Fig. 5: Evaluation of the ability of Mg²⁺-loaded microrobots to promote bone formation in vitro.

A Live/Dead staining of BMSCs after coculture with the indicated microspheres for 3 and 7 days. B CCK-8 assays of BMSCs after coculture with the indicated microspheres for 1, 3, 5, and 7 days. Data are presented as mean ± SD (n = 3 biologically independent samples, two-way ANOVA with Dunnett’s multiple comparisons test and adjustment applied). C Crystal violet staining of migrated BMSCs in the Transwell experiment. D Scratch wound healing assays of BMSCs at 0 and 36 h. E ALP staining of BMSCs after coculture with the indicated microspheres for 7 and 14 days in osteogenic-conditioned medium. F ARS of BMSCs after coculture with the indicated microspheres for 14 and 21 days in osteogenic-conditioned medium. G BMP-2, RUNX-2, and OCN immunofluorescence staining (green) of BMSCs. The cells were costained with DAPI (blue) and phalloidin (red). H Relative mRNA expression of BMP-2, RUNX-2, OCN, and ALP in BMSCs. Data are presented as mean ± SD (n = 3 biologically independent samples, one-way ANOVA with Dunnett’s multiple comparisons test and adjustment applied). I Schematic illustration of Mg²⁺-loaded microrobots promoting the proliferation, migration, and osteogenesis of BMSCs. I created in BioRender. Ding, Z. (2025) https://BioRender.com/p69t917. Source data are listed in the Source Data file.