Abstract
BACKGROUND: Nanoparticles, as small extracellular vesicles, are considered promising tools in tissue engineering and regenerative medicine. This study aimed to investigate the effects of different processing and culture condition on the quality and quantity of extracts derived from human Adipose-Mesenchymal Stem Cells (AD-MSCs). METHODS: AD-MSCs were proliferated in both the experimental and control groups. Nanoparticles were extracted from AD-MSCs-extracts and analyzed using SEM, TEM, DLS, Zeta potential, FTIR and BCA analyses. The morphological characteristics (shape, size, distribution, surface topography, and agglomeration/aggregation), structural appearance (poly-disperse intensity, colloidal particle behavior, surface charge, and stability), chemical properties (functional groups and ionic interactions) and total protein concentration were detected in the extracted nanoparticles. Additionally, the morphological characteristics, apoptosis, mitochondrial oxidoreductase activity, and migration potential of AD-MSCs in both groups were evaluated using acridine orange staining, MTT, and scratch assays. RESULTS: In the experimental group, 100% of the nanoparticles had a diameter of 112.8± 25 nm, with the most frequency of 111.4 nm. However, in the control group, 72% of nanoparticles had a diameter of 350.2±43.6 nm with the highest frequency of 339.8 nm (p≤0.05). The Z-average, Poly-disperse intensity, and electrostatic stability of nanoparticles in the control and experimental groups were 171.9 nm, 0.727 and -0.000011 cm(2) /Vs vs. 103.7 nm, 0.205 and 0.000481 cm(2) /Vs, respectively (p≤0.05). In the experimental group, Zeta potential was -61.8 mV, which is in the range of ζ >-30mV. Although, Zeta potential in the control group was -1.5 mV, which is in the range of -30 mV <ζ <30 mV (p≤0.05). Total protein concentrations in the control and experimental groups were 11 and 41%, respectively (p≤0.05). CONCLUSION: Nanoparticles derived from AD-MSCs have high therapeutic applications in tissue engineering and regenerative medicine.