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.