Melbourne Dental School - Research Publications

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    Taxonomy of Oral Bacteria
    Byrne, SJ ; Butler, CA ; Reynolds, EC ; Dashper, SG ; Gurtler, V ; Trevors, JT (ELSEVIER ACADEMIC PRESS INC, 2018-01-01)
    The oral cavity is a collection of diverse microenvironments, each inhabited by a community of microorganisms, the majority of which are bacteria and their phages. Given the appropriate conditions, some of these bacteria can cause destruction of the teeth or their supporting hard and soft tissues. For over 300 years microbiologists have been characterising these microbial communities, in both oral health and disease. In this chapter, we take the reader on a journey through time as we discuss the various methods that have been utilised in the characterisation of the bacteria calling the oral cavity home, and how the use of these methods has informed our understanding of oral bacterial communities and the diversity of their members.
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    The prebiotic effect of CPP-ACP sugar-free chewing gum
    Fernando, JR ; Butler, CA ; Adams, GG ; Mitchell, HL ; Dashper, SG ; Escobar, K ; Hoffmann, B ; Shen, P ; Walker, GD ; Yuan, Y ; Reynolds, C ; Reynolds, EC (ELSEVIER SCI LTD, 2019-12)
    OBJECTIVES: To determine if chewing gum containing casein phosphopeptide stabilised amorphous calcium phosphate (CPP-ACP) promoted an increase in the abundance of Streptococcus sanguinis and other species associated with dental health in supragingival plaque in a clinical study. MATERIALS AND METHODS: Nineteen participants were recruited for a three-leg cross-over, randomised, controlled clinical trial. Participants chewed a sugar-free gum with or without CPP-ACP six times daily for 20 min over two weeks. The study also involved no gum chewing (no gum) for the same two week period. Participants were randomly assigned to one of the test gums or no gum for each intervention period. Participants abstained from oral hygiene and had washout periods of two weeks between intervention periods. After each intervention period, supragingival plaque was collected and analysed for bacterial composition by sequencing the V4 variable region of the 16S rRNA gene. Data were analysed using a linear mixed model. RESULTS: The CPP-ACP gum intervention produced a significant (p < 0.01) increase in the proportions of S. sanguinis (112%), as well as the commensal species Rothia dentocariosa (127%), Corynebacterium durum (80%) and Streptococcus mitis (55%) when compared with the no gum intervention. All the species that were promoted by the CPP-ACP gum are known to possess one or both of the alkali-producing enzymes arginine deiminase and nitrate reductase. CONCLUSION: This clinical study demonstrated that chewing a sugar-free gum containing CPP-ACP promoted prebiosis by significantly increasing the proportion of S. sanguinis and other health-associated bacterial species in supragingival plaque. CLINICAL SIGNIFICANCE: Regular chewing of CPP-ACP sugar-free gum increases the proportions of health-associated commensal species in supragingival plaque to promote prebiosis and oral homeostasis.
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    Polymicrobial interactions of Candida albicans and its role in oral carcinogenesis
    Arzmi, MH ; Dashper, S ; McCullough, M (WILEY, 2019-08)
    The oral microbiome is composed of microorganisms residing in the oral cavity, which are critical components of health and disease. Disruption of the oral microbiome has been proven to influence the course of oral diseases, especially among immunocompromised patients. Oral microbiome is comprised of inter-kingdom microorganisms, including yeasts such as Candida albicans, bacteria, archaea and viruses. These microorganisms can interact synergistically, mutualistically and antagonistically, wherein the sum of these interactions dictates the composition of the oral microbiome. For instance, polymicrobial interactions can improve the ability of C albicans to form biofilm, which subsequently increases the colonisation of oral mucosa by the yeast. Polymicrobial interactions of C albicans with other members of the oral microbiome have been reported to enhance the malignant phenotype of oral cancer cells, such as the attachment to extracellular matrix molecules (ECM) and epithelial-mesenchymal transition (EMT). Polymicrobial interactions may also exacerbate an inflammatory response in oral epithelial cells, which may play a role in carcinogenesis. This review focuses on the role of polymicrobial interactions between C albicans and other oral microorganisms, including its role in promoting oral carcinogenesis.
