Journal: Frontiers in Microbiology

Article Title: Anti-Biofilm Activity of a Self-Aggregating Peptide against Streptococcus mutans

doi: 10.3389/fmicb.2017.00488

Figure Lengend Snippet: Anti-biofilm effect of peptide P1 in microplate assay and on sHA discs. (A) Peptide P1 reduces biofilm formation by Streptococcus mutans and oral streptococci in microplate assay. Cultures were incubated with peptide P1 and control peptide sP1 at 0.1 mg/ml final concentration. Biofilm biomass in microplate wells was stained with crystal violet (images below graph). Results are combined from three replicate experiments, and error bars indicate standard error of the mean. ** p < 0.01, **** p < 0.0001, Student’s t -test. (B) Determination of minimum biofilm inhibitory concentration of peptide P1 against S. mutans . Biofilm inhibitory activity of P1 was measured by testing decreasing peptide concentrations in a microplate assay. Results were combined from three experiments, with biofilm biomass normalized to a percentage of the control. (C) Viability of S. mutans in the presence of peptide P1. Total S. mutans biomass (combined planktonic and biofilm cells from well) of cultures grown overnight in Brain Heart Infusion + 1% sucrose (BHI-S) without peptide or with 0.1 mg/ml P1 or sP1 was collected, diluted and plated, and cfu/ml was quantified for each condition. (D) Peptide P1 reduces Streptococcus mutans biofilm formation on saliva-coated hydroxyapatite (sHA) discs. Biofilm cfu/sHA disc showing combined results from three replicate experiments (total n = 9 discs/treatment). Peptide P1 and control peptide sP1 were used at 0.1 mg/ml final concentration. Error bars show standard error of the mean. * p < 0.05, using Student’s t -test. (E) Custom-made wire holder holding the hydroxyapatite disc vertically in the well of a 96-well plate.

Article Snippet: Streptococci bacteria ( Streptococcus mutans ATCC 25175, Streptococcus oralis ATCC 9811, and Streptococcus salivarius ATCC 13419), Gram-negative bacteria ( Alcaligenes faecalis ATCC 8750, Enterobacter cloacae ATCC 23355, and Salmonella typhimurium ATCC 29629), and Gram-positive bacterial strains ( ) were obtained from Yale University or Presque Isle Cultures (Erie, PA, USA).

Techniques: Incubation, Control, Concentration Assay, Staining, Activity Assay

Journal: Advanced Materials Interfaces

Article Title: Multilayered Adsorption of Commensal Microflora on Implant Surfaces: an Unconventional and Innovative Method to Prevent Bacterial Infections Associated with Biomaterials

doi: 10.1002/admi.202101410

Figure Lengend Snippet: Figure 1. Schematic representation of coating procedures. Two methods were pursued to coat S. oralis on titanium implants. A) S. oralis cultures were added as thin layers (50 µL each) on titanium discs placed directly on a hot plate adjusted to 75 °C, and a multilayered coating developed. B) S. oralis cultures were killed at 75 °C, and then titanium was incubated with these cultures at 37 °C for 5 d resulting in the formation of the coating.

Article Snippet: Growth and Cultivation of Bacteria: Streptococcus oralis (ATCC 9811, American Type Culture Collection, Manassas, USA), Streptococcus salivarius (DSM 20067, German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany), Actinomyces naeslundii (DSM 43013), Veillonella dispar (DSM 20735), Porphyromonas gingivalis (DSM 20709), and Treponema denticola (DSM 14 222) were routinely stored as glycerol stocks at –80 °C.

