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Ex vivo vs. in vivo antibacterial activity of two antiseptics on oral biofilm.

Prada-López I, Quintas V, Casares-De-Cal MA, Suárez-Quintanilla JA, Suárez-Quintanilla D, Tomás I - Front Microbiol (2015)

Bottom Line: Fifteen healthy volunteers wore a specific intraoral device for 48 h to form a biofilm in three glass disks.Samples were analyzed for bacterial viability with the confocal laser scanning microscope after previous staining with LIVE/DEAD® BacLight™.The method of application conditioned the antibacterial activity of the 0.2% CHX and EO solutions on the in situ oral biofilm.

View Article: PubMed Central - PubMed

Affiliation: Oral Sciences Research Group, Special Needs Unit, School of Medicine and Dentistry, University of Santiago de Compostela Santiago de Compostela, Spain.

ABSTRACT

Aim: To compare the immediate antibacterial effect of two application methods (passive immersion and active mouthwash) of two antiseptic solutions on the in situ oral biofilm.

Material and methods: A randomized observer-masked crossover study was conducted. Fifteen healthy volunteers wore a specific intraoral device for 48 h to form a biofilm in three glass disks. One of these disks was used as a baseline; another one was immersed in a solution of 0.2% Chlorhexidine (0.2% CHX), remaining the third in the device, placed in the oral cavity, during the 0.2% CHX mouthwash application. After a 2-weeks washout period, the protocol was repeated using a solution of Essential Oils (EO). Samples were analyzed for bacterial viability with the confocal laser scanning microscope after previous staining with LIVE/DEAD® BacLight™.

Results: The EO showed a better antibacterial effect compared to the 0.2% CHX after the mouthwash application (% of bacterial viability = 1.16 ± 1.00% vs. 5.08 ± 5.79%, respectively), and was more effective in all layers (p < 0.05). In the immersion, both antiseptics were significantly less effective (% of bacterial viability = 26.93 ± 13.11%, EO vs. 15.17 ± 6.14%, 0.2% CHX); in the case of EO immersion, there were no significant changes in the bacterial viability of the deepest layer in comparison with the baseline.

Conclusions: The method of application conditioned the antibacterial activity of the 0.2% CHX and EO solutions on the in situ oral biofilm. The in vivo active mouthwash was more effective than the ex vivo passive immersion in both antiseptic solutions. There was more penetration of the antiseptic inside the biofilm with an active mouthwash, especially with the EO. Trial registered in clinicaltrials.gov with the number NCT02267239. URL: https://clinicaltrials.gov/ct2/show/NCT02267239.

No MeSH data available.


Related in: MedlinePlus

Protocol of the study.
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Figure 1: Protocol of the study.

Mentions: To calculate an a priori sample size, the following statistical criteria were established: an effect size of 0.35, an alpha error of 0.05 and a statistical power of 87%. Assuming these criteria and using the repeated measures ANOVA test, a sample size of 15 subjects was required. The sample size calculation was performed using the program G*Power 3.1.5. The participants were recruited among dental students at the Faculty of Medicine and Dentistry of Santiago de Compostela (Spain), where volunteer enrolment was asked by responding to advertisements for the participation in a research study at the faculty hall. All of these volunteers were revised by the same trained clinician to ensure they fulfilled all inclusion and exclusion criteria. The volunteers chosen met the same inclusion and exclusion criteria of previous publications of our group (García-Caballero et al., 2013; Prada-López et al., 2015a; Quintas et al., 2015). The inclusion criteria were the following: being systemically healthy adult volunteers between 20 and 45 years old, who presented a good oral health status: a minimum of 24 permanent teeth with no evidence of gingivitis or periodontitis (Community Periodontal Index score = 0) (WHO, 1997) and an absence of untreated caries at the beginning of the study. The following exclusion criteria were applied: smoker or former smoker, presence of dental prostheses or orthodontic devices, antibiotic treatment or routine use of oral antiseptics in the previous 3 months, and presence of any systemic disease that could alter the production or composition of saliva. Before the start of each phase, a full mouth scaling with ultrasonic instruments and teeth polishing with rubber cup after dental disclosure was performed by the same trained clinician on all selected participants (Figure 1). Written informed consent was obtained from all participants in the study.


