Antimicrobial efficiency of mouthrinses versus and in combination with different photodynamic therapies on periodontal pathogens in an experimental study.

BACKGROUND AND OBJECTIVE: In the therapy of destructive periodontal disease, chemical antimicrobial agents and increasingly photodynamic therapy (PDT) play an important adjunctive role to standard mechanical anti-infective treatment procedures. However, both antiseptic methods have their shortcomings in terms of eliminating periodontal pathogens. The aim of the study was to compare the antibacterial efficacy of different antiseptic mouthrinses, of a conventional and a new, modified PDTplus as well as of the different antiseptic mouthrinses combined with either the conventional or the modified PDTplus against periopathogens. MATERIAL AND METHODS: Six representative periodontitis-associated bacterial strains were grown for 24 h under anaerobic conditions. After mixing the individual cell pellets they were exposed to 10 different antiseptic mouthrinse formulations: chlorhexidine (0.2%, 0.06%, CHX); CHX + cetylpyridinium chloride (each 0.05%); sodium hypochlorite (0.05%); polyhexanide (0.04%, PHMB1; 0.1%, PHMB2); octenidine dihydrochloride (0.1%); fluoride (250 ppm); essential oils; povidone iodine (10%); and saline (0.9%, NaCl) as control. Furthermore, the bacteria were treated with conventional PDT based on light-emitting diodes and a new modified photodisinfection combining photosensitizer with hydrogen peroxide to PDTplus also based on light-emitting diodes. In addition to the single treatments, a combined application of antiseptic exposure followed by use of PDT or PDTplus was performed. The microbial viability was characterized by analyzing colony growth and fluorescence-based vitality proportions. RESULTS: Nearly all mouthrinses caused a statistically significant growth inhibition. The most effective antiseptics, CHX (0.2%), CHX/cetylpyridinium chloride and octenidine dihydrochloride, inhibited bacterial growth completely. Conventional PDT resulted in moderate reduction of colony growth. The modified PDTplus achieved maximum antimicrobial effect. The combination of antiseptic exposure and PDT against periopathogens predominantly increased antibacterial efficacy compared to the single applications. The mouthrinse containing essential oil seemed to interfere with PDT. CONCLUSION: A combination therapy of preceding chemotherapeutical exposure and subsequent photodisinfection may be a more effective and promising antibacterial treatment than single applications of the antiseptic methods. The modified PDTplus using oxygen-enriched toluidine showed a superior antibacterial effect on periodontal pathogens to conventional PDT and to the majority of the investigated mouthrinses.

Antimicrobial effectiveness of intracanal medicaments on Enterococcus faecalis: chlorhexidine versus octenidine.

AIM: To determine the viability of Enterococcus faecalis in infected human root dentine in vitro after exposure to root canal medicaments based on chlorhexidine and octenidine. METHODOLOGY: Human root segments (n = 40) were infected with E. faecalis for 8 weeks. Root dentine samples (rd) collected at week 4 served as individual baseline values. At week 8, the root segments were randomly divided into four test groups (n = 10 each) for the placement of one of the following medicaments in the root canals: calcium hydroxide paste (CH), chlorhexidine gel (CHX-gel) (5.0%), chlorhexidine/gutta-percha points (CHX-GP) (active points(®) ; Roeko, Langenau, Germany) and octenidine gel (OCT-gel) (5.0%) followed by incubation for 4 weeks. The effect on E. faecalis viability was assessed by two fluorescent dyes (syto 9/propidium iodide) to determine the 'proportion of viable bacteria' (PVB%) and number of 'colony-forming units' (CFU). Mean values and 95% confidence intervals (CI) were calculated for PVB% and log CFU, and the difference between groups was established. RESULTS: Viable and dead bacterial cells were detected in all 'rd' samples at weeks 4 and 8. The treatment with CHX-gel, CHX-GP and OCT-gel resulted in significantly lower PVB% values with 15.4%, 3.5% and 0%, respectively. No growth (CFU) was recorded for these samples at week 12. When medicated by CH, the PVB% was increased without a corresponding change in CFUs. CONCLUSIONS: In contrast to calcium hydroxide, both CHX - and octenidine-based intracanal medicaments were effective in decreasing the viability of E. faecalis. OCT showed the most favourable results and may have potential as an endodontic medicament.

Deposition and retention of vital and dead Streptococcus sanguinis cells on glass surfaces in a flow-chamber system.

The proportion of vital as compared with dead Streptococcus sanguinis cells attached to glass surfaces was monitored and related to varying proportions of planktonic vital as compared with dead Strep. sanguinis cells. In a flow chamber with six parallel-mounted glass plates, Strep. sanguinis was suspended in pretreated sterile human saliva. Deposition of Strep. sanguinis took place, with a proportion of vital sanguinis streptococci in saliva (%VSs) of 90%, 45% or 22.5%. After exposure times of 30, 60, 90, 120 and 240 min, adherent microorganisms were labelled with two fluorescence stains to differentiate between vital and dead bacteria. Proportions of vital attached streptococci (%VSa) were determined microscopically. Dead bacteria were detected on all glass plates. The %VSa at 30 min and 60 min was significantly lower than the baseline %VSs. During the course of a single run the %VSa frequently increased after either 30, 60 or 90 min without exceeding the %VSs at 4 h. %VSs was the only variable exerting a significant effect on %VSa at 30 and 60 min. It is suggested that during the initial events of microbial attachment the dead rather than vital Strep. sanguinis cells attach preferably to solid surfaces.

