Figure 1. Characterization of ANVs and THB@ANVs: (A) The
morphology of ANVs (left) and THB@ANVs (right), THB NPs in THB@ANVs were
marked out by triangle. Scale bar: 100 nm. (B) Coomassie blue–stained
SDS polyacrylamide gel after separation of 10 μg of total ADSCs cell
lysates or ANVs. (C) PL spectra of TPAH in the mixed solution of
ethanol/hexane with different hexane fractions. (D) Plots of the
relative emission intensity
(αAIE=I /I0 ) versus hexane
fraction. I 0 and I are the peak values of
fluorescence intensities of THB in ethanol and ethanol/hexane mixtures,
respectively. (E) Normalized absorption UV/vis spectra for ANVs, THB
NPs, and THB@ANVs in PBS. (F) Hydrodynamic diameters and (G) Zeta
potential values for ANVs, THB NPs, and THB@ANVs measured by DLS.
Diameters of lidluted (H) ANVs and (I) THB@ANVs were tested by NTA. (J)
DiO (green) and THB (red) co-localization within ANVs was assessed by
confocal microscopy. Scale bar=100 μm.
The healing and regeneration of skin wounds is a complex physiological
process that requires multiple tissues and cells to work together
synergistically in order to replace, repair, and reconstruct missing
tissue layers and cellular structures. Achieving this using a single
pharmaceutical component can be difficult, but the components found in
stem cells may fully meet this demand. Adipose-derived stem cells
(ADSCs), which can be easily obtained from adipose tissues, have been
shown to exert protective effects in several diseases through the
exosomes they release.[31] Therefore, we chose
ADSCs as our material source for preparing ANVs using a simple extrusion
method, with the aim of mimicking the functions of ADSC
exosomes.[16] As shown in Figure 1A (left), ANVs
display a tea cup holder-like structure with a size of 100-200 nm, as
observed by transmission electron microscopy (TEM). This structure is
similar to that of natural exosomes, indicating that the preparation of
ANVs was successful. The nanoparticle tracking analysis (NTA) in Figure
2I shows that the size of ANVs is 168.3±4.5 nm, with a yield of 1.99 ×
1010/106 cells. Additionally, using
the BCA detection method, the protein concentration of nanovesicles is
approximately 201 μ g/1010. In Figure 1B, the protein
profiles of ANVs match closely with the total proteins found within
whole cells, suggesting that characteristic proteins derived from ADSCs
are well-retained in ANVs.