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Effectiveness of repeated photodynamic therapy in the elimination of intracanal Enterococcus faecalis biofilm: an in vitro study.

ABSTRACT: The study aimed to investigate the effectiveness of photodynamic therapy in the elimination of intracanal Enterococcus faecalis biofilm and to analyse how a repeated light irradiation, replenishment of oxygen and photosensitiser affect the results of the photodynamic disinfecting protocol. After chemomechanical preparation, 46 single-rooted human teeth were infected with a clinical strain of E. faecalis and incubated for a week in microaerobic conditions. The experimental procedures included groups of single application of photodynamic therapy, two cycles of PDT, irrigation with 5.25% NaOCl solution and negative and positive control. The number of residing bacterial colonies in the root canals was determined based on the CFU/ml method. In the group of preparations irrigated with NaOCl, bacterial colonies were not observed. A single PDT eliminated 45% of the initial CFU/ml. Repeated PDT eradicated 95% of the intracanal bacterial biofilm. Photodynamic therapy has a high potential for the elimination of E. faecalis biofilm. There is a safe therapeutic window where photoinduced disinfection can be used as an adjuvant to conventional endodontic treatment, which remains the most effective.

SUBMITTER: Prazmo EJ

PROVIDER: S-EPMC5360838 | biostudies-literature | 2017 Apr

REPOSITORIES: biostudies-literature

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Effectiveness of repeated photodynamic therapy in the elimination of intracanal Enterococcus faecalis biofilm: an in vitro study.

Prażmo Ewa Joanna EJ Godlewska Renata Alicja RA Mielczarek Agnieszka Beata AB

Lasers in medical science 20170210 3

The study aimed to investigate the effectiveness of photodynamic therapy in the elimination of intracanal Enterococcus faecalis biofilm and to analyse how a repeated light irradiation, replenishment of oxygen and photosensitiser affect the results of the photodynamic disinfecting protocol. After chemomechanical preparation, 46 single-rooted human teeth were infected with a clinical strain of E. faecalis and incubated for a week in microaerobic conditions. The experimental procedures included gro ...[more]

PMID: 28185089

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Project description:BackgroundEnterococcus faecalis and Candida albicans are frequently found in persistent endodontic infection and could remain in dentinal tubules despite intracanal medication with calcium hydroxide (Ca(OH)2), a commonly used medication. Thus, an effective and safe antimicrobial medication against such refractory infection is necessary in endodontic retreatment, so we aimed to test the efficacy of chitosan paste against these microorganisms compared with Ca(OH)2 in root canals of extracted human teeth.MethodsThirty-six sterilized human root samples prepared from extracted premolars and upper maxillary incisors were infected with E. faecalis for 14 days, while 32 were infected with C. albicans for 48 h, for mature biofilm formation. The samples were assigned to 6 groups of intracanal medications: Group 1: no medication (negative control); Group 2: 20% Polyethylene glycol (PEG); Group 3: 20% Propylene glycol (PG); Group 4: Ca(OH)2; Group 5: Chitosan + PEG; and Group 6: Chitosan + PG. After 7 days, intracanal surface dentin was harvested using Protaper next, resuspended, serially diluted and spread on Brain-Heart-Infusion agar (for E. faecalis) and Yeast Extract-Peptone-Dextrose agar (for C. albicans) for colony count. Antimicrobial efficacy was determined as percentage of remaining colony forming unit (CFUs) relative to negative control and analyzed using One-way ANOVA and post-hoc Games-Howell test. The significance level was set at 0.05.ResultsFor E. faecalis, chitosan + PG had significantly higher antibacterial activity than Ca(OH)2 (P = 0.039). Chitosan + PEG and chitosan + PG medication significantly reduced viable bacteria compared with negative control, PEG and PG (P = 0.001, 0.003, 0.024, respectively for chitosan + PEG; P = 0.002, 0.003, 0.014, respectively for chitosan + PG). For C.albicans, chitosan + PEG and chitosan + PG were not significantly different from Ca(OH)2. However, Chitosan + PEG and chitosan + PG, but not Ca(OH)2, showed a significantly lower level of remaining CFUs compared with negative control (P = 0.013 and 0.005, respectively).ConclusionChitosan paste showed better efficacy in reducing viable E. faecalis biofilm when compared to Ca(OH)2 after 7-day intracanal medication in this in vitro root canal model. It could also significantly reduce viable C. albicans, but was not significantly different from Ca(OH)2.

