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The penetration of biofilms by antimicrobials is a potential limiting factor in biofilm control. This is relevant to oral health, where compounds used to control microbial growth and activities could also affect the permeability of dental plaque biofilm with secondary effects on biofilm tolerance. We have investigated the effects of zinc salts on the permeability of Streptococcus mutans biofilms. Biofilms were grown with low concentrations of zinc acetate-(ZA) and a transwell transportation assay was applied to test biofilm permeability in an apical-basolateral direction. Crystal violet assays and total viable counts were used to quantify biofilm formation and viability, respectively, and short timeframe diffusion rates within microcolonies were determined using spatial Intensity Distribution Analysis (SpIDA). Whilst diffusion rates within biofilm microcolonies were not significantly altered, exposure to ZA significantly increased the overall permeability of S. mutans biofilms (p

Aims Caries and periodontal disease are associated with inadequate control of oral bacteria. Since conventional microbiological evaluations are impractical in dental clinics or public engagement activities, a rapid test for the quantification of oral bacteria represents a useful tool. We describe the development of a colour change test to rapidly estimate bacterial colonisation density in the mouth. Methods and results Volunteers rinsed with milk or milkshake. Viability indicators were added and colour changes quantified during incubation. Using milkshake and the resazurin-based solution PrestoBlue (9% v/v), the method distinguished between samples before and after brushing within 5 min. Colour changes were quantified and viable counts were obtained using oral rinses. Measured colour changes strongly correlated with total counts of both anaerobes and streptococci (Spearman's correlation coefficient of 0·782 and 0·769, respectively, P ≤ 0·001) and with perceived changes, as determined by volunteers (n = 10) visually ranking images. Conclusions The resazurin milkshake test can rapidly and visually quantify viable bacteria in oral samples. Significance and impact of the study The resazurin milkshake test could serve as a sensitive semi-quantitative method for measuring oral bacteria in human oral rinse samples.

Small proteinaceous oligomeric species contribute to the formation of larger aggregates, a phenomenon that is of direct relevance to the characterization of protein aggregation in biopharmaceuticals and understanding the underlying processes contributing to neurodegenerative diseases.The ability to monitor in situ oligomerization and aggregation processes renders imaging and image analysis an attractive approach for gaining a mechanistic insight into early processes contributing to the formation of larger aggregates in disease models and biologics. The combination of image analysis tools enables the detection of both oligomeric and larger aggregate subtype in contrast to conventional kinetic-based approaches that lack the ability to resolve dimers from monomeric moieties in samples containing mixed populations.In this chapter, we describe the process for confocal time series image acquisition for monitoring the in situ loss of monomers, and the subsequent analysis pipeline using spatial intensity distribution analysis (SpIDA) to evaluate oligomer content.

Small proteinaceous oligomeric species contribute to the formation of larger aggregates, a phenomenon that is of direct relevance to the characterization of protein aggregation in biopharmaceuticals and understanding the underlying processes contributing to neurodegenerative diseases. The ability to monitor in situ oligomerization and aggregation processes renders imaging and image analysis an attractive approach for gaining a mechanistic insight into early processes contributing to the formation of larger aggregates in disease models and biologics. The combination of image analysis tools enables the detection of both oligomeric and larger aggregate subtype in contrast to conventional kinetic-based approaches that lack the ability to resolve dimers from monomeric moieties in samples containing mixed populations. In this chapter, we describe the process for confocal time series image acquisition for monitoring the in situ loss of monomers, and the subsequent analysis pipeline using spatial intensity distribution analysis (SpIDA) to evaluate oligomer content.

Background\ud \ud Cetylpyridinium chloride (CPC) and sodium fluoride augment oral hygiene by inactivating bacteria and inhibiting enamel demineralisation, respectively. However, there are few reports in the literature documenting the antibacterial efficacy of their combined use in mouthrinses. We have used six experimental systems to compare the antibacterial effects of mouthrinses containing 0.075 % CPC (test rinse, TR) or 0.075 % CPC with sodium fluoride (test fluoride rinse, TFR).\ud \ud \ud Results\ud \ud Effects against planktonic bacteria were determined using viable counting (for Streptococcus mutans and salivary bacteria), a redox dye (for Actinomyces viscosus and salivary bacteria) and viable counting (for ex vivo oral rinses). Effects against saliva-derived biofilms were quantified using confocal microscopy and differential viable counting. Inhibition of biofilm formation was evaluated by pre-treating hydroxyapatite coupons with mouthrinses prior to inoculation. Otherwise-identical controls without CPC (control rinse and control fluoride rinse, CR and CFR, respectively), were included throughout. Compared to the controls, TFR and TR demonstrated significant antimicrobial effects in the redox assays, by viable counts (>3 log reductions) and in oral rinse samples (>1.25 log reductions, p 3 log difference, p \ud \ud Conclusions\ud \ud Sodium fluoride did not influence the antibacterial and anti-biofilm potency of CPC-containing formulations, supporting the combined use of CPC and sodium fluoride in mouthrinses to control oral bacteria and protect tooth enamel.