Fig 1: IL1ß-induced HyC enhances in vitro human osteoclast differentiation. (A–H) Brightfield images and TRAP staining in in vitro osteoclast cultures in presence of control (“C”) and IL1ß-induced (“IL1ß”) HyC matrices with (+) or without (-) RANKL. Scale bar, 500 µm. (I) TRAP quantification in osteoclast cultures in presence of control and IL1ß-induced HyC matrices with/without RANKL. (J–K) Gene expression of (J) MMP9 and (K) CTSK in osteoclast cultures in presence of control and IL1ß-induced HyC matrices with/without RANKL. (I–K) Data are plotted as means ± SD; n = 6. *P < 0.05, **P < 0.01, ****P < 0.0001. One-way ANOVA with Tukey's multiple comparison tests.
Fig 2: IL1ß-induced HyC promotes in vivo vascular recruitment, osteoclast activity and M2 macrophage polarization. (A–B) Immunofluorescent staining for CD31 (red) in 10 µm-sections of tissues derived from control and IL1ß-induced matrices after 1 week of in vivo remodeling. MSOD-B-derived cells are labelled in green (GFP expression). Nuclei are labelled with DAPI (blue). The dashed lines delimitate the human cartilaginous tissue with MSOD-B cells from the surrounding mouse connective tissue. Scale bar, 100 µm. (C) Quantification of CD31+ area in remodeled tissues after 1 week in vivo. (D–E) Immunostaining for Osterix in 10 µm-sections of tissues derived from control and IL1ß-induced matrices after 1 week of in vivo remodeling. Scale bar, 100 µm. (F) Quantification of Osterix + area in remodeled tissues after 1 week in vivo. (G–H) Tartrate-resistant acid phosphatase (TRAP) staining in tissues derived from control and IL1ß-induced matrices after 1 week of in vivo remodeling. Scale bar, 100 µm. (I) Quantification of TRAP + area in remodeled tissues after 1 week in vivo. (J–N) Percentage of (J) CD11b+Ly6G- monocytes, (K) CD11b+Ly6G+ granulocytes, (L) CD11b+F4/80+ macrophages, (M) CD11b+F4/80+CD206+ M2 macrophages and (N) CD11blowcKit + CD115+ osteoclast precursors. (C, F, I, J-N) Data are plotted as means ± SD; n = 5. *P < 0.05, **P < 0.01. Unpaired two-tailed t-test.
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