Lipopolysaccharide Impaired the Functional Activity of Endothelial Colony-Forming Cells

Abstract

Background: Recent studies have shown that endothelial progenitor cells (EPCs) contribute to lung repair after lipopolysaccharide (LPS)-induced lung injury and infusion of LPS decreased early EPCs in human peripheral blood. However, the effects of LPS on endothelial colony-forming cells (ECFCs) remain to be determined. Objective: To investigate possible effects of LPS on the functional activity of ECFCs. Methods: ECFCs were isolated from human umbilical cord blood and characterized. ECFCs at passages 3-5 were treated for 24 h with either LPS or vehicle control. Their viability, migration and in vitro vasculogenesis activity were assayed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, modified Boyden chamber and in vitro angiogenesis assays, respectively. ECFC adhesion was assessed by replating cells on fibronectin-coated dishes and subsequent counting of adherent cells. Results: Incubation with LPS dose-dependently inhibited the viable, migratory, adhesive and in vitro vasculogenesis capacity of ECFCs. Conclusion: LPS impaired the functional activity of ECFCs.

Toll-like receptor 4 is expressed and functional in late endothelial progenitor cells

“…The authors previously demonstrated that LPS impaired the viability, migration, adhesion and in vitro vasculogenesis capacity of late EPCs (13). On this basis, the present study revealed that inhibition of TLR4 signaling can reverse this impairment of functional activity in late EPCs, which suggested that TLR4 signaling pathways may be involved in the LPS-induced dysfunction of late EPCs.…”

“…The present study was approved by the ethics committee of Shantou University Medical College (Guangdong, China). Late EPCs were isolated and cultured according to previously MIN YU 1,2* , CANTIAN WANG 1* , GUONING ZENG 1 , LIHONG ZHOU 1 , TINGTING CHEN 1 , XUERUI TAN 1 and YANPING WANG 1 described techniques (3,13,(15)(16)(17). Mononuclear cells (MNCs) were isolated from previously obtained umbilical cord blood from the First Affiliated Hospital of Shantou University Medical College (13) by Ficoll density gradient centrifugation (TDL-5-A, Shanghai Anting Scientific Instrument Factory, Shanghai, China) at 1,000 x g for 20 min at room temperature and cultured on 6-well plates with endothelial cell growth medium (EGM)-2 (Lonza Group, Ltd., Basel, Switzerland Membranes were blocked for 1 h at room temperature with 5% non-fat milk in TBS containing 0.05% Tween-20 (TBST) and incubated with the polyclonal rabbit anti-TLR4 antibody (1:500) at 4˚C overnight, followed by 1 h incubation at room temperature with a horseradish peroxidase-conjugated secondary goat polyclonal anti-rabbit antibody (cat.…”

“…The functional activity of late EPCs were determined according to previously described methods (13,(15)(16)(17).…”

“…Unlike early EPCs, late EPCs have not been studied in great detail. The authors previously demonstrated that lipopolysaccharide (LPS) inhibited the viability, migration, adhesion and in vitro angiogenesis of late EPCs (13). However, the mechanisms by which LPS induces the impairment of late EPC functional activity remain unknown.…”

A Fermented Whole Grain Prevents Lipopolysaccharides‐Induced Dysfunction in Human Endothelial Progenitor Cells

“…Anyway, in our study, adhesion capacity of “early” EPCs was strengthened by treatment with LG. Moving from “early” EPCs to endothelial colony-forming cells (ECFCs), also called “late” EPCs, it is worthy to mention that incubation with LPS inhibited the viable, migratory, adhesive, and in vitro vasculogenesis capacity of ECFCs isolated from human umbilical cord blood [48]. …”