Fig. 3: iMASSAGE produces large quantities of therapeutic EV.

a Schematic diagram of iMASSAGE-induced EVs generation under wireless stimulation. b Concentration of EVs released from normal culture (w/o iMASSAGE) and iMASSAGE treatment (with iMASSAGE, 1 W, 2 min), measured with NTA. Macrophage RAW264.7 were used. c Relative EV amounts detected by flow cytometry. Using CD63 expression as an internal measure of total vesicle counts. Upper: representative SEM images of EVs immuno-captured on anti-CD63 labeled microbeads. More EVs were covered on captured beads for iMASSAGE treatment medium. Scale bar: 400 nm. Down: flow cytometry analysis of bead-bound EVs, after labeling with anti-CD9 conjugated dye. d Representative TEM images of EVs collected from normal culture and iMASSAGE treatment, respectively. Scale bar: 200 nm. e SDS-PAGE protein analysis of EV collected from different culture conditions. f The abundance of miRNA-125b and miR-149 present in normal culture (w/o iMASSAGE) and iMASSAGE treatment-derived EVs, separately. All data were made relative to that of w/o iMASSAGE (n = 3 independent experiments). g Viability and morphology (upper right) of embedded macrophage cells under multiple days of iMASSAGE treatments with one-day intervals (Calcein AM stained). Scale bar: 10 μm (up) and 50 μm (down). h Relative concentrations of EVs produced at different days (n = 3 independent experiments). iMASSAGE stimulation (3 T and 1 W, blue dot) was employed on days 1, 3, and 5, without stimulation on days 2 and 5 (black dot). All data were normalized against normally secreted EVs (without stimulation group), obtained without iMASSAGE treatment. All data are presented as mean ± s.d. Source data are provided as a Source Data file.