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–C) Cell adhesion, relative fluorescence intensity of CCR7/CD206 and ROS for Raw264.7 on different Gaussian curvatures. (D) CCR7 & CD206 staining of Raw264.7 cells (blue-nucleus, red-CCR7, green-CD206, scale:100 μm). (E, F) F-actin & DAPI staining and statistical scatterplot of hBMSC adhesion. (G) ALP intensity of hBMSCs on different Gaussian curvatures. (H, I) Cell adhesion and CD31 fluorescence intensity of HUVECs. (J) Chord diagram for comprehensive normalized data of Raw264.7, hBMSCs and HUVECs behavior detection with different Gaussian curvatures on HCGC chips.

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: Fluorescence, Staining

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: Macrophage polarization analysis of Raw264.7 on structures with different gaussian curvature: (A, B) Chord Diagram for qPCR analysis of CCR7, IL6, iNOS-inflammatory and M1 marker genes, and Arg-1, CD206, IL10-M2 related protein genes in different Gaussian curvature groups. (C) Protein content of Arg-1 in different Gaussian curvature groups at 1 and 3 days. (D) Integral plots of the five experimental groups. IL4 group is the positive control for CD206 expression and lipopolysaccharide (LPS) group is the negative control.

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: Marker, Positive Control, Expressing, Negative Control

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

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: Immune-related analysis of macrophages in scaffolds with various Gaussian curvature after muscle bag implantation: (A, B) Immunohistochemical sections and statistical analysis of IL10 positive cells. (C, D) Immunofluorescence staining images and statistical analysis of CD68, CD206 and CCR7 positive cells. (E) The M2-phenotypical genes and inflammatory genes expression of macrophage (n = 3, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001).

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: Immunohistochemical staining, Immunofluorescence, Staining, Expressing

Journal: Materials Today Bio

Article Title: Sequential delivery of sinigrin and dabigatran by an in situ self-stabilizing dynamic hydrogel attenuates intervertebral disc degeneration

doi: 10.1016/j.mtbio.2026.102827

Figure Lengend Snippet: Metabolic regulation of nucleus pulposus vells by SIN and DAB through MAPK and NF-κB pathways. (A) Volcano plot analysis of DE genes (TNF-α vs. TNF-α+SIN). (B) KEGG enrichment analysis of DE genes (TNF-α vs. TNF-α+SIN). (C) GSEA indicated MAPK signalling pathway enrichment. (D) Heatmap of ten selected differentially expressed genes (TNF-α vs. TNF-α+SIN). (E) Western blot analysis of key proteins in the MAPK signalling pathway (p-ERK, p-JNK, p-p38, ERK, JNK, and p38) in the different treatment groups. (F) Quantitative analysis of the Western blot data for MAPK signalling pathway proteins. (G–H) Western blot and quantitative analysis of p-ERK, p-JNK, p-p38, ERK, JNK, and p38 treated with or without TNF-α in the absence or presence of SIN (10 μM) (PCDNA 3.1 vs. TNFR1-OE).(I–L) Integrated analysis of TNF-α vs. TNF-α+DAB DEGs: Volcano plot of DE genes (significance/fold change), KEGG pathway enrichment of DE genes, NF-κB signalling enrichment in GSEA, and heatmap of ten selected differentially expressed genes. (M) Western blot analysis of key proteins in the NF-κB signalling pathway (p-IκBα, p-p65, IκBα, and p65) in the different treatment groups. (N) Quantitative analysis of the Western blot data for the NF-κB signalling pathway proteins. (O–P) Western blot and quantitative analysis of p-p65 and p65 (PCDNA 3.1 vs. TNFR1-OE). (Q) Matched diagram of b and y ions. (R) Western blot analysis of RELA in the different treatment groups. (S) Western blot analysis confirmed the stabilization of p65 by DAB. (T) The affinity between DAB and the p65 target protein was estimated after incubation at different temperatures via Western blot analysis. (n = 3; ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001).

