Bacterial Adhesion of TESPSA and Citric Acid on Different Titanium Surfaces Substrate Roughness: An In Vitro Study with a Multispecies Oral Biofilm Model.

This in vitro study analyzed the influence of substrate roughness on biofilm adhesion and cellular viability over triethoxysilylpropyl succinic anhydride silane (TESPSA)- and citric acid (CA)-coated surfaces at 12 and 24 h, respectively. A multispecies biofilm composed of S. oralis, A. naslundii, V. parvula, F. nucleatum, P. intermedia, P. gingivalis, P. endodontalis and F. alocis was developed over titanium discs grouped depending on their roughness (low, medium, high) and antibacterial coating (low-TESPSA, medium-TESPSA, high-TESPSA, and CA). The biofilm was quantified by means of quantitative polymerase chain reaction (PCR) and viability PCR and assessed through confocal laser scanning microscope (CLSM). Quantitative PCR revealed no significant differences in bacterial adhesion and biofilm mortality. CA was the surface with the lowest bacterial counts and highest mortality at 12 and 24 h, respectively, while high harbored the highest amount of biofilm at 24 h. By CLSM, CA presented significant amounts of dead cells compared to medium-TESPSA and high-TESPSA. A significantly greater volume of dead cells was found at 12 h in low-TESPSA compared to medium-TESPSA, while CA also presented significant amounts of dead cells compared to medium-TESPSA and high-TESPSA. With regard to the live/dead ratio, low-TESPSA presented a significantly higher ratio at 12 h compared to medium-TESPSA and high-TESPSA. Similarly, CA exhibited a significantly higher live/dead ratio compared to medium-TESPSA and high-TESPSA at 12 h. This multispecies in vitro biofilm did not evidence clear antiadhesive and bactericidal differences between surfaces, although a tendency to reduce adhesion and increase antibacterial effect was observed in the low-TESPSA and CA.

The quorum quenching enzyme Aii20J modifies in vitro periodontal biofilm formation.

Introduction: Recent studies have revealed the presence of N-acyl-homoserine lactones (AHLs) quorum sensing (QS) signals in the oral environment. Yet, their role in oral biofilm development remains scarcely investigated. The use of quorum quenching (QQ) strategies targeting AHLs has been described as efficient for the control of pathogenic biofilms. Here, we evaluate the use of a highly active AHL-targeting QQ enzyme, Aii20J, to modulate oral biofilm formation in vitro. Methods: The effect of the QQ enzyme was studied in in vitro multispecies biofilms generated from oral samples taken from healthy donors and patients with periodontal disease. Subgingival samples were used as inocula, aiming to select members of the microbiota of the periodontal pocket niche in the in vitro biofilms. Biofilm formation abilities and microbial composition were studied upon treating the biofilms with the QQ enzyme Aii20J. Results and Discussion: The addition of the enzyme resulted in significant biofilm mass reductions in 30 - 60% of the subgingival-derived biofilms, although standard AHLs could not be found in the supernatants of the cultured biofilms. Changes in biofilm mass were not accompanied by significant alterations of bacterial relative abundance at the genus level. The investigation of 125 oral supragingival metagenomes and a synthetic subgingival metagenome revealed a surprisingly high abundance and broad distribution of homologous of the AHL synthase HdtS and several protein families of AHL receptors, as well as an enormous presence of QQ enzymes, pointing to the existence of an intricate signaling network in oral biofilms that has been so far unreported, and should be further investigated. Together, our findings support the use of Aii20J to modulate polymicrobial biofilm formation without changing the microbiome structure of the biofilm. Results in this study suggest that AHLs or AHL-like molecules affect oral biofilm formation, encouraging the application of QQ strategies for oral health improvement, and reinforcing the importance of personalized approaches to oral biofilm control.

Bacterial decontamination of toothbrushes by immersion in a mouthwash containing 0.05% chlorhexidine and 0.05% cetylpyridinium chloride: A randomized controlled trial.

OBJECTIVE: Toothbrushes are colonized by microorganisms, implying a risk of infection. That risk can be reduced by decreasing the microbial contamination of the filaments. Therefore, this study aimed to determine the antiseptic efficacy of a 0.05% chlorhexidine + 0.05% cetylpyridinium chloride mouthwash on toothbrushes. METHODS: A total of twelve toothbrushes used three times/day for 14 days by orally and systemically healthy people were randomly split into two groups, and their heads were immersed for 2 h in PBS (control) or Perio·Aid Active Control (treatment). The microorganisms were recovered, and their number was calculated by culture, quantitative PCR, and viability PCR. Statistical differences were first assessed with a two-way mixed ANOVA and subsequently with Student's t-test. RESULTS: The results showed no statistical differences in the total number of cells for the treatment (mean ± CI95% of 7.27 ± 1.09 log10 bacteria/ml) and the control (7.62 ± 0.64 log10 bacteria/ml) groups, but a significantly lower number of live cells in the treatment group (4.58 ± 0.61 log10 viable bacteria/ml and 2.15 ± 1.42 log10 cfu/ml) than in the control group (6.49 ± 1.39 log10 viable bacteria/ml and 5.04 ± 0.93 log10 cfu/ml). CONCLUSIONS: Based on our findings, sanitization of toothbrushes with this mouthwash reduces the number of live microorganisms adhered to the filaments. Such decrease of the bacterial load could include bacteria from the oral cavity, from the environment, and from nearby toothbrushes since the quantification was not limited to any bacterial taxon.

Development and viability of biofilms grown on experimental abutments mimicking dental implants: An in vivo model

Background: To determine whether an experimental abutment mimicking the macro- and microstructure of a dental implant is a suitable method for recovering biofilm, and to describe the features of biofilms formed around such abutments on healthy implants. Material and Methods: Experimental abutments were used in 15 patients without peri-implant diseases. After 14 days' absence of dental hygiene in this area, the abutments were retrieved and analyzed through confocal laser scanning microscopy and scanning electron microscopy. The biofilm formation on the surface of the first 5 abut-ments was determined by a fluorescence-staining method using SYTO9 nucleic acid stain. In order to study the biofilm's coverage and vitality, 10 additional abutments were assessed using live & dead bacterial viability. Descriptive and bivariate analyses of the data were performed. Results: A global plaque coverage of the abutments was observed in all cases. The submucosal area of the abutment was mostly covered with biofilm (over 21%). Moreover, significant differences between supra- and subgingival locations were detected. Conclusions: This in vivo experimental model allows detailed observation of the extensive plaque growth found on exposed experimental abutments mimicking dental implants when hygiene measures are absent. The biofilm cover-age is significantly higher in the supragingival zone than in the subgingival portion

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