Fig. 2: AB-EVs favor adipogenesis rather than osteogenesis of BMSCs and augment osteogenic transdifferentiation of VSMCs.

a,b, ARS or ORO staining of BMSCs treated with solvent, YB-EVs, or AB-EVs under osteogenic or adipogenic induction (a) and quantification of the percentages of ARS⁺ (red) and ORO⁺ (red) areas (b). Scale bar: 50 μm. n = 6 biologically independent cells per group. c–g, ARS staining (c), quantification of the percentage of ARS⁺ areas (red; d), qRT-PCR analysis of SM22α and αSMA (e), RUNX2 and COL1A1 (f) expression, and ALP activity (g) in VSMCs treated with solvent, YB-EVs, or AB-EVs under osteogenic induction. Scale bar: 50 μm. n = 6 biologically independent cells per group. h Quantification of the percentages of ARS⁺ and ORO⁺ areas in BMSCs treated with solvent, YB-OCY-EVs, or AB-OCY-EVs under osteogenic or adipogenic induction. n = 3 biologically independent cells per group. i Quantification of the percentages of ARS⁺ areas in VSMCs treated with solvent, YB-OCY-EVs, or AB-OCY-EVs under osteogenic induction. n = 3 biologically independent cells per group. j–k, ARS and ORO staining (j) and quantification of the percentages of ARS⁺ (red) and ORO⁺ (red) areas (k) in BMSCs with different treatments under osteogenic or adipogenic induction. OC: osteoclasts; CM: conditioned media. Scale bar: 50 μm. n = 6 biologically independent cells per group. l–m, ARS staining (l) and quantification of the percentage of ARS⁺ areas (red; m) in VSMCs with different treatments under osteogenic induction. Scale bar: 50 μm. n = 6 biologically independent per group. Experiments in a–d and h–m were repeated independently three times with similar results. The illustrated results represented one of the three independent experiments. Experiments in e–g were performed with six biological replicates per group without independent repetition. Data were presented as mean ± SD. Statistical significance was determined by one-way ANOVA with Bonferroni post hoc test. Source data are provided as a Source Data file.