Comparison of the efficiency of ultrafiltration, precipitation, and ultracentrifugation methods for exosome isolation.

Extracellular vesicles (EVs) are enclosed by a lipid-bilayer membrane and secreted by all types of cells. They are classified into three groups: apoptotic bodies, microvesicles, and exosomes. Exosomes play a number of important roles in the intercellular communication and crosstalk between tissues in the body. In this study, we use three common methods based on different principles for exosome isolation, namely ultrafiltration, precipitation, and ultracentrifugation. We use field emission scanning electron microscopy (FESEM) and dynamic light scattering (DLS) analyses for characterization of exosomes. The functionality and effect of isolated exosomes on the viability of hypoxic cells was investigated by alamarBlue and Flow-cytometry. The results of the FESEM study show that the ultrafiltration method isolates vesicles with higher variability of shapes and sizes when compared to the precipitation and ultracentrifugation methods. DLS results show that mean size of exosomes isolated by ultrafiltration, precipitation, and ultracentrifugation methods are 122, 89, and 60 nm respectively. AlamarBlue analysis show that isolated exosomes increase the viability of damaged cells by 11%, 15%, and 22%, respectively. Flow-cytometry analysis of damaged cells also show that these vesicles increase the content of live cells by 9%, 15%, and 20%, respectively. This study shows that exosomes isolated by the ultracentrifugation method are characterized by smaller size and narrow size distribution. Furthermore, more homogenous particles isolated by this method show increased efficiency of the protection of hypoxic cells in comparison with the exosomes isolated by the two other methods.

Green synthesis of zinc oxide nanoparticles loaded on activated carbon prepared from walnut peel extract for the removal of Eosin Y and Erythrosine B dyes from aqueous solution: experimental approaches, kinetics models, and thermodynamic studies.

Water contamination due to release of dye containing effluents is one of the environmental problems of serious concern today. The present study investigate the green synthesis of zinc oxide nanoparticles (ZnO-NPs) doped on activated carbon (AC) prepared from walnut peel extract and to estimate its efficiency in the removal of Eosin Y (Eo-Y) and Erythrosine B (Er-B) from its aqueous solution. The synthesized AC-ZnO was identified by field emission scanning electron microscopy (FE-SEM), X-ray diffraction (XRD), and the Brunauer-Emmett-Teller. The influence of various parameters such as pH, dosage of AC-ZnO, contact time, and concentrations of Eo-Y and Er-B was also studied. The pH 3 was observed as the optimum pH while the equilibrium was noticed to reach in 30 min at dosage of 1 g/L and initial concentration 100 mg/L for Eo-Y and Er-B adsorption onto AC-ZnO. The maximum adsorption capacity of Eo-Y and Er-B onto AC-ZnO was found to be 163.9 and 144.92 mg/g (and removal efficiencies of 95.11 and 98.31 %), respectively. The process of Eo-Y and Er-B adsorption on AC-ZnO was observed to be depended on the pseudo-second-order kinetic model which indicates chemisorption processes. Langmuir adsorption isotherm model test described the removal of Eo-Y and Er-B on AC-ZnO. The thermodynamic data indicated that the adsorption was endothermic process. Also, the values, SBET and VTOTAL, for the AC-ZnO were equal to 725.65 m2/g and 0.6004 cm3/g, respectively. The results of this study exhibited that AC-ZnO was a very effective method that can be used for the removal of Eo-Y and Er-B from aqueous solutions.