inos  (Bioss)

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: Bacteria-responsive DNAgel system for targeted delivery of photothermally enhanced MXene/MoS 2 in the treatment of pyogenic osteomyelitis

doi: 10.1016/j.bioactmat.2025.10.023

Figure Lengend Snippet: MXMoS 2 DNAgel alleviates inflammation and promotes osteogenesis by modulating macrophage polarization. (A) qRT-PCR analysis of NOS 2 , Il6 , Arg 1 and Il10 levels of RAW264.7 cells after 24h of inflammatory stimulation. (B) ELISA quantification of IL-6 and IL-10 levels of RAW264.7 cells after 24h of inflammatory stimulation. (C – D) Quantitative analysis and representative immunofluorescence staining of iNOS and ARG1. (E) Representative dot plots showing the distribution of iNOS and ARG1 expression in macrophages isolated from POM C57BL/6 mice after DNA hydrogel, MXene/MoS 2 DNA hydrogel, and MXene/MoS 2 DNA hydrogel with NIR irradiation treatment. (F) The stacked bar graphs summarize the proportions of four macrophage subsets (iNOS + ARG1 - , iNOS − ARG1 + , iNOS − ARG1 - , iNOS + ARG1 + ) across groups (n = 3). (G) qRT-PCR analysis of Alp , Runx2 and Sp7 levels of MC3T3-E1 cells after 7 days of osteogenic induction under different treatments. (H) ALP staining of MC3T3-E1 after 7 days. (I) Quantitative analysis of ALP staining. (NC: control group, PC: LPS-treated group) ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001.

Article Snippet: RAW264.7 cells cultured under different stimulation conditions were fixed with 4 % paraformaldehyde (PFA) for 10 min, followed by permeabilization with 0.25 % Triton X-100 in PBS for 10 min. After washing 3 times with PBS (5 min each), cells were blocked with 1 % BSA for 30 min. After washing, cells were incubated with diluted Mouse Arg1 and iNOS antibodies (Proteintech) at room temperature for 1 h. Following additional PBS washes, cells were incubated with secondary antibodies at room temperature for 1 h in the dark.

Techniques: Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Staining, Expressing, Isolation, Irradiation, Control

Journal: Bioactive Materials

Article Title: Bacteria-responsive DNAgel system for targeted delivery of photothermally enhanced MXene/MoS 2 in the treatment of pyogenic osteomyelitis

doi: 10.1016/j.bioactmat.2025.10.023

Figure Lengend Snippet: MXMoS 2 DNAgel alleviates inflammation and promotes osteogenesis by modulating macrophage polarization. (A) qRT-PCR analysis of NOS 2 , Il6 , Arg 1 and Il10 levels of RAW264.7 cells after 24h of inflammatory stimulation. (B) ELISA quantification of IL-6 and IL-10 levels of RAW264.7 cells after 24h of inflammatory stimulation. (C – D) Quantitative analysis and representative immunofluorescence staining of iNOS and ARG1. (E) Representative dot plots showing the distribution of iNOS and ARG1 expression in macrophages isolated from POM C57BL/6 mice after DNA hydrogel, MXene/MoS 2 DNA hydrogel, and MXene/MoS 2 DNA hydrogel with NIR irradiation treatment. (F) The stacked bar graphs summarize the proportions of four macrophage subsets (iNOS + ARG1 - , iNOS − ARG1 + , iNOS − ARG1 - , iNOS + ARG1 + ) across groups (n = 3). (G) qRT-PCR analysis of Alp , Runx2 and Sp7 levels of MC3T3-E1 cells after 7 days of osteogenic induction under different treatments. (H) ALP staining of MC3T3-E1 after 7 days. (I) Quantitative analysis of ALP staining. (NC: control group, PC: LPS-treated group) ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001.

Article Snippet: Primary antibodies for immunocytochemistry and immunofluorescence staining—including ARG1 and iNOS, TRAP, β-Catenin, OCN and WNT4—were obtained from Proteintech (China).

Techniques: Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Staining, Expressing, Isolation, Irradiation, Control

Journal: The Journal of Experimental Medicine

Article Title: Tet2 deficiency–induced expansion of monocyte-derived macrophages promotes liver fibrosis

doi: 10.1084/jem.20251114

Figure Lengend Snippet: Aging-related Tet2 ΔMye -driven clonal hematopoiesis exacerbates liver fibrosis, and IL-1β–NLRP3 pathway is not involved in the liver fibrosis in Mye-KO-CCl 4 mice. (A) Chart for the construction of an aging-related Tet2 ΔMye -induced clonal hematopoiesis mouse model with liver fibrosis. BDL was performed to construct a liver fibrosis model in young-BMT and old-BMT mice 6 wk after BMT. (B) Frequency of MDMs in livers of young-BMT and old-BMT mice after BDL for 4 wk ( n = 4 for each group). (C) The serum levels of CCL2, CCL8, and IL-6 in young-BMT and old-BMT mice after BDL for 4 wk ( n = 4 for each group). (D and E) Picrosirius red staining (D) (scale bar, 125 μm) and quantitative analysis of collagen deposition (E) in livers from young-BMT and old-BMT mice following BDL, treated with Bindarit plus anti–IL-6 Abs ( n = 4 for each group). (F and G) Changes of serum Col IV (F) and HA (G) levels in young-BMT and old-BMT mice treated with Bindarit, anti–IL-6 Abs, or Bindarit plus anti–IL-6 Abs ( n = 4 for each group). (H) IF (scale bar, 50 μm) and statistical analysis of NLRP3 in the liver of Tet2 WT -CCl 4 and Tet2 ΔMye -CCl 4 mice ( n = 5 for each group). Red: anti-NLRP3; blue: DAPI. (I) IHC staining (scale bar, 50 μm) and statistical analysis of NLRP3 in the liver of Tet2 WT -CCl 4 and Tet2 ΔMye -CCl 4 mice ( n = 5 for each group). (J) Inhibition of IL-1β in the serum of oil- or CCl 4 -treated Tet2 WT and Tet2 ΔMye mice ( n = 5 for each group). (K and L) Change of Ccl2 (K) and Ccl8 (L) in the serum of oil- or CCl 4 -treated Tet2 WT and Tet2 ΔMye mice after anti–IL-1β Ab treatment ( n = 4–5 for each group). (M and N) Effect of anti–IL-1β Ab treatment on the mRNA levels of Acta2 and Col1a1 ( n = 5 for each group). (O) Picrosirius red staining of collagen deposition in the liver tissue of Tet2 WT -CCl 4 and Tet2 ΔMye -CCl 4 mice after treatment of anti–IL-1β Ab ( n = 5 for each group). Data are representative of at least two independent experiments with similar results (B, C, and E–O). All data are shown as mean ± SD and were analyzed by two-tailed, paired Student’s t test (B, C, and H, and I) or two-way ANOVA with Sidak’s multiple comparison test (E–G and J–N). ***P < 0.001; **P < 0.01; *P < 0.05; P > 0.05 not significant (ns).

Article Snippet: Primary Abs against CD45.2 (60287-1-Ig; Proteintech), F4/80 (ab300421; Abcam), Ly6c (65296-1-Ig; Proteintech), CD68 (ab53444; Abcam), CD206 (ab300621; Abcam), Inos (22226-1-AP; Proteintech), and NLRP3 (22226-1-AP; Proteintech) were applied and incubated overnight at 4°C.

Techniques: Construct, Staining, Immunohistochemistry, Inhibition, Two Tailed Test, Comparison