Journal: Bioactive Materials
Article Title: Geometry-driven immunomodulation in 3D-printed bioceramics: Negative curvature promotes macrophage M2 polarization via Ras-MAPK/HIF-1α signaling for vascularized osteogenesis
doi: 10.1016/j.bioactmat.2026.01.001
Figure Lengend Snippet: (A) Immunofluorescence images of β-Tubulin /CD206 and β-Tubulin /HIF-1α of macrophages in different Gaussian curvature groups before and after treatment with Adezmapimod . (B, C) Protein quantification and Western blotting image of ERK1/2, p-ERK1/2, β-tubulin, HIF-1α and CD206 proteins in all groups before and after treatment with Adezmapimod and Paclitaxel. (D) Mechanism summary of Gaussian curvature-driven macrophage polarization.
Article Snippet: The reagents used in the experiment included: H-DMEM(11965118, Gibco, USA.), α-DMEM medium(12571063, Gibco, USA.), TritonX-100(ST1723, Beyotime, China), 4 % paraformaldehyde (BL539A, Biosharp, China),FBS(A5256701, Gibco, USA.),ECM medium (Science Cell, USA.),and DAPI staining solution (C1006, Beyotime, China),BCIP/NBT(C3206, Beyotime, China), reactive oxygen species kit (S0033S, Beyotime, China), BSA (B2064, ≥98 %, Sigma-Aldrich, USA.),CD31 antibody (ab28364, Abcam, USA.), secondary anti-IGg (ab175773, Alexa Fluor® 680, Abcam, USA.), Phalloidin-iFluor 488(ab176753, Abcam, USA.), CCR7(AF5293, Bioss, China), CD206 (bsm-60761R, Bioss, China), iNOS (bs-22924R, Bioss, China), RIPA (P0013, Beyotime, China), p-ERK1/2 (AF3687, Affinity, USA.) and ERK1/2 (#AF0155, Affinity, USA), luminol detection reagent (sc-2048, Santa Cruz, USA.), GAPDH (Cat#KC-5G5, Kangchen Biotechnology, China), Trizol(15596026CN, Invitrogen, USA.), DEPC(R0601, Thermo Scientific, USA.), TBST(R017R.0000, Thermo Scientific, USA.), HIF-1a(GTX127309, GeneTex, USA.), β-Tubulin(10094-1-AP, Proteintech, UK) Adezmapimod (SB 203580, MCE, USA.) medium and Paclitaxel (99.88 %, HY-B0015R, MCE), ELISA Arg-1(E-EL-M3092, ELabSci@, China), TNF-α (E-EL-M3063, ELabSci@, China), OPN(22952-1-AP, Proteintech, UK.), F4/80 (GB11027-100, Servicebio, China) Alkaline phosphatase activity kit (P0321S, Beyotime, China), Matrigel (CLS356234, Corning, USA), Microfill MV120 (Flow tech, USA), EDTA(17892, Thermo Scientific, USA.) xylene(X112051, AR,99 %, Aladdin, China), ethanol (107-21-1, AR,99 %, Aladdin, China).
Techniques: Immunofluorescence, Western Blot