Supplementary data on confocal imaging of Porphyromonas gingivalis release of polyphosphates and fibrin formation
The hypothesis was that Porphyromonas gingivalis by secreting gingipains or polyphosphates could initiate blood coagulation and thereby contribute to fibrin deposition in gingival tissue which in turn promotes chronic inflammation. Chronic inflammation in gingival tissue eventually leads to bone degradation and loss of tooth support, known as periodontitis. The results show that P. gingivalis initiate coagulation by means of gingipains as well as polyphosphates. Supplementary data on confocal imaging of P. gingivalis release of polyphosphates and fibrin formation; mages and microscopy movies.
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Planktonic suspensions and biofilm cells of P. gingivalis were prepared. The planktonic suspensions were transferred to an Ibidi® μ-slide and incubated anaerobically for two hours to allow the cells to adhere to the surface. The Ibidi μ-slide was then centrifuged at 1200g for 10min. DAPI (4’,6-Diamidino-2-Phenylindole) was added to a final concentration of 18μM (5μg/mL) and the slide was incubated in the dark for 60min. Fluorescence was excited in a spinning disk confocal microscope (Nikon Eclipse TE2000 with a CFI Plan Apokromat 60x oil lens, Nikon Corp., Tokyo, Japan). For visualization of DNA the excitation wavelength was 378nm and the emission wavelength was 456nm. Excitation at 415nm and emission at 550nm showed polyP . Images were acquired with a Photometrics Prime 95B camera using Nikon NIS-Elements software.