Extraction and Evaluation of Outer Membrane Vesicles from Two
Important Gut Microbiota Members, Bacteroides fragilis and
Ahmadi Badi S, Moshiri A, Ettehad Marvasti F, Mojtahedzadeh M, Kazemi V, Siadat SD. Extraction and evaluation of outer membrane vesicles from two important gut microbiota members, bacteroides fragilis and bacteroides thetaiotaomicron. Cell J. 2020; 22(3): 344-349. doi: 10.22074/cellj.2020.6499.
The gastrointestinal tract (GI) is colonized by a complex microbial community of gut microbiota.
Bacteroides spp. have significant roles in gut microbiota and they host interactions by various mechanisms,
including outer membrane vesicle (OMVs) production. In the present study, we extracted and assessed
Materials and Methods
In this experimental study, OMVs extraction was performed using multiple centrifugations and tris-ethylenediaminetetraacetic acid (EDTA)-sodium deoxycholate buffers. Morphology, diameter, protein content, profile, and lipopolysaccharide (LPS) concentrations of the OMVs were assessed by scanning electron microscopy (SEM), nanodrop, Bradford assay, sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), and the Limulus Amoebocyte Lysate (LAL) test, respectively. Zeta potential (ζ-P) was also assessed. The viability effect of OMVs was assessed by the 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay in Caco-2 cells.
Spherical OMVs with diameters of 30-110 nm were produced. The OMVs had different protein profiles. The
LPS concentrations of the
The endotoxin concentrations of the spherical OMVs from