Lisa Coulthwaite

Potential pathogenic aspects of denture plaque.

Abstract Oral health status declines with age and as a result the need for removable prostheses increases. Oral health is a reflection of one’s general health, affecting the ability of an individual to eat and speak, and contributes significantly to a sense of confidence and well-being. Currently, there are 15 million denture wearers in the UK, representing a significant consumer base and a special healthcare consideration. The microbiology of denture plaque has received little attention in comparison with dental plaque, yet it differs in location and composition. Denture plaque and poor denture hygiene is associated with stomatitis (Candida infection), may also serve as a reservoir of potentially infectious pathogens, and may contribute to oral malodour and to caries and periodontitis in people who have remaining natural teeth. Oral bacteria have been implicated in bacterial endocarditis, aspiration pneumonia, gastrointestinal infection and chronic obstructive pulmonary disease, among others, and dentures offer a reservoir for microorganisms associated with these infections. An effective oral hygiene regimen is important to control denture plaque biofilm and contributes to the control of associated oral and systemic diseases.

The Microbiological Origin of Fluorescence Observed in Plaque on Dentures during QLF Analysis

The aim of this study was to determine the microbiological origin of plaque fluorescence observed during quantitative light-induced fluorescence (QLF) analysis. Plaque was sampled from dentures, because of easy accessibility and the homogeneous background provided by the denture tooth during imaging, and the acknowledged comparability to occlusal plaque. Forty removable poly(methyl methacrylate) dentures were screened for the presence of fluorescent plaque deposits during QLF analysis. Dentures were photographed, QLF images were recorded and samples of fluorescent plaque were taken. Plaque samples were cultured on fastidious anaerobe agar, Wilkins Chalgren agar and Sabourauds dextrose agar. Plates were screened under QLF and fluorescent colonies were subcultured and identified. Areas of red, orange and green fluorescence were detected on the fitting and non-fitting surfaces of dentures. The red and orange fluorescing species were Prevotella melaninogenica, Actinomyces israelii and Candida albicans, which are generally acknowledged to be secondary colonisers, present in more mature plaque. Green fluorescence was observed in streptococcal species (early colonisers) and Fusobacterium nucleatum (important organism in plaque development). Non-fluorescent colonies were also cultured. Plaque which accumulates on susceptible surfaces tends to be associated with caries, but it may be its maturity, rather than the presence of cariogenic streptococci, that is more likely to provide a microbiological link between red fluorescence and caries.

Biofilm development by blastospores and hyphae of Candida albicans on abraded denture acrylic resin surfaces

STATEMENT OF PROBLEM Candida albicans is a known etiologic agent of denture stomatitis. Candida hyphae exhibit the ability to respond directionally to environmental stimuli. This characteristic is thought to be important in the penetration of substrata such as resilient denture liners and host epithelium. It has been suggested that hyphal production also enhances adhesion and survival of Candida on host and denture surfaces. Surface roughness, in addition, can enhance adhesion where stronger interactions occur between cells and surface features of similar dimensions. PURPOSE The purpose of this study was to assess the development of hyphal and blastospore biofilms on abraded denture acrylic resin specimens and measure the ease of removal of these biofilms. MATERIAL AND METHODS Biofilms were grown for 48 hours on abraded 1-cm² denture acrylic resin specimens from adhered hyphal phase C albicans or from adhered blastospores. Subsequently, all specimens were stained with Calcofluor White and examined with confocal scanning laser microscopy. Biofilms were removed by vortex mixing in sterile phosphate buffered saline solution. Removed cells were filtered (0.2-μm pore size). Filters were dried at 37°C for 24 hours for dry weight measurements. Any cells that remained on the acrylic resin specimens were stained with 0.03% acridine orange and examined with epifluorescence microscopy. RESULTS Biofilms grown from both cell types contained all morphologic forms of C albicans. Although the underlying surface topography did not affect the amount of biofilm produced, biofilms grown from hyphal phase Candida were visibly thicker and had greater biomass (P<.05). These biofilms were less easily removed from the denture acrylic resin, especially in the case of rougher surfaces, evidenced by the higher numbers of retained cells (P≤.05). CONCLUSION The presence of hyphae in early Candida biofilms increased biofilm mass and resistance to removal. Increased surface roughness enhances retention of hyphae and yeast cells, and, therefore, will facilitate plaque regrowth. Therefore, minimization of denture abrasion during cleaning is desirable.

The effect of dentifrice abrasion on denture topography and the subsequent retention of microorganisms on abraded surfaces.

STATEMENT OF PROBLEM Denture surfaces provide hard nonshedding niches for the adhesion and subsequent accumulation of oral microorganisms into denture plaque, which can harbor various potential pathogens linked with oral mucosal lesions and inhalation pneumonia. The initial adhesion is the prerequisite for subsequent biofilm growth, and surface roughness niches facilitate this process by trapping cells. Retained microorganisms are then able to proliferate when the denture is returned to the oral cavity. PURPOSE The purpose of this study was to measure the amount and strength of the attachment of microorganisms to a roughened denture acrylic resin surface. An increase in surface roughness increases the retention of microorganisms and a greater amount of cell-surface contact interface may increase the strength of adhesion and, therefore, retention. Cleaning denture surfaces with brushes and dentifrices can influence the denture surface topography and, therefore, may affect retention. MATERIAL AND METHODS Denture acrylic resin specimens were abraded to provide different surface roughness. The amount of attachment of Streptococcus oralis or Candida albicans to these surfaces was assessed by measuring the area of a microscopic field covered by stained cells after 1 hour of incubation. The strength of attachment was assessed with atomic force microscopy, whereby an increasing force was applied to the attached cells until they detached from the surface. RESULTS Both bacteria and yeast cells were retained in increasing amounts on surfaces of increasing roughness. Cells were most strongly attached on surfaces whose linear features (scratches) were of comparable size with the cells (the streptococci on the low-abraded surfaces, and the yeast on high-abraded surfaces). CONCLUSION Analysis of findings reveal that even small abrasions may enhance retention on denture surfaces and reduce surface cleanability. The strength of attachment instead of the amount is more important in terms of surface hygiene.

