Validation of Antibiotic Efficacy on In Vitro Subgingival Biofilms

Belibasakis, Georgios N.; Thurnheer, Thomas

doi:10.1902/jop.2013.130167

Cited by 41 publications

(49 citation statements)

References 31 publications

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“…Furthermore, S. mutans colonizes in vitro much less efficiently on streptococcal biofilms than on A. naeslundii biofilms [Wang et al, 2011]. As compared to the Zurich biofilm model of supragingival plaque and its various modifications comprising up to 11 bacterial or fungal species [Guggenheim et al, 2001;Shapiro et al, 2002;Ammann et al, 2012;Belibasakis and Thurnheer, 2014], this biofilm model is less complex in terms of species diversity but, on the other hand, allows easy and high-throughput screening while still being robust to antimicrobial challenge. Treatment with CHX 2.0% for 20 min served as a positive control leading to total eradication of biofilm bacteria below detection limit.…”

Section: Discussionmentioning

confidence: 99%

Phenalen-1-One-Mediated Antimicrobial Photodynamic Therapy and Chlorhexidine Applied to a Novel Caries Biofilm Model

Wimmer¹,

Muehler²,

Thurnheer

et al. 2018

Caries Res

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Antimicrobial photodynamic therapy (aPDT) may be useful as a supportive antimicrobial measure for caries-active subjects. In this study, the antimicrobial efficacy of aPDT with a phenalen-1-one photosensitizer was evaluated in a novel in vitro biofilm model comprising Actinomyces naeslundii, Actinomyces odontolyticus, and Streptococcus mutans and was compared to chlorhexidine. The proposed biofilm model allows high-throughput screening for antimicrobial efficacy while exhibiting a differentiated response to different antimicrobial approaches. While chlorhexidine 0.2% showed a reduction of ≈4 log₁₀ for all species, aPDT led to a more pronounced reduction of S. mutans (2.8 log₁₀) than of Actinomyces spp. (1.2 or 1.3 log₁₀). A similar effect was also observed in monospecies biofilms. Therefore, aPDT may be more effective against S. mutans than against Actinomyces spp. when in biofilms, and this antimicrobial approach merits further investigations.

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Section: Discussionmentioning

confidence: 99%

Phenalen-1-One-Mediated Antimicrobial Photodynamic Therapy and Chlorhexidine Applied to a Novel Caries Biofilm Model

Wimmer¹,

Muehler²,

Thurnheer

et al. 2018

Caries Res

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“…There are few other studies examining the effects of antibiotics on polymicrobial oral bacterial biofilms. Belibasakis and Thurnheer [18] recently reported that amoxicillin + metronidazole (1:1 ratio) at a concentration of 15 mg/L caused reductions to total cell numbers of established 10-species polymicrobial biofilms, significantly reducing P. gingivalis numbers in these biofilms after 24 h of exposure. Soares et al have recently reported that the amoxicillin + metronidazole combination significantly reduced metabolic activity of 35 subgingival bacterial species residing in complex biofilms [47].…”

Section: Discussionmentioning

confidence: 99%

“…As late colonisers in dental biofilm formation [15], the red complex bacteria ( Porphyromonas gingivalis , Treponema denticola , and Tannerella forsythia ) express synergistic virulence and pathogenicity [16,17]. Among the antibiotics used in treating periodontitis, amoxicillin in combination with metronidazole has been shown to display a strong effect in reducing numbers of non-periodontal bacteria, as well as P. gingivalis and Fusobacterium nucleatum monomicrobial biofilms in vitro [18], and it is currently proposed to be the most clinically and microbiologically advantageous adjunctive antibiotic regime in treating periodontitis [19,20]. Azithromycin, a macrolide antibiotic [21], has also gained popularity for the treatment of periodontitis [22–25].…”

