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result(s) for
"Pratten, Jonathan"
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An In Vitro Evaluation of Denture Cleansing Regimens against a Polymicrobial Denture Biofilm Model
by
Young, Tracy
,
Ramage, Gordon
,
Pratten, Jonathan R.
in
antimicrobials
,
Bacteria
,
biofilm model
2022
Denture stomatitis (DS) is an inflammatory disease resulting from a polymicrobial biofilm perturbation at the denture surface–palatal mucosa interface. Recommendations made by dental health care professionals often lack clarity for appropriate denture cleaning. This study investigated the efficacy of brushing with off-the-shelf denture cleanser (DC) tablets (Poligrip®) vs. two toothpastes (Colgate® and Crest®) in alleviating the viable microorganisms (bacteria and fungi) in an in vitro denture biofilm model. Biofilms were grown on poly(methyl)methacrylate (PMMA) discs, then treated daily for 7 days with mechanical disruption (brushing), plus Poligrip® DC, Colgate® or Crest® toothpastes. Weekly treatment with Poligrip® DC on day 7 only was compared to daily modalities. All treatment parameters were processed to determine viable colony forming units for bacteria and fungi using the Miles and Misra technique, and imaged by confocal laser scanning microscopy (CLSM). Brushing with daily DC therapy was the most effective treatment in reducing the viable biofilm over 7 days of treatment. Brushing only was ineffective in controlling the viable bioburden, which was confirmed by CLSM imaging. This data indicates that regular cleansing of PMMA with DC was best for polymicrobial biofilms.
Journal Article
Biological Evaluation of Oral Care Products Using 3D Tissue-Engineered In Vitro Models of Plaque-Induced Gingivitis
by
Franklin, Kirsty
,
Pratten, Jonathan
,
Moharamzadeh, Keyvan
in
Antibacterial agents
,
Antibiotics
,
Antiinfectives and antibacterials
2024
Background: The aim of this study was to investigate and visualize the anti-inflammatory and anti-bacterial effects of different oral care products using an infected and inflamed 3D tissue-engineered gingival mucosal model. Methods: A 3D full-thickness oral mucosal model was engineered inside tissue culture inserts using collagen hydrogels populated with human gingival fibroblasts and THP-1 monocytes and layered with oral epithelial cell lines. Oral saliva bacteria were cultured and added to the surface of the models and inflammation was further simulated with lipopolysaccharide (LPS) of Escherichia coli. The 3D models were exposed to three different types of toothpastes, a chlorhexidine antiseptic mouthwash, different antibiotics, and a mechanical rinse with phosphate-buffered saline (PBS) prior to biological evaluation using the PrestoBlue tissue viability assay, histology, optical coherence tomography (OCT), confocal microscopy, and measurement of the release of the inflammatory markers IL-1β, IL-6, and IL-8 with ELISA. Results: Multiple-endpoint analyses of the infected oral mucosal models treated with different anti-bacterial agents showed consistent outcomes in terms of tissue viability, histology, OCT, and confocal microscopy findings. In terms of anti-inflammatory testings, the positive control group showed the highest level of inflammation compared with all other groups. Depending on the anti-bacterial and anti-inflammatory potential of the test groups, different levels of inflammation were observed in the test groups. Conclusions: The inflamed 3D oral mucosal model developed in this study has the potential to be used as a suitable in vitro model for testing the biocompatibility, anti-inflammatory, and anti-bacterial properties of oral care products including mouthwashes and toothpastes. The results of this study indicate that the chlorhexidine mouthwash has both anti-bacterial and cytotoxic effects on the 3D oral mucosal model. Hyaluronic-acid-containing toothpaste has significant anti-bacterial and anti-inflammatory effects on the 3D oral mucosal model.
Journal Article
Candida albicans as an Essential “Keystone” Component within Polymicrobial Oral Biofilm Models?
by
Young, Tracy
,
Alshanta, Om-Alkhir
,
Ramage, Gordon
in
anti-infective agents
,
Antimicrobial agents
,
antimicrobials
2020
Background: Existing standardized biofilm assays focus on simple mono-species or bacterial-only models. Incorporating Candida albicans into complex biofilm models can offer a more appropriate and relevant polymicrobial biofilm for the development of oral health products. Aims: This study aimed to assess the importance of interkingdom interactions in polymicrobial oral biofilm systems with or without C. albicans, and test how these models respond to oral therapeutic challenges in vitro. Materials and Methods: Polymicrobial biofilms (two models containing 5 and 10 bacterial species, respectively) were created in parallel in the presence and absence of C. albicans and challenged using clinically relevant antimicrobials. The metabolic profiles and biomasses of these complex biofilms were estimated using resazurin dye and crystal violet stain, respectively. Quantitative PCR was utilized to assess compositional changes in microbial load. Additional assays, for measurements of pH and lactate, were included to monitor fluctuations in virulence “biomarkers.” Results: An increased level of metabolic activity and biomass in the presence of C. albicans was observed. Bacterial load was increased by more than a factor of 10 in the presence of C. albicans. Assays showed inclusion of C. albicans impacted the biofilm virulence profiles. C. albicans did not affect the biofilms’ responses to the short-term incubations with different treatments. Conclusions: The interkingdom biofilms described herein are structurally robust and exhibit all the hallmarks of a reproducible model. To our knowledge, these data are the first to test the hypothesis that yeasts may act as potential “keystone” components of oral biofilms.