Ex vivo vs. in vivo antibacterial activity of two antiseptics on oral biofilm.

Prada-López I, Quintas V, Casares-De-Cal MA, Suárez-Quintanilla JA, Suárez-Quintanilla D, Tomás I - Front Microbiol (2015)

Protocol of the study.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4488754&req=5

Figure 1: Protocol of the study.
Mentions: To calculate an a priori sample size, the following statistical criteria were established: an effect size of 0.35, an alpha error of 0.05 and a statistical power of 87%. Assuming these criteria and using the repeated measures ANOVA test, a sample size of 15 subjects was required. The sample size calculation was performed using the program G*Power 3.1.5. The participants were recruited among dental students at the Faculty of Medicine and Dentistry of Santiago de Compostela (Spain), where volunteer enrolment was asked by responding to advertisements for the participation in a research study at the faculty hall. All of these volunteers were revised by the same trained clinician to ensure they fulfilled all inclusion and exclusion criteria. The volunteers chosen met the same inclusion and exclusion criteria of previous publications of our group (García-Caballero et al., 2013; Prada-López et al., 2015a; Quintas et al., 2015). The inclusion criteria were the following: being systemically healthy adult volunteers between 20 and 45 years old, who presented a good oral health status: a minimum of 24 permanent teeth with no evidence of gingivitis or periodontitis (Community Periodontal Index score = 0) (WHO, 1997) and an absence of untreated caries at the beginning of the study. The following exclusion criteria were applied: smoker or former smoker, presence of dental prostheses or orthodontic devices, antibiotic treatment or routine use of oral antiseptics in the previous 3 months, and presence of any systemic disease that could alter the production or composition of saliva. Before the start of each phase, a full mouth scaling with ultrasonic instruments and teeth polishing with rubber cup after dental disclosure was performed by the same trained clinician on all selected participants (Figure 1). Written informed consent was obtained from all participants in the study.

Bottom Line: Fifteen healthy volunteers wore a specific intraoral device for 48 h to form a biofilm in three glass disks.Samples were analyzed for bacterial viability with the confocal laser scanning microscope after previous staining with LIVE/DEAD® BacLight™.The method of application conditioned the antibacterial activity of the 0.2% CHX and EO solutions on the in situ oral biofilm.

View Article: PubMed Central - PubMed

Affiliation: Oral Sciences Research Group, Special Needs Unit, School of Medicine and Dentistry, University of Santiago de Compostela Santiago de Compostela, Spain.

ABSTRACT

Aim: To compare the immediate antibacterial effect of two application methods (passive immersion and active mouthwash) of two antiseptic solutions on the in situ oral biofilm.

Material and methods: A randomized observer-masked crossover study was conducted. Fifteen healthy volunteers wore a specific intraoral device for 48 h to form a biofilm in three glass disks. One of these disks was used as a baseline; another one was immersed in a solution of 0.2% Chlorhexidine (0.2% CHX), remaining the third in the device, placed in the oral cavity, during the 0.2% CHX mouthwash application. After a 2-weeks washout period, the protocol was repeated using a solution of Essential Oils (EO). Samples were analyzed for bacterial viability with the confocal laser scanning microscope after previous staining with LIVE/DEAD® BacLight™.

Results: The EO showed a better antibacterial effect compared to the 0.2% CHX after the mouthwash application (% of bacterial viability = 1.16 ± 1.00% vs. 5.08 ± 5.79%, respectively), and was more effective in all layers (p < 0.05). In the immersion, both antiseptics were significantly less effective (% of bacterial viability = 26.93 ± 13.11%, EO vs. 15.17 ± 6.14%, 0.2% CHX); in the case of EO immersion, there were no significant changes in the bacterial viability of the deepest layer in comparison with the baseline.

Conclusions: The method of application conditioned the antibacterial activity of the 0.2% CHX and EO solutions on the in situ oral biofilm. The in vivo active mouthwash was more effective than the ex vivo passive immersion in both antiseptic solutions. There was more penetration of the antiseptic inside the biofilm with an active mouthwash, especially with the EO. Trial registered in clinicaltrials.gov with the number NCT02267239. URL: https://clinicaltrials.gov/ct2/show/NCT02267239.

No MeSH data available.


Related in: MedlinePlus