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. 2025 Oct 14;56:15–32. doi: 10.1016/j.bioactmat.2025.09.041

Fig. 4.

Fig. 4

Recapitulating Bone Microenvironment-Mediated Tumor Suppression in Prostate Cancer. a) Schematic and fluorescence images showing C4-2B tumor spheroids cultured with either CPC scaffolds (control) or the BME model. b) Normalized proliferation of MC3T3, MSC, and C4-2B cells in the BME group (n = 3), calculated from fluorescence intensity and expressed as fold-change relative to Day 1. c) A quantitative comparison of the relative proliferation (normalized to Day 1) of C4-2B tumor spheroids between the control group and the BME group over time (Days 1, 7, and 14). d) Schematic of flow cytometry-based sorting and cell cycle analysis of C4-2B cells. e) Cell cycle distribution of C4-2B cells from both groups by flow cytometry. f, g, h) Heatmaps of DEGs in MSC (f), MC3T3 (g), and C4-2B (h) cells. i) Tumors retrieved from mice in control and BME groups on day 21. j, k, l) Quantification of body weight (j), tumor volume (k), and tumor weight (l) in mice from both groups (n = 5). m)In vivo bioluminescent imaging at days 0, 7, 14, and 21. n, o) Histological and molecular analysis of tumors: H&E staining (n) and Ki-67 immunohistochemistry (o). Statistical significance was assessed using one-way ANOVA (n = 3). Significance levels: ns (not significant), *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, respectively.