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    Feasibility and development of a cariogenic diet scale for epidemiological research
    Amezdroz, E ; Carpenter, L ; Johnson, S ; Flood, V ; Dashper, SG ; Calache, H ; Gussy, M ; Waters, E (WILEY, 2019-05)
    BACKGROUND: Diet cariogenicity plays a major role as both a protective and risk factor in the development of early childhood caries (ECC). AIM: Develop a scale measuring the cariogenicity of foods and beverages and employ it to describe the cariogenicity of young children's diets and predict dental caries outcomes. DESIGN: Scores of cariogenicity and consumption frequency were applied to food frequency questionnaire (FFQ) collected from an Australian children's cohort study with three time-points of data. One-way ANOVA, with post hoc Tukey test compared mean cariogenic scale measured at 18 months between the subsample of children with caries classification at age 5 years. RESULTS: At 6 months, children's mean cariogenic score was 10.05, increasing to 34.18 at 12 and 50.00 at 18 months. Mean cariogenic scale score at 18 months was significantly higher in children with advanced disease at 5 years (mean scale score: 59.0 ± 15.9) compared to those that were healthy (mean score 47.7 ± 17.5, P = 0.007) or had mild-moderate disease (mean score 48.2 ± 17.3, P = 0.008). CONCLUSIONS: The cariogenic diet scale provides a useful indication of the increasing cariogenicity of children's diets with age and highlights the incorporation of discretionary choice foods and beverages into the diets of young children much earlier than nutritionally recommended.
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    Incorporation of the microencapsulated antimicrobial agent phytoncide into denture base resin
    An, S ; Judge, RB ; Wong, RH ; Arzmi, MH ; Palamara, JE ; Dashper, SG (WILEY, 2018-09)
    BACKGROUND: This study aimed to fabricate a denture base resin (DBR) containing phytoncide microcapsules (PTMCs) and determine the mechanical properties of the resin and antifungal activity. METHODS: Fifty-four heat-cured rectangular DBR specimens (64 × 10 × 3.3 ± 0.2 mm) containing nine concentrations of PTMC between 0 and 5% (wt/wt) were fabricated and subjected to a three-point bending test. A phytoncide release bioassay was developed using DBR containing 0% and 2.5% PTMCs (wt/wt) in a 24 well-plate assay with incubation of Porphyromonas gingivalis at 37 °C for 74 h. The antifungal activity of PTMCs against Candida albicans, in a pH 5.5 acidic environment was determined in a plate assay. RESULTS: Flexural strength decreased with increasing PTMC concentration from 97.58 ± 4.79 MPa for the DBR alone to 53.66 ± 2.46 MPa for DBR containing 5.0% PTMC. No release of phytoncide from the PTMCs in the DBR was detected at pH 7.4. The PTMCs had a minimal inhibitory concentration of 2.6% (wt/vol) against C. albicans at pH 5.5. CONCLUSIONS: PTMCs can be added to DBR 2.5% (wt/wt) without adversely affecting flexural strength. PTMCs released the antimicrobial agent at pH 5.5 at concentrations sufficient to inhibit the growth of the C. albicans.
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    The VicGeneration study - a birth cohort to examine the environmental, behavioural and biological predictors of early childhood caries: background, aims and methods
    de Silva-Sanigorski, AM ; Calache, H ; Gussy, M ; Dashper, S ; Gibson, J ; Waters, E (BMC, 2010-02-25)
    BACKGROUND: Dental caries (decay) during childhood is largely preventable however it remains a significant and costly public health concern, identified as the most prevalent chronic disease of childhood. Caries in children aged less than five years (early childhood caries) is a rapid and progressive disease that can be painful and debilitating, and significantly increases the likelihood of poor child growth, development and social outcomes. Early childhood caries may also result in a substantial social burden on families and significant costs to the public health system. A disproportionate burden of disease is also experienced by disadvantaged populations. METHODS/DESIGN: This study involves the establishment of a birth cohort in disadvantaged communities in Victoria, Australia. Children will be followed for at least 18 months and the data gathered will explore longitudinal relationships and generate new evidence on the natural history of early childhood caries, the prevalence of the disease and relative contributions of risk and protective biological, environmental and behavioural factors. Specifically, the study aims to:1. Describe the natural history of early childhood caries (at ages 1, 6, 12 and 18 months), tracking pathways from early bacterial colonisation, through non-cavitated enamel white spot lesions to cavitated lesions extending into dentine.2. Enumerate oral bacterial species in the saliva of infants and their primary care giver.3. Identify the strength of concurrent associations between early childhood caries and putative risk and protective factors, including biological (eg microbiota, saliva), environmental (fluoride exposure) and socio-behavioural factors (proximal factors such as: feeding practices and oral hygiene; and distal factors such as parental health behaviours, physical health, coping and broader socio-economic conditions).4. Quantify the longitudinal relationships between these factors and the development and progression of early childhood caries from age 1-18 months. DISCUSSION: There is currently a lack of research describing the natural history of early childhood caries in very young children, or exploring the interactions between risk and protective factors that extend to include contemporary measures of socio-behavioural factors. This study will generate knowledge about pathways, prevalence and preventive opportunities for early childhood caries, the most prevalent child health inequality.