Article Snippet: The membranes were then incubated overnight with anti-GAPDH (1:50 000, AC054, abclone, China), anti-Col2α1(1:500, A19308, abclone, China), anti-Acan (1:1000, A8536, abclone, China), anti-SOX9 (1:1000, A19710, abclone, China), anti-MMP13 (1:1000, A1606, abclone, China), anti-ADAMTS4 (1:500, A2525, abclone, China), anti-iNOS (1:1000, 18985-1-AP, proteintech, China), anti-COX2 (1:1000, 12375-1-AP, proteintech, China), anti-ERK (1:2000, 11257-1-AP, proteintech, China), anti-Phospho-ERK (1:1000, 28733-1-AP, proteintech, China), anti-JNK (1:500, 51153-1-AP, proteintech, China), anti-Phospho-JNK (1:1000, 80024-1-RR, proteintech, China), anti-p38 (1:1000, A4771, abclone, China), anti-Phospho-p38 (1:500, AP1508, abclone, China), anti-p65 (1:5000, A19653, abclone, China), anti-Phospho-p65 (1:500, AP0475, abclone, China), anti-IκBα (1:500, A1187, abclone, China), anti-Phospho-IκBα (1:500, AP0420, abclone, China), anti-AMPK (1:5000, A28094, abclone, China), anti-Phospho-AMPK (1:500, AP0432, abclone, China), The membrane was incubated with HRP-conjugated Goat anti-rabbit IgG (H + L) (1:50 000, AS014, rabbit monoclonal, Abclone, China) and immunoreactive proteins were detected using enhanced chemiluminescence.

Techniques: Western Blot, Incubation

Journal: Bioactive Materials

Article Title: Mossy-textured hydroxyapatite-modified poly (lactic-co-glycolic acid) microspheres promote collagen regeneration via calcium/TGF-β and chemokine signaling pathways in soft tissue augmentation

doi: 10.1016/j.bioactmat.2025.12.028

Figure Lengend Snippet: The regulatory effects of CaHA/PLGA microspheres on macrophages and ADSCs in vitro. (A, B) CLSM images and RFI of CD86 and CD206 expression in RAW264.7 cells co-cultured with microspheres for 2 days (n = 3). (C–F) Relative mRNA expression levels of inflammation-related genes TNF-α, IL-6, TGF-β1, and FGF-2 in RAW264.7 cells (n = 3). (G, H) Sirius red staining images and quantitative analysis (n = 3) of collagen deposition of ADSCs co-cultured with CaHA/PLGA microspheres for 3 and 7 days. (I–K) Relative mRNA expression levels of TGF-β1, FGF-2, and PDGF-A in ADSCs after 3 and 7 days of co-culture with CaHA/PLGA microspheres (n = 3). ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001; ns , not significant.

Article Snippet: CD86 polyclonal antibody (1:200, 13395-1-AP, Proteintech), CD206 (1:200, 18704-1-AP, Proteintech), and TNF-α recombinant antibody (1:200, 80258-6-RR, Proteintech) were used as primary antibodies, incubated overnight at 4 °C.

Techniques: In Vitro, Expressing, Cell Culture, Staining, Co-Culture Assay

Journal: Bioactive Materials

Article Title: Mossy-textured hydroxyapatite-modified poly (lactic-co-glycolic acid) microspheres promote collagen regeneration via calcium/TGF-β and chemokine signaling pathways in soft tissue augmentation

doi: 10.1016/j.bioactmat.2025.12.028

Figure Lengend Snippet: Evaluation of soft tissue filling and inflammatory response in rats. (A, B) Schematic diagram of the soft tissue filling experiment and injection sites, with at least 2 cm spacing between sites. (C) Photographs of subcutaneous soft tissue filling at 2, 4, 8, and 12 weeks. The white circles indicate the soft tissue filling areas. (D) H&E staining of the filled sites. (E, F) Immunofluorescence images and RFI of TNF-α and TGF-β expression at 2 weeks post-filling (n = 6). (G, H) Immunofluorescence images and RFI of CD86 and CD206 expression at 2 weeks post-filling (n = 6). ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001; ns , not significant.

Article Snippet: CD86 polyclonal antibody (1:200, 13395-1-AP, Proteintech), CD206 (1:200, 18704-1-AP, Proteintech), and TNF-α recombinant antibody (1:200, 80258-6-RR, Proteintech) were used as primary antibodies, incubated overnight at 4 °C.

Techniques: Injection, Staining, Immunofluorescence, Expressing