was to assess the relationship between gap size and dentine secondary caries adjacent to composite resin (CR) or glass ionomer (GI) restorations, using a microcosm biofilm model in a constant depth film fermentor (CDFF). Dentine discs restored with CR (Z250) or GI (Vitremer) with gap sizes of 0, 50, 100, 180 or 250 μm were mounted on the CDFF. Microcosm biofilms were formed on the restored discs and daily subjected to 8 pulses of 10% sucrose solution. On the 18th day, dentine mineral loss and lesion depth around the restorations were determined by transverse microradiography.The effect of gap size was overall not statistically significant either with regard to mineral loss (p = 0.449) or lesion depth (p = 0.328), but greater mineral loss and lesion depth were found adjacent to CR than to GI (p < 0.001). However, Spearman correlation showed that mineral loss and lesion depth increased with gap size for CR (p < 0.001) but not for GI (p > 0.05). The findings support the conclusion that fluoride released from GI inhibits dentine demineralization adjacent to restorations, irrespective of gap width.

The role of resin cement on bond strength of glass-fiber posts luted into root canals: a systematic review and meta-analysis of in vitro studies.

Because there are several ways to cement glass-fiber posts (GFPs) into root canals, there is no consensus on the best strategy to achieve high bond strengths. A systematic review was conducted to determine if there is difference in bond strength to dentin between regular and self-adhesive resin cements and to verify the influence of several variables on the retention of GFPs. This report followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement. In vitro studies that investigated the bond strength of GFPs luted with self-adhesive and regular resin cements were selected. Searches were carried out in the PubMed and Scopus databases. No publication year or language limit was used, and the last search was done in October 2012. A global comparison was performed between self-adhesive and regular resin cements. Two subgroup analyses were performed: 1) Self-adhesive × Regular resin cement + Etch-and-rinse adhesive and 2) Self-adhesive × Regular resin cement + Self-etch adhesive. The analyses were carried out using fixed-effect and random-effects models. The results showed heterogeneity in all comparisons, and higher bond strength to dentin was identified for self-adhesive cements. Although the articles included in this meta-analysis showed high heterogeneity and high risk of bias, the in vitro literature seems to suggest that use of self-adhesive resin cement could improve the retention of GFPs into root canals.

Effect of Microleakage and Fluoride on Enamel-Dentine Demineralization around Restorations

There is no consensus about an association between microleakage and secondary caries, especially considering the presence of fluoride (F) at the tooth/restoration interface. Thus, a randomized, double-blind, crossover study was carried out to evaluate in situ the effect of microleakage on caries around enamel-dentine restorations in the presence of F from dental materials or dentifrice, either alone or in combination. In 4 phases of 14 days each, 14 volunteers wore palatal devices containing dental slabs restored with composite resin (CR) or resin-modified glass ionomer cement (GI). Restorations were made without leakage (L–), following the recommended adhesive procedures, or with leakage (L+), in the absence of adhesive procedures. Plaque-like biofilm (PLB) was left to accumulate on the restored slabs, which were exposed extraorally to a 20% sucrose solution 10×/day. The volunteers used a non-F (NF) or an F (FD) dentifrice 3×/day, depending on the experimental phase. No differences were found between L+ or L– restorations (p > 0.05). Higher demineralization in both enamel and dentine around CR restorations was observed under NF (p < 0.05). F concentration was higher in the fluid of PLB exposed to FD or formed onto GI restoration (p < 0.05). These results suggest that while microleakage does not affect caries development, GI or FD may maintain increased F levels in the PLB, thereby decreasing caries progression.

Efficacy of denture cleansers on Candida spp. biofilm formed on polyamide and polymethyl methacrylate resins

STATEMENT OF PROBLEM Although new materials have emerged as options to fabricate removable dental prostheses, the development of Candida biofilms on these materials and the effectiveness of methods to control these pathogenic biofilms are poorly understood. PURPOSE The purpose of this study was to evaluate the efficacy of denture cleansers on Candida single- and dual-species biofilms formed on polyamide resin. MATERIAL AND METHODS Polymethyl methacrylate (PMMA) resin (Acron MC) and polyamide resin (Flexite M.P.) specimens (n=116) were prepared, and their surface roughness was standardized (0.34 ±0.02 μm). Surface free energy (SFE) was measured for some specimens (n=20 per resin), while the remainder were randomly divided by lottery into 24 groups (n=8) for biofilm assay. C. albicans and/or C. glabrata biofilm was formed for 72 hours, and then specimens were treated with an enzymatic cleanser solution (Polident 3 Minutes), a cleanser solution (Corega Tabs), or 0.5% sodium hypochlorite (NaOCl) solution. Water served as the negative control. Remaining adherent microorganisms were removed from the treated specimens by ultrasonic waves, and colony-forming units (CFU) of each microorganism were calculated. SFE data were analyzed by 1-way ANOVA, and Candida species data were analyzed by 3-way ANOVA followed by the Tukey-Kramer test (P=.05). RESULTS All tested biofilms displayed significantly higher growth on polyamide resin (P<.001), which presented the lowest SFE. Denture cleansers significantly decreased Candida levels; however, the 0.5% NaOCl solution was the only effective cleanser. C. glabrata revealed significantly higher CFU counts under all experimental conditions (P<.001). CONCLUSIONS The highest Candida spp. biofilm growth was shown to occur on polyamide resin when compared with PMMA. Denture cleansers were able to remove Candida spp. biofilm formed on both denture base resins.

