Fig. 8
ADMSCyoung-EVs, but not ADMSCold−EVs, alleviated cellular senescence and promoted M2 polarization in macrophages treated with IL-1β. a The uptake assay of ADMSCyoung-EVs and ADMSCold-EVs in cultured macrophages. The cell membranes (labeled with DIO, green), EVs (labeled with DII, red) and nuclei (DAPI, blue) are listed. The uptaken EVs per cell were analyzed. Scale bar = 100 μm and n = 4. b Western blotting and quantification of NAMPT in the control, IL-1β, IL-1β + ADMSCyoung-EVs and IL-1β + ADMSCold-EVs macrophage groups, n = 3. c Relative NAD + content in the control, IL-1β, IL-1β + ADMSCyoung-EV and IL-1β + ADMSCold-EV macrophage groups, n = 4. d Immunofluorescence staining of p16INK4A (green), p21CIP1 (red) and nuclear (DAPI, blue) in the control, IL-1β, IL-1β + ADMSCyoung-EVs and IL-1β + ADMSCold-EVs macrophage groups. Scale bar = 200 μm and n = 6. e The rate of M2 macrophages in the control, IL-1β, IL-1β + ADMSCyoung-EVs and IL-1β + ADMSCold-EVs macrophage groups, n = 4. f The relative expression of M1 markers, including CD86, iNOS, IL-6 and TLR4, and M2 markers, including Arg-1, Fizz-1, Ym-1 and CD206, in the control, IL-1β, IL-1β + ADMSCyoung-EV and IL-1β + ADMSCold-EV macrophage groups, n = 3. (*: control vs. IL-1β treatment; #: IL-1β treatment vs. IL-1β + ADMSCyoung-EVs treatment; &: IL-1β + ADMSCyoung-EVs vs. IL-1β + ADMSCold-EVs treatment groups, P < 0.05; *: P < 0.05; **: P < 0.01 and ***: P < 0.001)