Interkingdom biofilm of Streptococcus pyogenes and Candida albicans: establishment of an in vitro model and dose-response validation of antimicrobials.

Although Streptococcus pyogenes and Candida albicans may colonize tonsillar tissues, the interaction between them in mixed biofilms has been poorly explored. This study established an interkingdom biofilm model of S. pyogenes and C. albicans and verified the dose-response validation of antimicrobials. Biofilms were formed on microplates, in the presence or absence of a conditioning layer of human saliva, using Brain Heart Infusion (BHI) broth or artificial saliva (AS) as a culture medium, and with variations in the microorganism inoculation sequence. Biofilms grown in AS showed higher mass than those grown in BHI broth, and an opposite trend was observed for metabolism. The number of S. pyogenes colonies was lower in AS. Amoxicillin and nystatin showed dose-dependent effects. The inoculation of the two species at the same time, without prior exposure to saliva, and using BHI broth would be the model of choice for future studies assessing the effects of antimicrobials on dual S. pyogenes/C. albicans biofilms.

Buffering Capacity and Effects of Sodium Hexametaphosphate Nanoparticles and Fluoride on the Inorganic Components of Cariogenic-Related Biofilms In Vitro.

Despite the remarkable effects of sodium hexametaphosphate nanoparticles (HMPnano) on dental enamel de-/re-mineralization processes, information on the effects of these nanoparticles on biofilms is scarce. This study assessed the effects of HMPnano, with or without fluoride (F), on the inorganic components and pH of Streptococcus mutans and Candida albicans dual-species biofilms. Solutions containing conventional/micro-sized HMP (HMPmicro) or HMPnano were prepared at 0.5% and 1%, with or without 1100 ppm F. A 1100 ppm F solution and pure artificial saliva were tested as positive and negative controls, respectively. The biofilms were treated three times and had their pH analyzed, and the concentrations of F, calcium, phosphorus, and HMP in the biofilm biomass and fluid were determined. In another set of experiments, after the last treatment, the biofilms were exposed to a 20% sucrose solution, and the biofilm pH and inorganic components were evaluated. The 1% HMPnano solution with F led to the highest biofilm pH, even after exposure to sucrose. The 1% HMPnano solution without F led to significantly higher phosphorus concentrations in comparison to all other groups. It can be concluded that 1% HMPnano and F influenced the biofilm pH, besides affecting most of the inorganic components of the dual-species biofilms.

In vitro antimicrobial effects of chitosan on microcosm biofilms of oral candidiasis.

OBJECTIVE: This study assessed the effects of chitosan (CS) on microcosm biofilms derived from saliva of patients with Candida-associated denture stomatitis. METHODS: Five removable denture wearers with denture stomatitis were included in the study. The minimum inhibitory concentration (MIC) of CS against clinical isolates of Candida albicans was determined according to the broth microdilution method. Pooled saliva from the donors was used as an inoculum for the formation of biofilms, which were developed during 72 h on acrylic surfaces in the Amsterdam Active Attachment model. The biofilms were then treated with different concentrations of CS, and the antibiofilm effects were evaluated through the quantification of colony-forming units (CFUs), total biomass (TB), metabolic activity (MA), lactic acid production (LAP), and cell viability (by confocal laser scanning microscopy). Chlorhexidine, miconazole, and nystatin were tested as positive controls, while the negative control (NC) was the untreated biofilm. Data were analyzed by 1-way ANOVA and Fischer LSD's post hoc test (α=0.05). RESULTS: MIC values of CS ranged from 500 to 800 µg/mL. For CFUs, 2500 µg/mL CS was the most effective treatment in reducing total anaerobes, mutans streptococci, and Lactobacillus spp., significantly differing from the controls. For C. albicans CFUs, CS and positive controls did not differ from each other but led to significant reductions compared to NC. Regarding TB, MA, LAP, and cell viability, 2500 µg/mL CS promoted the greatest reductions compared to NC. CONCLUSION: CS has similar or superior effects to conventional active principles on important parameters of oral candidiasis microcosm biofilms. CLINICAL RELEVANCE: The antibiofilm effects of CS show that this compound has great potential to improve the clinical condition of denture stomatitis patients, and formulations containing this natural polymer could be useful for controlling oral candidiasis.