Octenidine in root canal and dentine disinfection ex vivo.

AIM: The aim of the present study was to investigate the antimicrobial activity of octenidine on Enterococcus faecalis ATCC 29212 in a dentine block model. METHODOLOGY: Fifty-six root segments of extracted human teeth were infected with E. faecalis for 4 weeks. Octenidine-phenoxyethanol gel (1 : 1) was applied for different timing: 1 min, 10 min, 7 days and in a different formula (1 : 3) for 10 min. Three samples were chosen for the group with placebo gel and for the group without infection (negative control). Dentine samples were collected, and the total count of bacteria and colony-forming units were determined. In addition, for controls and the 10 min group with 1 : 1 gel, the proportion of viable bacteria (PVB) was assessed. RESULTS: Octenidine was particularly effective after incubation periods of 10 min and 7 days. The mean PVB decreased significantly from 57.2% to 5.7% after 10 min application. After 7 days, only one of 10 samples showed positive culture. CONCLUSION: The present study showed the effectiveness of octenidine against E. faecalis in dentine disinfection. Further laboratory and clinical studies are required.

A synergistic chlorhexidine/chitosan combination for improved antiplaque strategies.

BACKGROUND: The minor efficacy of chlorhexidine (CHX) on other cariogenic bacteria than mutans streptococci such as Streptococcus sanguinis may contribute to uneffective antiplaque strategies. METHODS AND RESULTS: In addition to CHX (0.1%) as positive control and saline as negative control, two chitosan derivatives (0.2%) and their CHX combinations were applied to planktonic and attached sanguinis streptococci for 2 min. In a preclinical biofilm model, the bacteria suspended in human sterile saliva were allowed to attach to human enamel slides for 60 min under flow conditions mimicking human salivation. The efficacy of the test agents on streptococci was screened by the following parameters: vitality status, colony-forming units (CFU)/ml and cell density on enamel. The first combination reduced the bacterial vitality to approximately 0% and yielded a strong CFU reduction of 2-3 log(10) units, much stronger than CHX alone. Furthermore, the first chitosan derivative showed a significant decrease of the surface coverage with these treated streptococci after attachment to enamel. CONCLUSIONS: Based on these results, a new CHX formulation would be beneficial unifying the bioadhesive properties of chitosan with the antibacterial activity of CHX synergistically resulting in a superior antiplaque effect than CHX alone.

The ability of direct fluorescence-based, two-colour assays to detect different physiological states of oral streptococci.

AIMS: To investigate the ability of six fluorescent-based, two-colour viability assays to detect different physiological growth stages of two oral streptococci species. METHODS AND RESULTS: The growth of Streptococcus sanguinis and Strep. mutans from 0 to 73 h culture periods was monitored by cell labelling with six mixtures of fluorescent stains, in addition to the growth parameters optical density (O.D.), log values of the total cell counts (log BC ml(-1)) and of the colony-forming units (log cfu ml(-1)). CONCLUSION: In comparison with the corresponding cfu values as control, the vitality proportions determined by the Syto 9/PI test best reflected the dynamic growth pattern of both test strains. The direct fluorescent-based, two-colour assay Syto 9/PI provides valuable information about microbial viability stages. SIGNIFICANCE AND IMPACT OF THE STUDY: The detection of viable but non-culturable bacteria requires more precise direct methods such as the fluorescent staining technique presented here, in addition to the classical plate count method.

Bactericidal effect of delmopinol on attached and planktonic Streptococcus sanguinis cells.

The aim of this investigation was to determine the antibacterial effect of varying concentrations of delmopinol-HCl on attached as well as on planktonic Streptooccus sanguinis cells in vitro. In addition, a possible antiadhesive effect on attached micro-organisms was to be investigated. S. sanguinis cells were allowed to attach to glass surfaces. These as well as planktonic cells were exposed to delmopinol-HCI in concentrations ranging from 0.2% to 0.00005% for 2 min. The percentage of vital bacteria was calculated by means of a fluorescence staining method. Total counts of attached bacteria were performed to determine any possible detaching effect by the delmopinol-HCl. The CFU were determined for the planktonic bacteria. Attached as well as planktonic bacteria showed a marked decrease in vitality following exposure to 0.2% delmopinol-HCl. After exposure to 0.05% this was only the case with the attached microorganisms. The total number of attached bacteria was not reduced by the delmopinol treatment. During initial dental biofilm formation, delmopinol-HCl causes a bactericidal effect when applied in concentrations of 0.05% and higher.