rank l  (R&D Systems)

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: Journal of Human Immunity

Article Title: Human ISG15 deficiency unveils impaired healing of ulcerations via type I interferon–mediated fibrosis

doi: 10.70962/jhi.20250011

Figure Lengend Snippet: Myofibroblast activation in ISG15-deficient cells. (A) Spatial expression and intensity levels of two myofibroblast markers ( ACTA2 and COL5A1 ). (B) Relative expression of various myofibroblast markers for clusters 1 and 5 separated by sample biopsy. (C) Percentage of total spots that express TGFβ in the entire biopsy, within CD68 + spots only, and within myofibroblast cluster 5. (D) Significance of pathway enrichment for clusters 0, 1, 5, 3, 7, and 9 generated by MSigDB Hallmark 2020 and BioPlanet 2019 analysis from Enrichr. Adjusted P values are shown. (E) Relative RNA levels of myofibroblast markers ( PDGFA , ITGB6 , FGF2 ) following a 72-h treatment with 1,000 IU/ml IFNα2b, 10 ng/ml TGFβ, or both of WT and ISG15 KO lung epithelial cells (A549). (F) Percentage of cells undergoing EMT upon TGFβ/IFNα stimulation. (G) Relative RNA levels of myofibroblast markers ( PDGFA , ITGB6 , FGF2 ) following a 5-day treatment with 1,000 IU/ml IFNα2b, 10 ng/ml TGFβ, or both of WT and ISG15 KO fibroblasts. (H) ACTA2 (myofibroblasts) IHC for lesion 1 and lesion 3. P values were calculated with two-tailed t test. *P < 0.05.

Article Snippet: The following day, cells were treated with IFN-α2b (100 or 1,000 IU ml −1 , Merck IntronA), TGFβ (10 ng ml −1 , 75362; Cell Signaling), or both IFNα and TGFβ together.

Techniques: Activation Assay, Expressing, Generated, Two Tailed Test

Journal: Bioactive Materials

Article Title: Carrier free oral Co-delivery of atorvastatin via baicalein-copper-network for atherosclerosis therapy through senescence reversal and multi-mechanistic synergy

doi: 10.1016/j.bioactmat.2025.12.036

Figure Lengend Snippet: Reactive oxygen species scavenging capacity of CMA. (A) H 2 O 2 (100 μM) scavenging capacity CMA at 2 h. (B) O 2 − scavenging capacity of CMA at 40 min. (C) •OH scavenging capacity of CMA at 1 min. (D) DPPH (100 μg mL -1 ) scavenging capacity of CMA at 1 h. (E) Fluorescence images of RAW264.7 cells treated with CMA and LPS + IFN-γ for 24 h. Intracellular ROS were stained with DCFH-DA (Green), and nuclei were stained with DAPI (blue). Scale bar = 50 μm. (F) Quantitative analysis of mean DCFH-DA fluorescence intensity. Flow cytometric analysis of ROS expression in: (G) RAW264.7: CMA + LPS; (H) RAW264.7: CMA; (I) SMC: CMA; (J) HUVEC: CMA; (K) RAW264.7: CMA + BAPTA; (L) SMC: CMA + BAPTA; (M) HUVEC: CMA + BAPTA. All treatments were conducted for 24 h. Data are presented as mean ± SD (n = 3–6; ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001).

Article Snippet: Baicalein (BAI), copper chloride (CuCl 2 ·2H 2 O), and Atorvastatin (ATV) were purchased from Macklin Inc. Lipopolysaccharides (LPS), recombinant mouse interferon γ (IFN-γ), oxidized low-density lipoprotein (oxLDL), dihydroethidium (DHE), DiI-oxidized low-density lipoprotein (DiI-oxLDL), hematoxylin-eosin (H & E) stain kit, modified Masson's trichrome stain kit, and modified Oil Red O stain kit were obtained from Beijing Solarbio Science & Technology Co., Ltd. Cy5-baicalein was purchased from Xi'an Qiyue Biology.

Techniques: Fluorescence, Staining, Expressing

Journal: Bioactive Materials

Article Title: Carrier free oral Co-delivery of atorvastatin via baicalein-copper-network for atherosclerosis therapy through senescence reversal and multi-mechanistic synergy

doi: 10.1016/j.bioactmat.2025.12.036

Figure Lengend Snippet: Macrophage reprogramming ability of CMA. (A) Representative optical images of RAW264.7. Scale bar = 50 μm. (B) Confocal laser scanning microscopy images of RAW264.7 cells stained with CD206 antibody, with nuclei counterstained with DAPI. Scale bar = 50 μm. (C) Quantitative analysis of mean CD206 fluorescence intensity. (D) Flow cytometric analysis of CD206 expression in RAW264.7 treated with BAI, Cu-PBS, Cu-MON, ATV, and CMA for 24 h. (E) Relative expression levels of Arg-1, VEGF, TNF-α, and IL-1β in cell supernatants. (F) Flow cytometry scatter plots of iNOS and CD206 expression in RAW264.7 pretreated with LPS + IFN-γ for 24 h followed by treatment with BAI, Cu-PBS, Cu-MON, ATV, and CMA for 24 h. (G–I) Flow cytometric analysis of M1/M2 expression (MFI ratio), iNOs expression, CD206 expression in RAW264.7. (J) Relative expression levels of TGF-β, Arg-1, VEGF, TNF-α, and IL-1β in supernatants from cells treated as in F. Data represent mean ± SD (n = 3–6 independent experiments). Statistical significance: ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001 versus Control; ns = not significant.

Article Snippet: Baicalein (BAI), copper chloride (CuCl 2 ·2H 2 O), and Atorvastatin (ATV) were purchased from Macklin Inc. Lipopolysaccharides (LPS), recombinant mouse interferon γ (IFN-γ), oxidized low-density lipoprotein (oxLDL), dihydroethidium (DHE), DiI-oxidized low-density lipoprotein (DiI-oxLDL), hematoxylin-eosin (H & E) stain kit, modified Masson's trichrome stain kit, and modified Oil Red O stain kit were obtained from Beijing Solarbio Science & Technology Co., Ltd. Cy5-baicalein was purchased from Xi'an Qiyue Biology.

Techniques: Confocal Laser Scanning Microscopy, Staining, Fluorescence, Expressing, Flow Cytometry, Control