Table 1. Compositional analysis of OMVs from P. gingivalis, T. denticola and T. forsythia.
Parametera | P. gingivalis | T. denticola | T. forsythia |
---|---|---|---|
OMVs/mL cultureb | 4.98 ± 0.77 x 1012 | 5.54 ± 1.47 x 1011 | 2.00 ± 0.65 x 1012 |
size rangec | 37–315 nm (121 nm) | 18–247 nm (75 nm) | 59–397 nm (158 nm) |
OMVs/106 nm2 cell surface aread | 936 ± 137 | 158 ± 37 | 437 ± 90 |
Protein Contente ng/108 OMVs | 38.8 ± 4.59 | 17.3 ± 2.47 | 45.4 ± 20.5 |
LPS/LOS Contentf ng/108 OMVs | 17.5 ± 5.21 | 0.11 ± 0.04 | 12.9 ± 0.64 |
Lipo-protein/peptide Contentg ng/108 OMVs | 16.2 ± 3.39 | 7.35 ± 0.39 | 14.8 ± 0.23 |
Peptidoglycan Contenth ng/108 OMVs | 2.97 ± 0.48 | 1.60 ± 0.35 | 5.21 ± 1.77 |
DNA Contenti pg/108 OMVs | 11.7 ± 3.28 | 7.44 ± 5.86 | 32.6 ± 6.74 |
RNA Contentj pg/108 OMVs | 5.11 ± 1.00 | 1.12 ± 0.50 | 4.42 ± 1.78 |
a All data are presented as mean ± standard deviation of 5 biological replicates.
b OMVs labelled with green fluorescent dye PHK-67 were counted using an Apogee A50-Micro Flow Cytometer in samples taken from late exponential cultures of each bacterium.
c Size range of OMVs was determined using Dynamic Light Scattering on a Malvern high performance particle sizer. Modal diameter in parenthesis.
d Total cell surface area was calculated using whole bacterial cell counts per mL of culture determined with a Cell Lab Quanta SC Flow Cytometer and the cellular dimensions for each species [34]
e Protein concentration determined by Qubit assay.
f Biologically active LPS was measured using HEK-Blue TLR4 Cells and a standard curve of E. coli LPS.
g Biologically active lipoproteins were measured using HEK-Blue TLR2 Cells and a standard curve of Pam3CSK4.
h Peptidoglycan was determined using a Wako SLP reagent set.
i DNA was measured using a Qubit dsDNA HS assay kit.
j RNA was measured using a Qubit RNA assay kit.