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    Mechanistic Target of Rapamycin (Mtor) Is Essential for Murine Embryonic Heart Development and Growth
    Zhu, Y ; Pires, KMP ; Whitehead, KJ ; Olsen, CD ; Wayment, B ; Zhang, YC ; Bugger, H ; Ilkun, O ; Litwin, SE ; Thomas, G ; Kozma, SC ; Abel, ED ; Brissette, CA (PUBLIC LIBRARY SCIENCE, 2013-01-14)
    Chronic periodontitis has a polymicrobial biofilm aetiology and interactions between key bacterial species are strongly implicated as contributing to disease progression. Porphyromonas gingivalis, Treponema denticola and Tannerella forsythia have all been implicated as playing roles in disease progression. P. gingivalis cell-surface-located protease/adhesins, the gingipains, have been suggested to be involved in its interactions with several other bacterial species. The aims of this study were to determine polymicrobial biofilm formation by P. gingivalis, T. denticola and T. forsythia, as well as the role of P. gingivalis gingipains in biofilm formation by using a gingipain null triple mutant. To determine homotypic and polymicrobial biofilm formation a flow cell system was employed and the biofilms imaged and quantified by fluorescent in situ hybridization using DNA species-specific probes and confocal scanning laser microscopy imaging. Of the three species, only P. gingivalis and T. denticola formed mature, homotypic biofilms, and a strong synergy was observed between P. gingivalis and T. denticola in polymicrobial biofilm formation. This synergy was demonstrated by significant increases in biovolume, average biofilm thickness and maximum biofilm thickness of both species. In addition there was a morphological change of T. denticola in polymicrobial biofilms when compared with homotypic biofilms, suggesting reduced motility in homotypic biofilms. P. gingivalis gingipains were shown to play an essential role in synergistic polymicrobial biofilm formation with T. denticola.
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    Porphyromonas gingivalis Peptidylarginine Deiminase, a Key Contributor in the Pathogenesis of Experimental Periodontal Disease and Experimental Arthritis
    Gully, N ; Bright, R ; Marino, V ; Marchant, C ; Cantley, M ; Haynes, D ; Butler, C ; Dashper, S ; Reynolds, E ; Bartold, M ; Yilmaz, Ö (PUBLIC LIBRARY SCIENCE, 2014-06-24)
    OBJECTIVES: To investigate the suggested role of Porphyromonas gingivalis peptidylarginine deiminase (PAD) in the relationship between the aetiology of periodontal disease and experimentally induced arthritis and the possible association between these two conditions. METHODS: A genetically modified PAD-deficient strain of P. gingivalis W50 was produced. The effect of this strain, compared to the wild type, in an established murine model for experimental periodontitis and experimental arthritis was assessed. Experimental periodontitis was induced following oral inoculation with the PAD-deficient and wild type strains of P. gingivalis. Experimental arthritis was induced via the collagen antibody induction process and was monitored by assessment of paw swelling and micro-CT analysis of the radio-carpal joints. Experimental periodontitis was monitored by micro CT scans of the mandible and histological assessment of the periodontal tissues around the mandibular molars. Serum levels of anti-citrullinated protein antibodies (ACPA) and P. gingivalis were assessed by ELISA. RESULTS: The development of experimental periodontitis was significantly reduced in the presence of the PAD-deficient P. gingivalis strain. When experimental arthritis was induced in the presence of the PAD-deficient strain there was less paw swelling, less erosive bone damage to the joints and reduced serum ACPA levels when compared to the wild type P. gingivalis inoculated group. CONCLUSION: This study has demonstrated that a PAD-deficient strain of P. gingivalis was associated with significantly reduced periodontal inflammation. In addition the extent of experimental arthritis was significantly reduced in animals exposed to prior induction of periodontal disease through oral inoculation of the PAD-deficient strain versus the wild type. This adds further evidence to the potential role for P. gingivalis and its PAD in the pathogenesis of periodontitis and exacerbation of arthritis. Further studies are now needed to elucidate the mechanisms which drive these processes.