QLF is not readily suitable for in vivo denture plaque assessment.

OBJECTIVES Current methods available for denture plaque assessment utilise visual and planimetric techniques. This paper evaluates the use of the Quantitative Light-induced Fluorescence system (QLF) in image capture of denture plaque and the suitability of these images for planimetric plaque measurement. It is proposed that fluorescence imaging could provide a valuable and sensitive standardising method for plaque assessment in clinical trials for denture cleansing products and denture hygiene. Indeed, the detection of red fluorescent plaque using the QLF system is indicative of black-pigmented obligate anaerobes and mature plaque. METHODS The QLF system was evaluated in a clinical study for use in denture plaque assessment in comparison to white light based image capture. RESULTS Despite appearing as a promising system for denture plaque quantification, this study revealed numerous problems associated with the QLF system including small focal depth, thus large numbers of images and processing time were required. In addition, differential fluorescence of acrylic made images unsuitable for plaque quantification. CONCLUSION QLF is unsuitable for in vivo denture plaque assessment. However, the visualisation of red autofluorescence, indicating mature plaque, remains an important clinical use of QLF for denture hygiene assessment.

The effect of a commercial probiotic drink containing Lactobacillus casei strain Shirota on oral health in healthy dentate people

Background In the past decade, the use of probiotic-containing products has been explored as a potential alternative in oral health therapy. A widely available probiotic drink, Yakult, was evaluated for oral health applications in this longitudinal study. Selected oral health parameters, such as levels and composition of salivary and tongue plaque microbiota and of malodorous gases, in dentate healthy individuals were investigated for changes. The persistence of the probiotic strain in the oral cavity was monitored throughout the study period. Methods A three-phase study (7 weeks) was designed to investigate simultaneously the effect of 4-week consumption of the probiotic-containing milk drink Yakult on the microbiota of saliva and dorsum tongue coating in healthy dentate people (n = 22) and levels of volatile sulphur compounds (VSCs) in morning breath. Study phases comprised one baseline visit, at which ‘control’ levels of oral parameters were obtained prior to the probiotic product consumption; a 4-week period of daily consumption of one 65 ml bottle of Yakult, each bottle containing a minimum of 6.5×109 viable cells of Lactobacillus casei strain Shirota (LcS); and a 2-week washout period. The microbial viability and composition of saliva and tongue dorsum coating were assessed using a range of solid media. The presence of LcS in the oral cavity was investigated using a novel selective medium, ‘LcS Select’. Portable sulphur monitors Halimeter® and OralChromaTM were used to measure levels of VSCs in morning breath. Results Utilization of the LcS Select medium revealed a significant (p < 0.05) but temporary and consumption-dependent presence of LcS in saliva and tongue plaque samples from healthy dentate individuals (n = 19) during the probiotic intervention phase. LcS was undetectable with culture after 2 weeks of ceasing its consumption. Morning breath scores measured with Halimeter and OralChroma were not significantly affected throughout the trial, except in a small number of individual cases where Halimeter scores were significantly reduced during the probiotic intervention period. Natural fluctuations in resident acidogenic populations, and numbers of Candida and anaerobic species, including malodourous Gram-negative anaerobes, were unaffected. Conclusion While no broad ecological changes in the mouth were induced by consumption of Yakult in healthy dentate individuals, findings of this study confirm the temporary and intake-dependent presence of LcS. Future studies could focus on subjects at greater risk of oral infection, where ill-defined microbiota (e.g. an increased presence of periopathogens) or clinically diagnosed halitosis might be significantly affected by consumption of this probiotic.

The effect of a commercial probiotic drink on oral microbiota in healthy complete denture wearers

Background : It is acknowledged that oral and general health status declines with age. The global population of denture wearers is increasing, so is the incidence of denture biofilm-related problems, such as denture-associated stomatitis, aspiration pneumonia and malodour. It has been suggested that consumption of probiotic bacteria may improve oral health. However, the effects of probiotics on the oral microbiota of denture wearers have received little attention. Methods : The aim of this study was to investigate the effect of consumption of a commercial probiotic product (Yakult) on microbiota of saliva, tongue and denture biofilm in healthy complete denture wearers. Eight healthy complete denture-wearing National Health Service (NHS) patients undertook a 7-week trial comprising three phases: baseline; 4-week consumption of one bottle of Yakult per day, each containing a minimum 6.5×109 viable cells of Lactobacillus casei strain Shirota (LcS); 4-week washout period. The microbial viability and composition of saliva, tongue dorsum coating and denture biofilm were assessed using a range of solid selective and indicator media. Questionnaires were used to explore participants’ denture cleaning habits and impact of wearing dentures on their life quality and well-being [modified oral health impact profile (OHIP-14)] prior to and after the study. Results : Seven volunteers (1 male, 6 females) completed the trial. LcS temporarily colonised oral cavity and denture surface. There was no significant change in the viability of Streptococcus mutans, acidogenic microorganisms, total anaerobic species and Gram-negative obligate anaerobes between study phases. There was no obvious effect of LcS on occurrence and viability of Candida. Participants presented a good general knowledge of denture hygiene and their responses to OHIP-14 questionnaires improved after completing the study (p=0.16). Conclusion : It appeared that 4-week consumption of probiotic drink had no overall effect on selected oral parameters in healthy denture wearers despite temporary presence of LcS.