Comparative effects of various commercially available mouthrinse formulations on oral malodor.

OBJECTIVES: The primary aim of this study was to compare a new mouthwash (SB12®) containing 0.025% chlorhexidine and 0.3% zinc for oral malodor reduction against four commercially available mouthwashes and negative control. A secondary aim was to compare the two methods for measuring volatile sulphur compounds (VSC) by halimetry and OralChroma. METHODS: Organoleptic scale, halimeter and the OralChroma were used to assess oral malodour and VSC. The effects of five test formulations and water (negative control) were assessed after 30, 60, 90 and 180 min, with 1 week between the treatments to avoid any cross-over effect. RESULTS: Reduction in H(2) S by halimetry and malodour levels by organoleptic assessment ranged from, slight (LacerFresh®) (P > 0.05), moderate (BreathRx®, SmartMouth® (P < 0.01) to marked effects (SB12®, Listerine®) (P < 0.001) at all time points compared with water. The largest differences were observed at 30 min and decreased with time. SB12® showed separation from Listerine® at 180 min, using ANOVA plus Bonferroni's Multiple Comparison post-test (P < 0.05). Relationships between organoleptic, halimeter and OralChroma were between R² = 0.795 and 0.926. CONCLUSION: SB12 shows a consistent and reproducible inhibitory effect on oral malodor parameters, which in turn correlate well with each other.

In vitro comparison of antimicrobial activity of iodine and silver dressings against biofilms.

OBJECTIVE: To compare the antimicrobial effectiveness of silver- and iodine-containing wound dressings against preformed mature biofilms of pathogenic wound bacteria grown in vitro. METHOD: Biofilms of Pseudomonas aeruginosa and Staphylococcus aureus were grown within an in vitro flat bed perfusion biofilm model. Mature biofilms were removed and exposed to wound dressings containing either silver or iodine (Aquacel Ag and Iodozyme) within a static diffusion method, for up to 24 hours. This method was designed to reflect certain key features that determine antimicrobial activity within the wound. The numbers of viable bacteria surviving in the biofilms were determined at set time intervals over the test period. RESULTS: Both test dressings exerted an antimicrobial effect against the target species biofilms, although the iodine dressing was more efficacious under the experimental conditions employed. CONCLUSION: There are large and potentially significant differences (as measured in vitro) in the effectiveness of wound dressings containing broad-spectrum antimicrobial agents such as silver and iodine against specific types of bacterial biofilms.

Use of a bioluminescent Pseudomonas aeruginosa strain within an in vitro microbiological system, as a model of wound infection, to assess the antimicrobial efficacy of wound dressings by monitoring light production.

A bioluminescent Pseudomonas aeruginosa was incorporated into an in vitro static diffusion method to determine whether light output could be used as a measure of wound dressing efficacy. A significant linear correlation was observed between viable counts and bioluminescence during exponential growth in planktonic culture (r(2) = 0.969). Exponential-phase cells were used to inoculate cellulose discs for integration into an in vitro wound model that incorporated a reservoir of serum. A significant linear correlation was found between bioluminescence (photon counts monitored by a low-light camera) and viable counts in this growth environment (r(2) = 0.982). Three antimicrobial wound dressings were applied to the surface of freshly prepared sample discs within the wound model, and the kill kinetics were codetermined by photon and viable counts. Quantifiable kill rates gave the same order of efficacy for the three wound dressings using both types of measurement, and a significant linear correlation was shown between photon and viable counts (r(2) = 0.873) within this killing environment. Under all defined conditions, a significant linear correlation between bioluminescence and viable counts was shown but the actual slope of the correlation was different, depending on the physicochemical environment of the cells. Hence, significantly more light per cell (P < 0.0001) was produced when cells in discs were exposed to a killing environment compared to a growing environment. As long as defined conditions are employed, the resulting linear correlation enables the state of the system to be continually monitored without disturbance, allowing more immediate and accurate calculations of kill rates without the need for viable counting.

