Journal: Journal of Periodontal & Implant Science

Article Title: A periodontitis-associated multispecies model of an oral biofilm

doi: 10.5051/jpis.2014.44.2.79

Figure Lengend Snippet: Biofilm growth curves. P. gingivalis : Porphyromonas gingivalis , F. nucleatum : Fusobacterium nucleatum , S. gordonii : Streptococcus gordonii , A. actinomycetemcomitans : Aggregatibacter actinomycetemcomitans , CFU: colony forming unit.

Article Snippet: S. gordonii KN1, F. nucleatum ATCC 23726, A. actinomycetemcomitans ATCC 33384, and P. gingivalis ATCC 33277 were obtained from the culture collection of the Department of Oral Microbiology, Gangneung-Wonju National University Dental College.

Techniques:

Journal: Journal of Periodontal & Implant Science

Article Title: A periodontitis-associated multispecies model of an oral biofilm

doi: 10.5051/jpis.2014.44.2.79

Figure Lengend Snippet: Confocal micrographs represent a two-dimensional maximum projection of the biofilms after 24 hours, from the biofilm surface (left image) to the deepest layer of the biofilm (right image) (×400): (A) Porphyromonas gingivalis , (B) Fusobacterium nucleatum , (C) Streptococcus gordonii , (D) Aggregatibacter actinomycetemcomitans , and (E) multispecies bacteria.

Article Snippet: S. gordonii KN1, F. nucleatum ATCC 23726, A. actinomycetemcomitans ATCC 33384, and P. gingivalis ATCC 33277 were obtained from the culture collection of the Department of Oral Microbiology, Gangneung-Wonju National University Dental College.

Techniques: Bacteria

Journal: Journal of Periodontal & Implant Science

Article Title: A periodontitis-associated multispecies model of an oral biofilm

doi: 10.5051/jpis.2014.44.2.79

Figure Lengend Snippet: Representative scanning electron microscopy images of the biofilms after 24 hours (×30,000): (A) Porphyromonas gingivalis , (B) Fusobacterium nucleatum , (C) Streptococcus gordonii , (D) Aggregatibacter actinomycetemcomitans , and (E) multispecies bacteria. Bars=1 µm.

Article Snippet: S. gordonii KN1, F. nucleatum ATCC 23726, A. actinomycetemcomitans ATCC 33384, and P. gingivalis ATCC 33277 were obtained from the culture collection of the Department of Oral Microbiology, Gangneung-Wonju National University Dental College.

Techniques: Electron Microscopy, Bacteria

Journal: Frontiers in Immunology

Article Title: Fusobacterium nucleatum Extracellular Vesicles Modulate Gut Epithelial Cell Innate Immunity via FomA and TLR2

doi: 10.3389/fimmu.2020.583644

Figure Lengend Snippet: Characterization of EVs derived from F. nucleatum . (A) density gradient fractions of total protein profile stained with Coomassie Blue. (B) EVs fractions identified by far-western blot using a FomA specific biotinylated peptide derived from statherin protein, highlighted by the red rectangle. (C) EVs fractions 2 to 8 imaged by TEM; scale bar, 100 nm. Yellow arrows give examples of EVs structures (D) EVs count using NTA methods in 7 extraction batches, normalized by the volume of culture. (E) Size (nm) distribution of EVs in 7 distinct cultures and extractions, black lines represent individual EVs extraction, red line shows the mean distribution of 6 different purifications. (F) Protein concentration in each purification as determined by the BCA method and normalized by the initial volume of culture, biomass measured by optical density (OD) at 600 nm.

Article Snippet: Fusobacterium nucleatum subsp nucleatum DSM 15643 - ATCC 23726 was cultured in DSMZ M104 medium depleted from meat extract and resazurin in Hungate tubes under CO 2 or in bottles in an anaerobic cabinet (82% N 2 and 18% CO 2 ) for 24 h. Bacterial viability status was assessed before each EVs extraction using LIVE/DEAD BacLight Bacterial Viability Kit (Invitrogen).

Techniques: Derivative Assay, Staining, Far Western Blot, Extraction, Protein Concentration, Purification

Journal: Frontiers in Immunology

Article Title: Fusobacterium nucleatum Extracellular Vesicles Modulate Gut Epithelial Cell Innate Immunity via FomA and TLR2

doi: 10.3389/fimmu.2020.583644

Figure Lengend Snippet: Impact of EVs on intestinal epithelial cells and innate immunity. (A) Epithelial resistance monitored in Caco-2 differentiated cells exposed to 10 µg.ml −1 EV for 5 h or Blank EV (Ω.cm²). Data are represented as the average of three experiments ± SD. Exact p-value of EVs vs Blank EV were 1 h: 0.110; 2 h: 0.093; 3 h: 0.104; 4 h: 0.188; 5 h: 0.199 (T test). (B) NF-κB activation level in Caco-2, T84 and HT-29 NF-κB reporter cell lines after incubation of 24 h with different EVs concentrations. Caco-2 and HT-29 p-values were not significant. T84 p-value from left to right: 0.0357, 0.0090, and 0.0095. Data are expressed as median ± quartiles of fold change toward unstimulated cells. (C) Secreted IL-8 measured by ELISA in HCTT16, HT-29 stimulated with 5µg.ml −1 EVs, Blank EV or TNF-α 10ng.ml −1 for 24 h (N = 3). (D) Secreted IL-8 measured by ELISA in T84 cells stimulated with F. nucleatum supernatant, control medium, 5 µg.ml −1 EVs, Blank EV or TNF-α 10 ng.ml −1 for 24 h (N = 4). (E) (Upper left panel) 5 µg of FITC-labeled EVs or (Lower left panel) 5 µg of Blank EVs, were incubated on T84 cells for 3 h, 100X, FITC (Green), nucleus DAPI (Blue); scale bar, 10µm.

Article Snippet: Fusobacterium nucleatum subsp nucleatum DSM 15643 - ATCC 23726 was cultured in DSMZ M104 medium depleted from meat extract and resazurin in Hungate tubes under CO 2 or in bottles in an anaerobic cabinet (82% N 2 and 18% CO 2 ) for 24 h. Bacterial viability status was assessed before each EVs extraction using LIVE/DEAD BacLight Bacterial Viability Kit (Invitrogen).

Techniques: Activation Assay, Incubation, Enzyme-linked Immunosorbent Assay, Control, Labeling