Techniques: Incubation

Journal: Advanced Materials Interfaces

Article Title: Multilayered Adsorption of Commensal Microflora on Implant Surfaces: an Unconventional and Innovative Method to Prevent Bacterial Infections Associated with Biomaterials

doi: 10.1002/admi.202101410

Figure Lengend Snippet: Figure 2. Macroscopic and CLSM images of uncoated and coated tita- nium. Upper part: titanium implants without coating (uncoated), after S. oralis coating (S. o.-coated), and coated implants after 48 h of immer- sion in cell culture medium (DMEM) (S. o.-coated (48 h)). Lower part: CLSM images of coated titanium before (S. o.-coated) and after 48 h of immersion in cell culture medium (S. o.-coated (48 h)). The yellow/ orange color indicates dead bacteria. Macroscopic and CLSM images of S. oralis coated implants (S.o.-coated), S. oralis coated implants after 5 d of shaking at 300 rpm (S. o.-coated (5 d)), S. oralis coated implants after shaking and brushing (S. o.-coated (b)). Coating thickness measured on S. o.-coated implants, S. o.-coated implants after 5 d of constant shaking and S. o.-coated implants after 20 times of brushing.***p < 0.0005.

Article Snippet: Growth and Cultivation of Bacteria: Streptococcus oralis (ATCC 9811, American Type Culture Collection, Manassas, USA), Streptococcus salivarius (DSM 20067, German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany), Actinomyces naeslundii (DSM 43013), Veillonella dispar (DSM 20735), Porphyromonas gingivalis (DSM 20709), and Treponema denticola (DSM 14 222) were routinely stored as glycerol stocks at –80 °C.

Techniques: Cell Culture, Bacteria

Journal: Advanced Materials Interfaces

Article Title: Multilayered Adsorption of Commensal Microflora on Implant Surfaces: an Unconventional and Innovative Method to Prevent Bacterial Infections Associated with Biomaterials

doi: 10.1002/admi.202101410

Figure Lengend Snippet: Figure 3. S. oralis coated implants antagonize the growth of bacterial bio- films. Typical CLSM images (top view and cross section) of biofilm growth on uncoated titanium surfaces, as indicated by green fluorescence of S. oralis, V. dispar, P. gingivalis, and T. denticola (left side, top to bottom). S. o.-coated titanium surfaces completely inhibit biofilms from the same pathogens (right side, top to bottom).

Article Snippet: Growth and Cultivation of Bacteria: Streptococcus oralis (ATCC 9811, American Type Culture Collection, Manassas, USA), Streptococcus salivarius (DSM 20067, German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany), Actinomyces naeslundii (DSM 43013), Veillonella dispar (DSM 20735), Porphyromonas gingivalis (DSM 20709), and Treponema denticola (DSM 14 222) were routinely stored as glycerol stocks at –80 °C.

Techniques: Fluorescence

Journal: Advanced Materials Interfaces

Article Title: Multilayered Adsorption of Commensal Microflora on Implant Surfaces: an Unconventional and Innovative Method to Prevent Bacterial Infections Associated with Biomaterials

doi: 10.1002/admi.202101410

Figure Lengend Snippet: Figure 5. S. o.-coated titanium implants resist multispecies biofilms in a flow chamber system. 3D-reconstruction of LIVE/DEAD stained multispe- cies biofilm growing on uncoated titanium (uncoated) and S.o.-coated titanium that was resistant to multispecies biofilm (S. o.-coated). Box plots of bacterial surface coverage quantified from uncoated and S.o.-coated titanium (bacterial surface coverage). FISH staining of multispecies biofilm (mul- tispecies biofilm) from uncoated implant showing four different bacterial species on the right side as; S. oralis, A. naeslundii, V. dispar, and P. gingivalis.

Article Snippet: Growth and Cultivation of Bacteria: Streptococcus oralis (ATCC 9811, American Type Culture Collection, Manassas, USA), Streptococcus salivarius (DSM 20067, German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany), Actinomyces naeslundii (DSM 43013), Veillonella dispar (DSM 20735), Porphyromonas gingivalis (DSM 20709), and Treponema denticola (DSM 14 222) were routinely stored as glycerol stocks at –80 °C.

