Fig. 5: Engineering and characterization of CPP-miR-Exo. | Nature Communications

Fig. 5: Engineering and characterization of CPP-miR-Exo.

From: Injectable ECM-mimetic dynamic hydrogels abolish ferroptosis-induced post-discectomy herniation through delivering nucleus pulposus progenitor cell-derived exosomes

Fig. 5

a Engineering procedures of CPP-miR-Exo. b Representative TEM images of miR-Exo and CPP-miR-Exo. Scale bars, 200 nm. c CLSM images illustrate the cellular uptake of CPP-miR-Exo in NPCs. miR-Exo and CPP were labeled with PKH26 and FITC, respectively. Scale bars, 10 μm. d Representative fluorescence intensity profiles of FITC compared with PKH26 in the radial direction of the white lines in (c). Size distribution (e) and ζ-potential (f) of miR-Exo and CPP-miR-Exo. n = 5 biologically independent samples for panel (f). Western blot bands (g) and quantitative analysis (h) showing the expression of exosome markers in miR-Exo and CPP-miR-Exo (n = 6 biologically independent samples). Fluorescence images (i) and quantitative analysis (j) demonstrating the time-dependent internalization of miR-Exo or CPP-miR-Exo at 50 μg/mL in NPCs (n = 5 biologically independent samples). Scale bar, 100 μm. Representative flow cytometric profiles (k–l) and quantitative analysis (m) of the uptake of miR-Exo in NPCs after 4 h of co-culture (n = 5 biologically independent samples). Comparisons were performed by two-tailed Student’s t test in panels (h, j and m). Data are presented as means ± SD. n = 3 independent experiments for panels (b, c, e). NPPC nucleus pulposus progenitor cell, NPPC-Exo NPPC-derived exosome, miR-Exo miR-221-3p overexpressed NPPC-Exo CPP, cell-penetrating peptide, CPP-miR-Exo cell-penetrating peptide modified miR-Exo. Schematic illustrations were generated using BioRender (Wang, V., 2025; accessible at: https://BioRender.com/i62o243). Source data are provided as a Source Data file.

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