Killing activity of LFchimera on periodontopathic bacteria and multispecies oral biofilm formation in vitro

ABSTRACT Lactoferrin (LF) is an iron-binding antimicrobial protein present in saliva and gingival crevicular fluids, and it is possibly associated with host defense against oral pathogens, including periodontopathic bacteria. In the present study, we evaluated the in vitro effects of LF-related agents on the growth and biofilm formation of two periodontopathic bacteria, Porphyromonas gingivalis and Prevotella intermedia, which reside as biofilms in the subgingival plaque. The planktonic growth of P. gingivalis and P. intermedia was suppressed for up to 5 h by incubation with ≥130 μg/ml of human LF (hLF), iron-free and iron-saturated bovine LF (apo-bLF and holo-bLF, respectively), and ≥6 μg/ml of bLF-derived antimicrobial peptide lactoferricin B (LFcin B); but those effects were weak after 8 h. The biofilm formation of P. gingivalis and P. intermedia over 24 h was effectively inhibited by lower concentrations (≥8 μg/ml) of various iron-bound forms (the apo, native, and holo forms) of bLF and hLF but not LFcin B. A preformed biofilm of P. gingivalis and P. intermedia was also reduced by incubation with various iron-bound bLFs, hLF, and LFcin B for 5 h. In an examination of the effectiveness of native bLF when it was used in combination with four antibiotics, it was found that treatment with ciprofloxacin, clarithromycin, and minocycline in combination with native bLF for 24 h reduced the amount of a preformed biofilm of P. gingivalis compared with the level of reduction achieved with each agent alone. These results demonstrate the antibiofilm activity of LF with lower iron dependency against P. gingivalis and P. intermedia and the potential usefulness of LF for the prevention and treatment of periodontal diseases and as adjunct therapy for periodontal diseases.

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Identification of Streptococcus sanguinis Genes Required for Biofilm Formation and Examination of Their Role in Endocarditis Virulence

Infection and Immunity ◽

10.1128/iai.00338-08 ◽

2008 ◽

Vol 76 (6) ◽

pp. 2551-2559 ◽

Cited By ~ 52

Author(s):

Xiuchun Ge ◽

Todd Kitten ◽

Zhenming Chen ◽

Sehmi P. Lee ◽

Cindy L. Munro ◽

...

Keyword(s):

Infective Endocarditis ◽

Dental Plaque ◽

Biofilm Formation ◽

Bacterial Colonization ◽

Abundant Species ◽

Oral Biofilm ◽

Streptococcus Sanguinis ◽

Streptococcal Species

ABSTRACT Streptococcus sanguinis is one of the pioneers in the bacterial colonization of teeth and is one of the most abundant species in the oral biofilm called dental plaque. S. sanguinis is also the most common viridans group streptococcal species implicated in infective endocarditis. To investigate the association of biofilm and endocarditis, we established a biofilm assay and examined biofilm formation with a signature-tagged mutagenesis library of S. sanguinis. Four genes that have not previously been associated with biofilm formation in any other bacterium, purB, purL, thrB, and pyrE, were putatively identified as contributing to in vitro biofilm formation in S. sanguinis. By examining 800 mutants for attenuation in the rabbit endocarditis model and for reduction in biofilm formation in vitro, we found some mutants that were both biofilm defective and attenuated for endocarditis. However, we also identified mutants with only reduced biofilm formation or with only attenuation in the endocarditis model. This result indicates that the ability to form biofilms in vitro is not associated with endocarditis virulence in vivo in S. sanguinis.

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Influence of topography and hydrophilicity on initial oral biofilm formation on microstructured titanium surfaces in vitro

Clinical Oral Implants Research ◽

10.1111/j.1600-0501.2011.02184.x ◽

2011 ◽

Vol 23 (3) ◽

pp. 301-307 ◽

Cited By ~ 56

Author(s):

A. Almaguer-Flores ◽

R. Olivares-Navarrete ◽

M. Wieland ◽

L. A. Ximénez-Fyvie ◽

Z. Schwartz ◽

...

Keyword(s):

Biofilm Formation ◽

Oral Biofilm ◽

Titanium Surfaces

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Experimental Models of Oral Biofilms Developed on Inert Substrates: A Review of the Literature

BioMed Research International ◽

10.1155/2016/7461047 ◽

2016 ◽

Vol 2016 ◽

pp. 1-8 ◽

Cited By ~ 12

Author(s):

Lopez-Nguyen Darrene ◽

Badet Cecile

Keyword(s):

Biofilm Formation ◽

Bacterial Species ◽

Experimental Models ◽

Pathogenic Microorganisms ◽

Review Of The Literature ◽

Complex Environment ◽

Oral Biofilm ◽

Mucosal Surfaces ◽

Oral Biofilms

The oral ecosystem is a very complex environment where more than 700 different bacterial species can be found. Most of them are organized in biofilm on dental and mucosal surfaces. Studying this community is important because a rupture in stability can lead to the preeminence of pathogenic microorganisms, causing dental decay, gingivitis, or periodontitis. The multitude of species complicates biofilm analysis so its reproduction, collection, and counting are very delicate. The development of experimental models of dental biofilms was therefore essential and multiplein vitrodesigns have emerged, each of them especially adapted to observing biofilm formation of specific bacteria within specific environments. The aim of this review is to analyze oral biofilm models.