Techniques: Staining

Journal: Advanced Materials Interfaces

Article Title: Multilayered Adsorption of Commensal Microflora on Implant Surfaces: an Unconventional and Innovative Method to Prevent Bacterial Infections Associated with Biomaterials

doi: 10.1002/admi.202101410

Figure Lengend Snippet: Figure 8. Antibiofilm properties of S. oralis coats are associated with the inhibition of bacterial adhesion. Typical force–distance curve showing A) S. oralis and C) P. gingivalis adhesion forces on S. o.-coated (gray) and uncoated titanium (black). Box plot of maximum adhesion force measured for B) S. oralis and D) P. gingivalis on uncoated (empty box) and S. o.-coated titanium (gray box).

Article Snippet: Growth and Cultivation of Bacteria: Streptococcus oralis (ATCC 9811, American Type Culture Collection, Manassas, USA), Streptococcus salivarius (DSM 20067, German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany), Actinomyces naeslundii (DSM 43013), Veillonella dispar (DSM 20735), Porphyromonas gingivalis (DSM 20709), and Treponema denticola (DSM 14 222) were routinely stored as glycerol stocks at –80 °C.

Techniques: Inhibition

Journal: Advanced Materials Interfaces

Article Title: Multilayered Adsorption of Commensal Microflora on Implant Surfaces: an Unconventional and Innovative Method to Prevent Bacterial Infections Associated with Biomaterials

doi: 10.1002/admi.202101410

Figure Lengend Snippet: Figure 7. Sole application of S. oralis led to the formation of stable and biofilm repellent coats. Typical CLSM images (top view and cross section) of P. gingivalis biofilms (green fluorescence) on uncoated, S. oralis coated (S.o.-coated) and S. salivarius coated (S.s.-coated) titanium surfaces, as well as P. gingivalis grown at the interface of S. oralis coated or uncoated titanium (Partly S.o.-coated), as well as P. gingivalis on implants coated with mixture of S. salivarius and S. oralis (S.o.-S.s.-coated). The coatings are visible as yellow/orange fluorescence of dead bacteria. White arrows indicate P. gingivalis biofilm on exposed surfaces of detached coating.

Article Snippet: Growth and Cultivation of Bacteria: Streptococcus oralis (ATCC 9811, American Type Culture Collection, Manassas, USA), Streptococcus salivarius (DSM 20067, German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany), Actinomyces naeslundii (DSM 43013), Veillonella dispar (DSM 20735), Porphyromonas gingivalis (DSM 20709), and Treponema denticola (DSM 14 222) were routinely stored as glycerol stocks at –80 °C.

Techniques: Fluorescence, Bacteria

Journal: Advanced Materials Interfaces

Article Title: Multilayered Adsorption of Commensal Microflora on Implant Surfaces: an Unconventional and Innovative Method to Prevent Bacterial Infections Associated with Biomaterials

doi: 10.1002/admi.202101410

Figure Lengend Snippet: Figure 9. Biocompatibility of S. oralis coated titanium with human gin- gival fibroblasts (HGFs). Human gingival fibroblasts were seeded on S. oralis coated and uncoated titanium samples and metabolic activity was analyzed after 24, 48, and 72 h. Graph bars show the mean ± standard deviation of gingival fibroblasts viability on uncoated titanium (white bars) and S. o.-coated titanium (gray bars) at the indicated time points. Human gingival fibroblasts on uncoated and S. oralis coated titanium after 72 h of cultivation (S. o.-coated a to c).

Article Snippet: Growth and Cultivation of Bacteria: Streptococcus oralis (ATCC 9811, American Type Culture Collection, Manassas, USA), Streptococcus salivarius (DSM 20067, German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany), Actinomyces naeslundii (DSM 43013), Veillonella dispar (DSM 20735), Porphyromonas gingivalis (DSM 20709), and Treponema denticola (DSM 14 222) were routinely stored as glycerol stocks at –80 °C.

Techniques: Activity Assay, Standard Deviation