Section: Introductionmentioning

confidence: 99%

Effect of azithromycin on a red complex polymicrobial biofilm

Ong

Oettinger‐Barak

Dashper

et al. 2017

Journal of Oral Microbiology

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Azithromycin has recently gained popularity for the treatment of periodontal disease, despite sparse literature supporting efficiency in treating periodontal bacterial biofilms. The aim of this study was to evaluate the effect of azithromycin on biofilms comprised of Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia in comparison to an amoxicillin and metronidazole combination. P. gingivalis W50, T. denticola ATCC35405, and T. forsythia ATCC43037 grown under anaerobic conditions at 37°C were aliquoted into 96-well flat-bottom plates in different combinations with addition of azithromycin or amoxicillin + metronidazole at various concentrations. For the biofilm assay, the plates were incubated at 37°C anaerobically for 48 h, after which the biofilms were stained with crystal violet and measured for absorbance at AU620. In this model, polymicrobial biofilms of P. gingivalis + T. denticola, P. gingivalis + T. forsythia, and T. denticola + T. forsythia were cultured. Combination of all three bacteria enhanced biofilm biomass. Azithromycin demonstrated a minimal biofilm inhibitory concentration (MBIC) of 10.6 mg/L, while the amoxicillin + metronidazole combination was more effective in inhibiting biofilm formation with a MBIC of 1.63 mg/L. Polymicrobial biofilm formation was demonstrated by combination of all three red complex bacteria. Azithromycin was ineffective in preventing biofilm formation within a clinically achievable concentration, whereas the combination of amoxicillin and metronidazole was more effective for this purpose.

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“…Clear guidelines and distinct protocols are still lacking. Therapy decisions are based on interventional studies and in vitro experiments [36–39]. Commercially available kits and PCR primers are currently used in periodontal risk assessment, in monitoring therapeutic success and as an aid in choosing appropriate antibiotics [40–48].…”

Section: Discussionmentioning

confidence: 99%

Accuracy of commercial kits and published primer pairs for the detection of periodontopathogens

et al. 2016

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ObjectivesDespite the input of microbiome research, a group of 20 bacteria continues to be the focus of periodontal diagnostics and therapy. The aim of this study was to compare three commercial kits and laboratory-developed primer pairs for effectiveness in detecting such periodontopathogens.Materials and methodsFourteen bacterial mock communities, consisting of 16 randomly assembled bacterial strains, were used as reference standard for testing kits and primers. Extracted DNA from mock communities was analyzed by PCR in-house with specific primers and forwarded for analysis to the manufacturer’s laboratory of each of the following kits: ParoCheck®Kit 20, micro-IDent®plus11, and Carpegen® Perio Diagnostik.ResultsThe kits accurately detected Fusobacterium nucleatum, Prevotella intermedia/Prevotella nigrescens, Parvimonas micra, Aggregatibacter actinomycetemcomitans, Campylobacter rectus/showae, Streptococcus mitis, Streptococcus mutans, and Veillonella parvula. The in-house primers for F.nucleatum were highly specific to subtypes of the respective periopathogen. Other primers repeatedly detected oral pathogens not present in the mock communities, indicating reduced specificity.ConclusionsThe commercial kits used in this study are reliable tools to support periodontal diagnostics. Whereas the detection profile of the kits is fixed at a general specificity level, the design of primers can be adjusted to differentiate between highly specific strains. In-house primers are more error-prone. Bacterial mock communities can be established as a reference standard for any similar testing.Clinical relevanceThe tested kits render good results with selected bacterial species. Primers appear to be less useful for routine clinical diagnostics and of limited applicability in research. Basic information about the periodontopathogens identified in this study supports clinical decision-making.

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Validation of Antibiotic Efficacy on In Vitro Subgingival Biofilms

Abstract: Antibiotics at concentrations detectable in gingival crevicular fluid do not dramatically reduce total bacterial loads in this in vitro biofilm model, but cause species-specific reductions, which may disrupt the biofilm unity.

Cited by 41 publications

References 31 publications

Phenalen-1-One-Mediated Antimicrobial Photodynamic Therapy and Chlorhexidine Applied to a Novel Caries Biofilm Model

Phenalen-1-One-Mediated Antimicrobial Photodynamic Therapy and Chlorhexidine Applied to a Novel Caries Biofilm Model

Effect of azithromycin on a red complex polymicrobial biofilm

Accuracy of commercial kits and published primer pairs for the detection of periodontopathogens

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