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Plant-based oral care product exhibits antibacterial effects on different stages of oral multispecies biofilm development in vitro

BMC Oral Health ◽

10.1186/s12903-021-01504-4 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Nadine Kommerein ◽

Almut Johanna Weigel ◽

Meike Stiesch ◽

Katharina Doll

Keyword(s):

Species Distribution ◽

Biofilm Formation ◽

Antibacterial Effect ◽

Oral Care ◽

Oral Biofilm ◽

Care Product ◽

Planktonic Bacteria ◽

Multispecies Biofilm ◽

Biofilm Development

Abstract Background Excessive biofilm formation on surfaces in the oral cavity is amongst the main reasons for severe infection development like periodontitis and peri-implantitis. Mechanical biofilm removal as well as the use of adjuvant antiseptics supports the prevention of pathogenic biofilm formation. Recently, the antibacterial effect of the oral care product REPHA-OS®, based on medicinal plant extracts and essential oils, has been demonstrated on oral pathogens grown on agar plates. In the present study, the effectiveness of the product on medical relevant oral biofilm development should be demonstrated for the first time. Methods An established in vitro oral multispecies biofilm, composed of Streptococcus oralis, Actinomyces naeslundii, Veillonella dispar and Porphyromonas gingivalis, was used to analyze the antibacterial effect of different REPHA-OS® concentrations on planktonic bacteria, biofilm formation and mature biofilms. It was quantified using metabolic activity assays and live/dead fluorescence staining combined with three-dimensional confocal laser-scanning microscopy. Additionally, effects on species distribution inside the biofilm were assessed by means of quantitative real-time PCR. Results REPHA-OS® showed statistically significant antimicrobial effects on all stages of biofilm development: a minimal inhibitory concentration of 5% could be detected for both, for planktonic bacteria and for biofilm formation. Interestingly, only a slightly higher concentration of 10% was necessary to completely kill all bacteria in mature biofilms also. In contrast, an influence on the biofilm matrix or the species distribution could not be observed. The effect could be attributed to the herbal ingredients, not to the contained ethanol. Conclusion The strong antibacterial effect of REPHA-OS® on different stages of oral biofilm development strengthens its application as an alternative adjuvant in oral care therapies.

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Laser microtextured titanium implant surfaces reduce in vitro and in situ oral biofilm formation

PLoS ONE ◽

10.1371/journal.pone.0202262 ◽

2018 ◽

Vol 13 (9) ◽

pp. e0202262 ◽

Cited By ~ 9

Author(s):

Andrei C. Ionescu ◽

Eugenio Brambilla ◽

Francesco Azzola ◽

Marco Ottobelli ◽

Gaia Pellegrini ◽

...

Keyword(s):

Biofilm Formation ◽

Titanium Implant ◽

Oral Biofilm

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In Vitro Oral Biofilm Formation on Triclosan-Coated Sutures in the Absence and Presence of Additional Antiplaque Treatment

Journal of Oral and Maxillofacial Surgery ◽

10.1016/j.joms.2010.02.030 ◽

2011 ◽

Vol 69 (4) ◽

pp. 980-985 ◽

Cited By ~ 6

Author(s):

Sebastiaan Venema ◽

Frank Abbas ◽

Betsy van de Belt-Gritter ◽

Henny C. van der Mei ◽

Henk J. Busscher ◽

...

Keyword(s):

Biofilm Formation ◽

Oral Biofilm

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In vitro Characterization of Biofilm Formation in Prevotella Species

Frontiers in Oral Health ◽

10.3389/froh.2021.724194 ◽

2021 ◽

Vol 2 ◽

Author(s):

Shurooq Zakariya Albaghdadi ◽

Jenan Bader Altaher ◽

Hana Drobiova ◽

Radhika G. Bhardwaj ◽

Maribasappa Karched

Keyword(s):

Biofilm Formation ◽

Biochemical Characterization ◽

Dnase I ◽

Oral Biofilm ◽

Biofilm Inhibition ◽

Biofilm Growth ◽

In Vitro Characterization ◽

Biofilm Detachment ◽

Green Fluorescent

Background: Periodontitis, a chronic inflammatory oral infection is the outcome of disturbances in the homeostasis of the oral biofilm microbiota. A number of studies have found the occurrence of Prevotella species in elevated levels in periodontitis compared to healthy subjects. Even though different aspects of Prevotella as part of oral biofilm have been studied, in vitro biofilms formed by these species have not been characterized systematically. The objective of this study was to characterize biofilms formed by several Prevotella species and further to assess biofilm inhibition and detachment of preformed biofilms.Methods: Biofilms were grown in 24-well plates containing brucella broth in anaerobic conditions for 3 days, and were quantified using crystal violet staining. Images of SYTO 9 Green fluorescent stained biofilms were captured using confocal microscopy. Biofilm inhibition and detachment by proteinase and DNase I was tested. The biochemical characterization included quantification of proteins and DNA in the biofilms and biofilm-supernatants.Results:Prevotella loescheii, Prevotella oralis and Prevotella nigrescens showed highest biofilm formation. P. nigrescens formed significantly higher amounts of biofilms than P. loescheii (P = 0.005) and P. oralis (P = 0.0013). Inhibition of biofilm formation was significant only in the case of P. oralis when treated with proteinase (P = 0.037), whereas with DNase I treatment, the inhibition was not significant (P = 0.531). Overall, proteinase was more effective in biofilm detachment than DNase I. Protein and DNA content were higher in biofilm than the supernatant with the highest amounts found in P. nigrescens biofilm and supernatants. P. oralis biofilms appeared to secrete large amounts of proteins extracellularly into the biofilm-supernatants.Conclusion: Significant differences among Prevotella species to form biofilms may imply their variable abilities to get integrated into oral biofilm communities. Of the species that were able to grow as biofilms, DNase I and proteinase inhibited the biofilm growth or were able to cause biofilm detachment.

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