MMP Inhibitors on Dentin Stability A Systematic Review and Meta-analysis

The aim of this study was to systematically review the literature for in vitro and ex vivo studies that evaluated the effect of matrix metalloproteinase (MMP) inhibitors during the adhesive procedure on the immediate and long-term resin-dentin bond strength. The search was conducted in 6 databases with no publication year or language limits, following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement. From 1,336 potentially eligible studies, 48 were selected for full-text analysis, and 30 were included for review, with 17 considered in the meta-analysis. Two reviewers independently selected the studies, extracted the data, and assessed the risk of bias. Pooled effect estimates were expressed as the weighted mean difference between groups. The most used MMP inhibitor was chlorhexidine (CHX). Immediate bond strength results showed no difference between 2% CHX and control; however, a difference was found between 0.2% CHX and control at baseline. After aging, CHX presented higher bond strength values compared to control groups (p < .05). However, this was not observed for longer periods of aging. High heterogeneity was found in some comparisons, especially for the water storage aging subgroup. Subgroup analyses showed that self-etching and etch-and-rinse adhesives are benefited by the CHX use. From the studies included, only 1 presented low risk of bias, while the others showed medium or high risk of bias. The use of MMP inhibitors did not affect the immediate bond strength overall, while it influenced the aged bond strength. Aging procedures influenced bond strength values of the dentin adhesion stability.

Influence of thermal stress on marginal integrity of restorative materials

The aim of this study was to evaluate the influence of thermal stress on the marginal integrity of restorative materials with different adhesive and thermal properties. Three hundred and sixty Class V cavities were prepared in buccal and lingual surfaces of 180 bovine incisors. Cervical and incisal walls were located in dentin and enamel, respectively. Specimens were restored with resin composite (RC); glass ionomer (GI) or amalgam (AM), and randomly assigned to 18 groups (n=20) according to the material, number of cycles (500 or 1,000 cycles) and dwell time (30 s or 60 s). Dry and wet specimens served as controls Specimens were immersed in 1% basic fuchsine solution (24 h), sectioned, and microleakage was evaluated under x40 magnification. Data were analyzed by Kruskal-Wallis and Mann-Whitney tests: Thermal cycling regimens increased leakage in all AM restorations (p<0.05) and its effect on RC and GI restorations was only significant when a 60-s dwell time was used (p<0.05). Marginal integrity was more affected in AM restorations under thermal cycling stress, whereas RC and GI ionomer restoration margins were only significantly affected only under longer dwell times.

The Effect of Polishing Techniques and Time on the Surface Characteristics and Sealing Ability of Resin Composite Restorations After One-year Storage

Since there is a lack of information on the surface properties of composite restorations achieved with standard polishing procedures after aging processes, this study evaluated the effects of immediate (IM) and delayed (DE) polishing on the surface roughness (Ra), microhardness (KHN) and microleakage (ML) of microfilled (Filtek A110) and hybrid (Filtek Z250) resin composites after one-year storage. Standardized preparations were made on the buccal surface of 256 bovine teeth, where half were restored with each composite. For each composite, the specimens were randomly allocated to two subgroups. The first group (IM) was polished immediately after gross finishing using three different systems/techniques (n=16): Sof-Lex, Flexicups and Flexicups + Jiffy Polishing Brush + Flexibuffs (sequential), then stored for three weeks in saline. The DE group was stored for two weeks, polished with the same systems and stored for one week. From each subgroup, eight specimens were assessed after three weeks regarding Ra, KHN and ML (baseline), andthe eight remaining specimens were stored for one year before analysis. The data were analyzed (alpha=0.05) with ANOVA, paired Students t-test (Ra and KHN) or Kruskal-Wallis and Signed Rank tests (microleakage). After one year, microfilled resin composite specimens showed the lowest Ra and KHN (p<0.05). N o differencein microleakage was observed among the different groups (p>0.05). The sequential technique provided the lowest roughness and Sof-Lex the lowest hardness (p<0.05). IM showed similar or better performance than DE for ML and Ra (p<0.05). In conclusion, aging increased the composites Ra and ML in all experimental conditions (p<0.05).