mRNA-seq and DEG analyses of MSCs grown in control microcryogels and OS-Microcryogels after 7 d of osteogenic induction. (A) Schematic of osteogenic induction before mRNA-seq. (B) Heatmap distribution of control microcryogels and OS-Microcryogels. (C) Volcano diagram of control microcryogels and OS-Microcryogels (2185 downregulated DEGs, 3213 upregulated DEGs). (D) The significant GO of upregulated DEGs indicated that the effects of OS- Microcryogels on the regulation of MSCs were associated with osteoblast differentiation, ossification, bone development, bone growth, and bone mineralization. (E) KEGG enrichment analyses of targeted genes indicated that the effects of OS-Microcryogels on the regulation of MSCs were associated with the PI3K-Akt, FoxO, TGF-β, MAPK, and Wnt signaling pathways. (F) GSEA of COL1A1 in GO terms of activation of MAPK activity, bone cell development, positive regulation of macrophage migration, regulation of macrophage migration, transforming growth factor-beta production and vascular associated smooth muscle contraction. (G) GSEA of RUNX2 in the GO terms of positive regulation of osteoclast differentiation, response to transforming growth factor-beta, etc.