Project description:Introduction: Enterococcus faecalis is a resistant bacterium which is the most abundant species in infected root canals. Photodynamic therapy (PDT) is a method for killing the bacteria with active Oxygen radicals generated in a photosensitizer when exposed to centralized light. Furthermore, as a new method of canal disinfection, a variety of irrigation activation systems have been introduced, one of which is GentleFile (GF) with rotary movements and spiral effects for antibacterial action. This study aimed to compare the effectiveness of the two mentioned methods when used with and without Sodium Hypochlorite in eliminating E. faecalis from infected root canals. Methods: Fifty-eight uniradicular teeth were randomly divided into 4 experimental groups of 14. Two specimens were selected for later scanning electron microscopy in order to screen the procedure steps. In each experimental group, 10 samples were selected to be treated with GF or PDT; 3 of them were selected as positive controls and the other one sample was chosen as a negative control. Experimental groups were as follows: (1) Irrigation activation system, (2) Irrigation activation system + sodium hypochlorite, (3) PDT, and (4) PDT+ sodium hypochlorite. The specimens were then cultured for a bacterial colony count. Results: The decrease in the bacterial count after the treatment with the irrigation activation system was 99.8% (P =0.011) and when the system was used with sodium hypochlorite, it was 100% (P =0.001). The antibacterial effect of PDT was 90.08% (P =0.011) and it was 99.7% when PDT was combined with sodium hypochlorite (P =0.011). Conclusion: All four methods can be administered as complementary methods in root canal disinfection. According to the results of disinfection in the experimental groups of current study it is concluded that integration of new technologies such as activation irrigation system or PDT in Combination with NaOCl ameliorates disinfection of root canal and can provide several advantages in the endodontic outcome.

Project description:Enterococcus faecalis is a common commensal organism and a prolific nosocomial pathogen that causes biofilm-associated infections. Numerous E. faecalis OG1RF genes required for biofilm formation have been identified, but few studies have compared genetic determinants of biofilm formation and biofilm morphology across multiple conditions. Here, we cultured transposon (Tn) libraries in CDC biofilm reactors in two different media and used Tn sequencing (TnSeq) to identify core and accessory biofilm determinants, including many genes that are poorly characterized or annotated as hypothetical. Multiple secondary assays (96-well plates, submerged Aclar discs, and MultiRep biofilm reactors) were used to validate phenotypes of new biofilm determinants. We quantified biofilm cells and used fluorescence microscopy to visualize biofilms formed by six Tn mutants identified using TnSeq and found that disrupting these genes (OG1RF_10350, prsA, tig, OG1RF_10576, OG1RF_11288, and OG1RF_11456) leads to significant time- and medium-dependent changes in biofilm architecture. Structural predictions revealed potential roles in cell wall homeostasis for OG1RF_10350 and OG1RF_11288 and signaling for OG1RF_11456. Additionally, we identified growth medium-specific hallmarks of OG1RF biofilm morphology. This study demonstrates how E. faecalis biofilm architecture is modulated by growth medium and experimental conditions and identifies multiple new genetic determinants of biofilm formation. IMPORTANCE E. faecalis is an opportunistic pathogen and a leading cause of hospital-acquired infections, in part due to its ability to form biofilms. A complete understanding of the genes required for E. faecalis biofilm formation as well as specific features of biofilm morphology related to nutrient availability and growth conditions is crucial for understanding how E. faecalis biofilm-associated infections develop and resist treatment in patients. We employed a comprehensive approach to analysis of biofilm determinants by combining TnSeq primary screens with secondary phenotypic validation using diverse biofilm assays. This enabled identification of numerous core (important under many conditions) and accessory (important under specific conditions) biofilm determinants in E. faecalis OG1RF. We found multiple genes whose disruption results in drastic changes to OG1RF biofilm morphology. These results expand our understanding of the genetic requirements for biofilm formation in E. faecalis that affect the time course of biofilm development as well as the response to specific nutritional conditions.

Dataset Information

Effectiveness of repeated photodynamic therapy in the elimination of intracanal Enterococcus faecalis biofilm: an in vitro study.

Publications

Effectiveness of repeated photodynamic therapy in the elimination of intracanal Enterococcus faecalis biofilm: an in vitro study.

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