Journal Article
Susceptibility of MRSA biofilms to denture-cleansing agents
by
Pratten, Jonathan
,
Wilson, Michael
,
Ready, Derren
in
Acrylic resins
,
antibiotic resistance
,
Antibiotics, microbial producers, chemotherapic agents, antiseptics, disinfecting agents
2009
Methicillin-resistant Staphylococcus aureus (MRSA) is an important nosocomial pathogen, which is responsible for considerable morbidity and mortality in the United Kingdom. The major reservoir of this organism is thought to be the anterior nares, but there is increasing evidence that this pathogen is present in the oral cavity, particularly in denture wearers. The purpose of this study was to determine whether MRSA, grown as biofilms on denture acrylic resin, could be eradicated using commercially available agents. EMRSA-15 or EMRSA-16 was grown in a model system on the surface of denture acrylic resin for 4, 24 or 120 h before the samples were exposed to a range of disinfectants for time intervals of 1, 5 and 10 min. All of the agents reduced the number of cultivable MRSA bacteria present on the acrylic resin surface at 4 h, with 2% sodium hypochlorite (NaOCl) eliminating MRSA below the level of detection after an exposure of 1 min. However, the established MRSA biofilms (24 and 120 h) were more resistant to killing by the agents, although 2% NaOCl was still able to eradicate all ages of MRSA biofilms within 1 min of exposure.
Journal Article
Frictional properties of light-activated antimicrobial polymers in blood vessels
by
Prokopovich, Polina
,
Perni, Stefano
,
Pratten, Jonathan
in
Animals
,
Anti-Infective Agents - chemistry
,
Anti-Infective Agents - radiation effects
2010
The adhesion of microbes to catheter surfaces is a serious problem and the resulting infections frequently lead to longer hospitalisation and higher risk for the patient. Several approaches have been developed to produce materials that are less susceptible to microbial colonisation. One such approach is the incorporation of photoactivated compounds, such as Toluidine Blue O (TBO), in the polymeric matrix resulting in ‘light-activated antimicrobial materials’. The insertion and removal of catheters can cause tissue damage and patient discomfort through frictional forces; hence the lubricity of a catheter material is also very important. In this work the tribological performance of silicone and polyurethane containing TBO and gold nanoparticles were evaluated using two different surfaces, the inner part of the aorta and the superior vena cava of sheep. Static and kinetic friction coefficients of these materials were measured using a tribometric device developed for in vitro applications using dry materials and those lubricated with blood. It was found that neither the preparation process nor the presence of TBO or gold nanoparticles, had an effect on the friction factors in comparison to those of untreated materials. In all cases, static and kinetic friction coefficients on aorta tissue were higher than those on vena cava due to higher surface roughness of the aorta. The presence of blood as a lubricant resulted in lower friction coefficients.
Journal Article
Composition of in vitro denture plaque biofilms and susceptibility to antifungals
by
Lamfon, Hanadi
,
Al-Karaawi, Zubaida
,
McCullough, Michael
in
Aged
,
Antifungal agents
,
Antifungal Agents - pharmacology
2005
The aim of this study was to investigate the composition of microcosm denture plaque biofilms and the susceptibility of
Candida spp. within these biofilms to antifungal agents. An in vitro model was employed to grow oral biofilms derived from denture associated stomatitis (DAS) patient samples to assess fungal growth in the presence and absence of antifungal agents. The compositions of genera present in vitro were found to be similar to those exhibited on the mucosa and denture fitting surfaces of DAS samples. Exposure to single agents, e.g., miconazole, fluconazole or chlorhexidine did not inhibit growth of
Candida spp. when used in clinically relevant doses. Combinations of miconazole and chlorhexidine, pulsed into the system to mimic patient use, did reduce bacterial and candidal growth for several days. Hence, the use of dual-therapy appeared to be useful in reducing the number of viable organisms within denture plaque grown in vitro although resistance to these agents was also evident.
Journal Article
Effects of Mushroom and Chicory Extracts on the Physiology and Shape of Prevotella intermedia, a Periodontopathogenic Bacterium
by
Signoretto, Caterina
,
Spratt, David A.
,
Bertoncelli, Anna
in
Agaricales
,
Agaricales - chemistry
,
Anti-Bacterial Agents
2011
Contrary to the common assumption that food has a negative impact on oral health, research has shown that several foods contain a number of components with antibacterial and antiplaque activity. These natural compounds may be useful for improving daily oral hygiene. In this study we evaluate the mode of antimicrobial action of fractions of mushroom and red chicory extracts on Prevotella intermedia, a periodontopathogenic bacterium. The minimal inhibitory concentration corresponded to 0.5x compared to the natural food concentration for both extracts. This concentration resulted in a bacteriostatic effect in mushroom extract and in a slightly bactericidal effect in chicory extract. Cell mass continued to increase even after division stopped. As regards macromolecular synthesis, DNA was almost totally inhibited upon addition of either mushroom or chicory extract, and RNA to a lesser extent, while protein synthesis continued. Cell elongation occurred after septum inhibition as documented by scanning electron microscopy and cell measurement. The morphogenetic effects are reminiscent of the mode of action of antibiotics such as quinolones or β-lactams. The discovery of an antibiotic-like mode of action suggests that these extracts can be advantageously employed for daily oral hygiene in formulations of cosmetic products such as mouthwashes and toothpastes.
