Exosomes Derived From Low‐Intensity Pulsed Ultrasound‐Treated Dendritic Cells Suppress Tumor Necrosis Factor–Induced Endothelial Inflammation

Abstract

Objectives Endothelial cell inflammation plays an important role in atherosclerosis. Low‐intensity pulsed ultrasonography (LIPUS) exerts an anti‐inflammatory function on endothelial cells, whereas the underlying mechanism has not been fully elucidated. Methods Bone marrow dendritic cells (BMDCs) derived from bone barrow cells were treated with LIPUS, and exosomes secreted into the supernatant were purified. The isolated exosomes were incubated with human umbilical vein endothelial cells (HUVECs) to investigate their effect on tumor necrosis factor (TNF)‐α–induced endothelial inflammation. Ultrastructure was analyzed by transmission electron microscopy. Messenger RNA levels were determined by quantitative reverse transcription polymerase chain reaction, and protein levels were analyzed by western blot. Results The isolated exosomes presented a typical exosomal size of 30 to 100 nm in diameter and expressed exosome positive markers (Alix, CD63, and TSG101) but not the exosome negative marker (Calnexin). Exosomes derived from LIPUS‐treated BMDCs were rich in miR‐16 and miR‐21, which could be engulfed by HUVECs. Pretreatment with exosomes impeded TNFα‐induced HUVEC activation and downregulated TNFα‐stimulated expression of vascular cell adhesion molecule‐1 and intercellular adhesion molecule‐1, thus preventing TNFα‐induced activation of the nuclear factor‐κB signaling pathway. Conclusion Exosomes derived from LIPUS‐treated BMDC inhibit TNFα‐induced endothelial inflammation by inhibiting the nuclear factor‐κB signaling pathway.

Dendritic cell derived exosomes loaded with immunoregulatory cargo reprogram local immune responses and inhibit degenerative bone disease in vivo

“…From these DC subtypes, EXO were purified, quantitated and correct size distribution (30–150nm) confirmed by nanoparticle tracking analysis ( Figure 1A ). Further characterization indicates predominant population of EXO, as determined by immunogold-CD63 TEM ( Figure 1B ), immunoblot for Tsg101 (component of Escort1 complex), CD81, B-actin, Alix (Escort Associated Protein) and EXO negative marker GRP94 ( Figure 1C ), as previously reported [ 43 ].…”

“…EXO donor DC subtypes including immunoregulatory DCs (regDCs), immature DCs (iDCs), and immunostimulatory (stimDCs) were generated from bone marrow derived non-adherent cells of C57BL/6 mice, and phenotype and cytokine profiles characterized as follows: regDCs:CD11c +, low MHCII+, low CD86+, low CD80 +and low CD40+ Figure 1A). Further characterization indicates predominant population of EXO, as determined by immunogold-CD63 TEM ( Figure 1B), immunoblot for Tsg101 (component of Escort1 complex), CD81, B-actin, Alix (Escort Associated Protein) and EXO negative marker GRP94 ( Figure 1C), as previously reported [43].…”

Aberrant expression profiles and bioinformatic analysis of CAF-derived exosomal miRNAs from three moderately differentiated supraglottic LSCC patients

“…2s). Previous studies have reported that exosomes are generally enriched with certain speci c marker proteins, such as CD9, CD63, CD81, TSG101 and HSP70 [23,24], with lower expression of some cellular proteins, such as cis-Golgi matrix protein GM130 [25] and endoplasmic reticulum (ER) calnexin [14,15]. Exosomes can therefore be identi ed by western blotting or ow cytometry.…”

“…2s). Western blotting revealed positive expression of CD63 and TSG101, two of the well-established surface markers for exosomes, and negative expression of calnexin, commonly used as a negative control for extracellular vesicles [14,15] (Fig. 2t).…”

Current Perspectives on Clinical Use of Exosomes as a Personalized Contrast Media and Theranostics

“…Here, doxorubicin was delivered into glioma tumors with the help of FUS, while no significant differences in physical features, ultrasound-responsiveness, or specific glioma targeting were observed [82]. Moreover, another modality of ultrasonography called low-intensity pulse ultrasonography (LIPUS) as well as its impact on exosomes was studied by Li and colleagues [83]. Bone marrow dendritic cells (BMDC) were extracted from dead C57BL/6 mice's bone marrow and part of them was treated with LIPUS.…”