Fig. 9: GaCuVan&HACHO-BSA hydrogel, administered in advance, contributed to resistance to ectopic osteomyelitis and induced trained immunity through metabolic reprogramming in rats.
A Ectopic osteomyelitis study protocols including pre-delivery of different hydrogels, establishment of ectopic MRSA-infected osteomyelitis, followed by treatments across groups (G1, Sham group; G2, PBS group; G3, Van group; G4, GaVan group; G5, GaCuVan group; G6, GaCuVan&HACHO-BSA group) and subsequent transcriptomic, immunological, histopathological, and bacteriological analyses (Created in BioRender. Chen, H. (2025) https://BioRender.com/n054ax4). B Heat map and functional classification of differentially expressed genes in bone marrow of rats with ectopic osteomyelitis pretreated with GaCuVan&HACHO-BSA hydrogel and Van (fold change ≥ 2 and q < 0.05). C GO enrichment analysis of the upregulated genes in GaCuVan&HACHO-BSA hydrogel group and Van group. D KEGG enrichment analysis of the upregulated DEGs in GaCuVan&HACHO-BSA hydrogel group and Van group. E–H GSEA analysis was performed on GO, KEGG, Reactome, and Wikipathway datasets from the rat, and the significantly enriched gene sets ranked in the top 5 by absolute NES value were selected by default for map display. Representative HIF-1α (I) and COX2 (J) immunofluorescence images of bone marrow sections on day 14 of MRSA infection with ectopic osteomyelitis and quantification of HIF-1α (K) and COX2 (L) protein. Cytokine levels of IL-17 (M), IL-6 (N), and TNF-α (O) in bone marrow from ectopic osteomyelitis rats on day 14 were detected by an ELISA kit. Cytokine levels of IL-17 (P), IL-6 (Q), and TNF-α (R) in serum from ectopic osteomyelitis rats on day 14 were measured using an ELISA kit. The data in (K–R) were expressed as the mean ± s.d. (n = 4 biologically independent samples per group), with the results shown as mean ± SEM. Statistical analysis was performed using One-way ANOVA with Tukey’s multiple comparison test. A was generated using BioRender.com, released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license.
