Abstract
Immature dendritic cells (IDc), ‘dexosomes’, are promising natural nanomaterials for cancer diagnose and therapy. Dexosomes were isolated purely from small-scale-up production by using t25-cell-culture flasks. Total RNA was measured as 1.43 ± 0.33 ng/106 cell. Despite the fact that they possessed a surface that is highly abundant in protein, this did not become a significant effect on the DOX loading amount. Ultrasonication was used for doxorubicin (DOX) loading into the IDc dexosomes. In accordance with the literature, three candidate DOX formulations were designed as IC50 values; dExoIII, 1.8 µg/mL, dExoII, 1.2 µg/mL, and dExoI, 0.6 µg/mL, respectively. Formulations were evaluated by MTT test against highly metastatic A549 (CCL-185; ATTC) cell line. Confocal images of unloaded (naïve) were obtained by CellMaskTM membrane staining before DOX loading. Although, dexosome membranes were highly durable subsequent to ultrasonication, it was observed that dexosomes could not be stable above 70 °C during the SEM-image analyses. dExoIII displayed sustained release profile. It was found that dynamic light scattering (DLS) and nanoparticle tracking analysis (NTA) results were in good agreement with each other. Zeta potentials of loaded dexosomes have approximately between −15 to −20 mV; and, their sizes are 150 nm even after ultrasonication. IDcJAWSII dexosomes can be able to be utilized as the “BioNanoMaterial” after DOX loading via ultrasonication technique.
Highlights
Exosomes are natural nanoparticles and have some outstanding features; such as being approximately40–100 nm in size, and they are composed of different types of varied levels of adhesive proteins and amphiphilic lipid molecules in their structure after the production from various types of cells [1].Especially, Immature dendritic cells (IDc) dexosomes have versatile cellular components, such as cell penetration peptides and antigens that can direct the delivery of their cargo via both vertical and horizontal transfer [2,3]
ExoFreePBS, ExoFreeWater, and ExoFreeFBS were obtained by ultracentrifugation in phosphate-buffered saline (PBS), double-distilled water, and fetal bovine serum (FBS)
These results suggested that DOX loaded dexosomes are highly favorable cytotoxic bionanomaterial against to A549 cancer cells
Summary
IDc dexosomes have versatile cellular components, such as cell penetration peptides and antigens that can direct the delivery of their cargo (siRNA, miRNA, snRNA, or snoRNA) via both vertical and horizontal transfer [2,3] These naturally occurring events represent all of the cellular traffic which takes place even after modification of the outer surface of exosomes by genetic alteration [4,5]. IDc dexosomes, which do not have a lot of mature specific antigens because they are naïve and fresh structures, have close interaction with surface of cancer cells and can be internalized without causing an immune response [6] Unlike mature dexosomes, they do not contain any immune system-stimulating molecules. Mouse primary bone marrow-derived dendritic cells gently isolated from JAWSII (CRL-1194; ATCC)
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