Fig. 1

Regulatory effects of EVs from different sources on cardiomyocytes and endothelial cells. (A) PKH67-labeled exosomes were used to investigate the cellular uptake of exosomes after 24 h of co-culture, and immunofluorescence was employed for detection. Legend: To investigate the cellular uptake of exosomes from different sources, PKH67 fluorescent dye was used to label the exosomes. After a 24-hour co-culture with HUVEC or AC16 cells, the intracellular fluorescence signals were observed using immunofluorescence microscopy to assess the efficiency of exosome uptake. The figure shows the uptake of exosomes from different sources, including the negative control group (no exosomes added) and the positive control group (PKH67-labeled exosomes added). Scale bar: 10 μm. (B) The cell proliferation ability of EVs derived from different sources on HUVEC and AC16 cells was assessed using CCK8 assay. Legend: The CCK8 assay was used to evaluate the impact of EVs from different sources on the proliferation of HUVEC and AC16 cells. The results are expressed as absorbance values at 450 nm, reflecting the degree of cell proliferation. (C) EdU assay was performed to evaluate the cell proliferation ability of EVs from various sources on HUVEC and AC16 cells. Legend: The EdU assay was used to assess the impact of EVs from different sources on the proliferation of HUVEC and AC16 cells. In this assay, EdU is incorporated into newly synthesized DNA during cell replication, and the proportion of EdU-positive cells is observed under a fluorescence microscope to reflect cell proliferation. Scale bar: 100 μm. (D) Colony-formation assay was performed to evaluate the cell proliferation ability of EVs from various sources on HUVEC and AC16 cells. Legend: The colony-formation assay was used to evaluate the impact of EVs from different sources on the proliferation of HUVEC and AC16 cells. Cells were seeded in culture dishes, treated with different sources of EVs, and cultured for 10–14 days before counting the number of colonies formed. Scale bar: 100 μm. (E) Flow cytometry analysis was conducted to determine the apoptosis rate in different treatment groups. Legend: Flow cytometry was used to analyze the apoptosis rate in different treatment groups. Annexin V-FITC/PI double staining was used to detect apoptosis, showing the proportions of early apoptotic (Annexin V + PI-), late apoptotic (Annexin V + PI+), and necrotic cells (Annexin V- PI+). (F) The impact of EVs on the migratory capacity of HUVEC cells was evaluated through a wound healing assay. Legend: The wound healing assay was used to evaluate the impact of EVs from different sources on the migratory capacity of HUVEC cells. A scratch was made in a monolayer of cells, and different sources of EVs were added. Photos of the scratch healing process were taken at 0 h and 24 h. (G) Angiogenesis assays were conducted to assess the effects of AMI-EVs and N-EVs on tube formation capability. Legend: Angiogenesis assays were used to assess the effects of AMI-EVs and N-EVs on the tube formation capability of HUVEC cells. Cells were seeded on Matrigel, treated with different sources of EVs, and cultured for 6 h before observing and photographing the tube formation. Scale bar: 100 μm. Statistical significance was determined by comparing with PBS group (*P < 0.05), as well as AMI-EVs group (#P < 0.05)