Journal Article
Development of an in vitro periodontal biofilm model for assessing antimicrobial and host modulatory effects of bioactive molecules
by
Ramage, Gordon
,
Middleton, Andrew M
,
Pratten, Jonathan
in
Aggregatibacter actinomycetemcomitans - drug effects
,
Anti-Bacterial Agents - pharmacology
,
Anti-Bacterial Agents - toxicity
2014
Inflammation within the oral cavity occurs due to dysregulation between microbial biofilms and the host response. Understanding how different oral hygiene products influence inflammatory properties is important for the development of new products. Therefore, creation of a robust host-pathogen biofilm platform capable of evaluating novel oral healthcare compounds is an attractive option. We therefore devised a multi-species biofilm co-culture model to evaluate the naturally derived polyphenol resveratrol (RSV) and gold standard chlorhexidine (CHX) with respect to anti-biofilm and anti-inflammatory properties.
An in vitro multi-species biofilm containing S. mitis, F. nucleatum, P. gingivalis and A. actinomycetemcomitans was created to represent a disease-associated biofilm and the oral epithelial cell in OKF6-TERT2. Cytotoxicity studies were performed using RSV and CHX. Multi-species biofilms were either treated with either molecule, or alternatively epithelial cells were treated with these prior to biofilm co-culture. Biofilm composition was evaluated and inflammatory responses quantified at a transcriptional and protein level.
CHX was toxic to epithelial cells and multi-species biofilms at concentrations ranging from 0.01-0.2%. RSV did not effect multi-species biofilm composition, but was toxic to epithelial cells at concentrations greater than 0.01%. In co-culture, CHX-treated biofilms resulted in down regulation of the inflammatory chemokine IL-8 at both mRNA and protein level. RSV-treated epithelial cells in co-culture were down-regulated in the release of IL-8 protein, but not mRNA.
CHX possesses potent bactericidal properties, which may impact downstream inflammatory mediators. RSV does not appear to have bactericidal properties against multi-species biofilms, however it did appear to supress epithelial cells from releasing inflammatory mediators. This study demonstrates the potential to understand the mechanisms by which different oral hygiene products may influence gingival inflammation, thereby validating the use of a biofilm co-culture model.
Journal Article
Novel Light-Activated Antimicrobial Coatings Are Effective Against Surface-Deposited Staphylococcus aureus
by
Decraene, Valérie
,
Pratten, Jonathan
,
Wilson, Michael
in
Aerosols
,
analogs & derivatives
,
Animals
2008
Aerosols constitute a major route of transmission for a wide range of infectious diseases in the hospital setting. The aim of this study was to determine the survival of Staphylococcus aureus on a light-activated antimicrobial coating. S. aureus suspended in phosphate-buffered saline (PBS), saliva, or horse serum was sprayed onto cellulose acetate coatings containing toluidine blue O and rose bengal and the survival of the organism on these surfaces was determined following 6 h of exposure to a 28-W domestic fluorescent lamp (light intensity = 3700 ± 20 lux). Kills ranging from 78.9% (in horse serum) to 99.8% (in PBS) were obtained when the bacterial density on the coatings was ~10⁵ colony-forming units/m². The results of this study have shown that a coating containing toluidine blue and rose bengal can achieve significant kills of S. aureus when illuminated by a domestic light source. Light-activated coatings could provide a simple, low-cost means of reducing the microbial load in hospitals and other facilities.
Journal Article
Antibacterial Activity of Light-Activated Silicone Containing Methylene Blue and Gold Nanoparticles of Different Sizes
by
Wilson, Mike
,
Uppal, Madeeha
,
Pratten, Jonathan
in
Antimicrobial agents
,
Aqueous solutions
,
Bacteria
2010
Methylene Blue and methylene blue-gold nanoparticle mixtures were encapsulated in a silicone polymer using a swell-encapsulation-shrink technique. The antibacterial properties of the materials, when tested against
Escherichia coli
and
Staphylococcus epidermidis
, and exposed to laser light (660 nm), were significantly affected by both the presence and size of Au nanoparticles. Bacterial inactivation data were analysed using the Weibull inactivation model. For both
E. coli
and
S. epidermidis
the value of the parameter, indicating the time required to achieve the first log
10
reduction in the viable count, decreased when Au nanoparticles of ca 2 nm diameter were present. Larger Au nanoparticles (diameters of 5 and 20 nm) in combination with methylene blue were also embedded in silicone. The values of these materials increased with nanoparticle diameter, indicating a reduction in antibacterial activity. In all cases
E. coli
had higher values than
S. epidermidis
.
Journal Article