Fig. 4: Expression of selected metabolic and inflammatory genes from fracture tissue validates proteomic findings of altered metabolism and inflammation between successful and compromised bone healing.

a Heatmap of proteins related to the immune response and metabolism highlighting specific profiles for successful and compromised healing. Day 3, in particular, showed increased levels of immune and inflammatory proteins in tissue of compromised healing fractures. Proteins from metabolic pathways showed an increasing trend in successful healing fracture tissue at day 7, and a strong upregulation compared to compromised healing at day 14. Shown are sum over mean normalized LFQ intensities, with following color coding, red= upregulated, white= unchanged between conditions, blue= downregulated, grey= no values. n = 3–5 biological replicates (BR) per group and time-point. b Gene expression analysis of selected inflammatory and metabolic markers validate findings from protein levels. Pro-inflammatory genes like Ctgs, Tnf-alpha or Nos2, showed significant higher expression in compromised healing samples. The anti-inflammatory gene Il-10 and enzymes from central carbon metabolism (Hk2, Sdhb) showed higher expression in samples from successful healing. Relative expression to Tbp (TATA-binding protein, housekeeping gene) and d3 young, n = 3–6 individual biological replicates per group and time point, One-way ANOVA, mean±standard deviation.