Bone Marrow Mesenchymal Stem Cell-Derived Extracellular Vesicles Carrying circ_0050205 Attenuate Intervertebral Disc Degeneration

Figure 2

BMSC-EVs stimulated the survival of NPCs and inhibit ECM degradation in NPCs by transferring circ_0050205. (a) RT-qPCR analysis of circ_0050205 expression in the NP tissues of IDD mice () and sham-operated mice (), vs. sham-operated mice. (b) RT-qPCR analysis of circ_0050205 expression in tBHP-exposed NPCs, vs. tBHP-0 μM. (c) Transfer of circ_0050205-Cy3 from BMSCs into NPCs via EVs (25 μm). (d) RT-qPCR analysis of circ_0050205 expression in tBHP-exposed NPCs cocultured with BMSC-EVs, vs. the tBHP group. (e) RT-qPCR analysis of circ_0050205 expression in EVs derived from circ_0050205-overexpressing BMSCs, vs. vector. (f) RT-qPCR analysis of circ_0050205 expression in tBHP-exposed NPCs cocultured with EVs derived from circ_0050205-overexpressing BMSCs, vs. tBHP + BMSC-EVs + vector. (g) CCK-8 assay for the viability of tBHP-exposed NPCs cocultured with EVs circ_0050205-overexpressing BMSCs, vs. tBHP + BMSC-EVs + vector. (h) EdU staining for proliferation of tBHP-exposed NPCs cocultured with EVs from circ_0050205-overexpressing BMSCs, vs. tBHP + BMSC-EVs + vector. (i) Flow cytometric analysis of apoptosis of tBHP-exposed NPCs cocultured with EVs from circ_0050205-overexpressing BMSCs, vs. tBHP + BMSC-EVs + vector. (j) Immunofluorescence detection of MMP13, ADAMTS 5, COL II, and aggrecan proteins in tBHP-exposed NPCs cocultured with EVs from circ_0050205-overexpressing BMSCs (25 μm). (k) The ROS production was examined using the DCF-DA kit in tBHP-exposed NPCs cocultured with EVs from circ_0050205-overexpressing BMSCs, vs. tBHP + BMSC-EVs + Vector. (l) Lipid peroxidation determined by C11-BODIPY581/591 in tBHP-exposed NPCs cocultured with EVs from circ_0050205-overexpressing BMSCs. (m) The total free iron in tBHP-exposed NPCs cocultured with EVs from circ_0050205-overexpressing BMSCs, vs. tBHP + BMSC-EVs + vector. . Cell experiments were conducted three times independently.

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