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    Natural history of dental caries in very young Australian children
    Gussy, M ; Ashbolt, R ; Carpenter, L ; Virgo-Milton, M ; Calache, H ; Dashper, S ; Leong, P ; de Silva, A ; de Livera, A ; Simpson, J ; Waters, E (WILEY, 2016-05)
    BACKGROUND: Whilst the global burden of caries is increasing, the trajectory of decay in young children and the point at which prevention should occur has not been well established. AIM: To identify the 'natural history' of dental caries in early childhood. DESIGN: A birth cohort study was established with 467 mother/child dyads followed at 1, 6, 12, 18, and 36 months of age. Parent-completed surveys captured demographic, social, and behavioural data, and oral examinations provided clinical and data. RESULTS: Eight per cent of children (95% confidence interval (CI): 5-12%) at 18 months and 23% (95% CI: 18-28%) at 36 months experienced decay. Interesting lesion behaviour was found between 18 and 36 months, with rapid development of new lesions on sound teeth (70% of teeth, 95% CI: 63-76%) and regression of many lesions from non-cavitated lesions to sound (23% of teeth, 95% CI: 17-30%). Significant associations were found between soft drink consumption and lesion progression. CONCLUSIONS: Findings suggest optimal time periods for screening and prevention of a disease which significantly impacts multiple health and well-being outcomes across the life course.
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    Temporal development of the oral microbiome and prediction of early childhood caries
    Dashper, SG ; Mitchell, HL ; Le Cao, K-A ; Carpenter, L ; Gussy, MG ; Calache, H ; Gladman, SL ; Bulach, DM ; Hoffmann, B ; Catmull, D ; Pruilh, S ; Johnson, S ; Gibbs, L ; Amezdroz, E ; Bhatnagar, U ; Seemann, T ; Mnatzaganian, G ; Manton, DJ ; Reynolds, EC (NATURE PUBLISHING GROUP, 2019-12-24)
    Human microbiomes are predicted to assemble in a reproducible and ordered manner yet there is limited knowledge on the development of the complex bacterial communities that constitute the oral microbiome. The oral microbiome plays major roles in many oral diseases including early childhood caries (ECC), which afflicts up to 70% of children in some countries. Saliva contains oral bacteria that are indicative of the whole oral microbiome and may have the ability to reflect the dysbiosis in supragingival plaque communities that initiates the clinical manifestations of ECC. The aim of this study was to determine the assembly of the oral microbiome during the first four years of life and compare it with the clinical development of ECC. The oral microbiomes of 134 children enrolled in a birth cohort study were determined at six ages between two months and four years-of-age and their mother's oral microbiome was determined at a single time point. We identified and quantified 356 operational taxonomic units (OTUs) of bacteria in saliva by sequencing the V4 region of the bacterial 16S RNA genes. Bacterial alpha diversity increased from a mean of 31 OTUs in the saliva of infants at 1.9 months-of-age to 84 OTUs at 39 months-of-age. The oral microbiome showed a distinct shift in composition as the children matured. The microbiome data were compared with the clinical development of ECC in the cohort at 39, 48, and 60 months-of-age as determined by ICDAS-II assessment. Streptococcus mutans was the most discriminatory oral bacterial species between health and current disease, with an increased abundance in disease. Overall our study demonstrates an ordered temporal development of the oral microbiome, describes a limited core oral microbiome and indicates that saliva testing of infants may help predict ECC risk.