An in vitro study of antimicrobial activity and efficacy of iodine-generating hydrogel dressings.

OBJECTIVE: To determine the antimicrobial activity and efficacy of different formulations of novel bioxygenating hydrogel dressings (which deliver both iodine and oxygen into the wound) against various target organisms by means of an in vitro test system that more effectively mimics the conditions encountered when dressings are in contact with wounds. METHOD: Three bioxygenating hydrogels were tested: Oxyzyme, which releases low levels of iodine into the wound, and Iodozyme 402 and Iodozyme 401, which release higher levels of iodine, with Iodozyme 402 releasing twice the amount of 401. Cellulose filter disks (n = 32) were inoculated with indicator species and placed equidistant from each other as a matrix onto agar test beds. Cut squares of control or test dressings were placed on top of each disk. Kill curves were constructed from determinations of the numbers of survivors (log cfu per disk) over time by removing disk samples at various time points. RESULTS: Significant differences (p < 0.05) were observed between the controls and test samples. The order of sensitivity for Oxyzyme was Fusobacterium nucleatum, Bacteroides fragilis, Propionibacterium acnes, Staphylococcus epidermidis, methicillin-resistant Staphylococcus aureus (MRSA), Candida albicans and Pseudomonas aeruginosa. The order of efficacy of the three hydrogel dressings (Iodozyme 402, followed by Iodozyme 401 and then Oxyzyme) was the same regardless of the target species. CONCLUSION: The novel hydrogel skin surface wound dressings are broad-spectrum in activity, encompassing antibiotic-resistant organisms, anaerobes and yeasts; their antimicrobial function appears to be rapidly effective.

Antimicrobial photodynamic therapy: assessment of genotoxic effects on keratinocytes in vitro.

BACKGROUND: Work has shown that cutaneous microbial species associated with skin conditions of microbial aetiology are susceptible to killing by antimicrobial photodynamic therapy (APDT) using visible light and methylene blue. OBJECTIVES: To evaluate immediate and delayed genotoxicity of APDT on keratinocytes in vitro. METHODS: A combination of methylene blue (100 microg mL(-1)) and visible light (42 mW cm(-2)), as used in studies of microbe and keratinocyte cytotoxicity, was employed to test a human keratinocyte cell line (H103) for genotoxic damage by comet assay. RESULTS: The comet assay was able to detect genotoxic damage in H2O2-treated keratinocytes (positive control). APDT did not cause either immediate or delayed genotoxic damage in keratinocytes following APDT of up to 180 min. CONCLUSIONS: APDT sufficient to reduce microbes by seven log cycles showed no detectable genotoxic effects on keratinocytes. APDT applied in vivo may represent a useful low-risk alternative to conventional antimicrobial treatment in dermatology.

Biofilm formation by Helicobacter pylori.

Helicobacter pylori NCTC 11637 produces a water-insoluble biofilm when grown under defined conditions with a high carbon:nitrogen ratio in continuous culture and in 10% strength Brucella broth supplemented with 3 g l-1 glucose. Biofilm accumulated at the air/liquid interface of the culture. Light microscopy of frozen sections of the biofilm material showed few bacterial cells in the mass of the biofilm. The material stained with periodic acid Schiff's reagent. Fucose, glucose, galactose, and glycero-manno-heptose, N-acetylglucosamine and N-acetylmuramic acid were identified in partially purified and in crude material, using gas chromatography and mass spectrometry. The sugar composition strongly indicates the presence of a polysaccharide as a component of the biofilm material. Antibodies (IgG) to partially purified material were found in both sero-positive and sero-negative individuals. Treatment of the biofilm material with periodic acid reduced or abolished immunoreactivity. Treatment with 5 mol l-1 urea at 100 degrees C and with phenol did not remove antigenic recognition by patient sera. The production of a water-insoluble biofilm by H. pylori may be important in enhancing resistance to host defence factors and antibiotics, and in microenvironmental pH homeostasis facilitating the growth and survival of H. pylori in vivo.