raw264 7  (ATCC)

Journal: Bioactive Materials

Article Title: Bioengineered titanium implants functionalized with aptamer-valproic acid conjugates orchestrate macrophage programming and mesenchymal stem cell homing for improved osseointegration

doi: 10.1016/j.bioactmat.2026.05.055

Figure Lengend Snippet: A) Live/Dead staining of RAW 264.7 macrophages cultured on different material surfaces for 72 h, where dead cells were stained red and live cells were stained green. B) Cytoskeleton staining morphology of RAW 264.7 grown on different material surfaces for 72 h. C, D, F) Fluorescence images and flow cytometry analysis of intracellular ROS in RAW 264.7 cells with DCFH-DA probe. E) Proliferation of RAW 264.7 macrophages on different material surfaces assessed by CCK-8 assay. n = 3. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Created in BioRender.

Article Snippet: The murine macrophage cell line RAW 264.7 (Procell Life Science, China) was cultured in high-glucose DMEM (Gibco, USA) containing 10% FBS and employed for immunomodulation studies.

Techniques: Staining, Cell Culture, Fluorescence, Flow Cytometry, CCK-8 Assay

Journal: Bioactive Materials

Article Title: Bioengineered titanium implants functionalized with aptamer-valproic acid conjugates orchestrate macrophage programming and mesenchymal stem cell homing for improved osseointegration

doi: 10.1016/j.bioactmat.2026.05.055

Figure Lengend Snippet: A, F) IF staining of iNOS and CD206 in RAW 264.7 macrophages. B-E) Secretion of inflammation-related proteins in RAW 264.7 macrophages. G-J) Relative mRNA expression of inflammation-related genes in RAW 264.7 macrophages. n = 3. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: The murine macrophage cell line RAW 264.7 (Procell Life Science, China) was cultured in high-glucose DMEM (Gibco, USA) containing 10% FBS and employed for immunomodulation studies.

Techniques: Staining, Expressing

Journal: Bioactive Materials

Article Title: Chronic inflammation-responsive hydrogel restores myeloid-T cell crosstalk to reinvigorate antitumor immunity against metastatic colorectal cancer

doi: 10.1016/j.bioactmat.2026.03.012

Figure Lengend Snippet: PGE2 blockade modulates immune cell phenotypes in antitumor resp onses. (A) Inflammatory gene expression across cancer types (GEPIA2 database). (B) Gene expression of Il1b , Cxcl8 , and Lif in colon adenocarcinoma (COAD) tumor tissue and normal tissue (GEPIA2 database). (C and D) Correlation between Ptgs2 and inflammatory genes in various cancers (C) and COAD (D) (TIMER 2.0). (E) Schematic of immune cells co-incubated with CXB treated tumor conditional medium (TCM) (Source material from BioRender). (F and G) Cell viability (F) and Cell cycle arrest (G) detection of CT26 tumor cells treated with gradient concentrations of CXB; n = 3. (H) PGE2 concentration in CT26 cell supernatants; n = 3. (I) The proportion of CD103 + DC within BMDCs after CXB treatments in vitro ; n = 3. (J and K) Maturation (J) and Antigen processing capability (K) on BMDCs; n = 3. (L – N) Flow charts of CD86 or CD206 expression on Raw 264.7 cells (L). Quantification of CD86 (M) and CD206 (N) expression on Raw 264.7 cells; n = 3. (O and P) Flow charts (O) and Quantification (P) of CD69 and CD137 expression on splenic T cells exposed to CXB-pretreated TCM; n = 3. (Q) IFN-γ secretion by T cells co-cultured with CXB-pretreated TCM; n = 3. Data are presented as mean ± SD, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. Significance was calculated using One-way ANOVA.

Article Snippet: CT26 cells and Raw 264.7 were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Gene Expression, Incubation, Concentration Assay, In Vitro, Expressing, Cell Culture

Journal: Bioactive Materials

Article Title: Chronic inflammation-responsive hydrogel restores myeloid-T cell crosstalk to reinvigorate antitumor immunity against metastatic colorectal cancer

doi: 10.1016/j.bioactmat.2026.03.012

Figure Lengend Snippet: TRANS modulates macrophage polarization and promotes antigen present ation. (A) Uniform manifold approximation and projection (UMAP) visualization of myeloid cells in the TME following PBS or TRANS treatment. (B) UMAP clusters of myeloid cells. (C) Cell-type-specific marker genes expression of different clusters. (D) Proportion distribution of myeloid cell clusters. (E) KEGG enrichment in M1 macrophages (TRANS vs PBS). (F) GSEA of NOD-like receptor and JAK-STAT signaling pathways in M1. (G) KEGG enrichment in M2 macrophages (TRANS vs PBS). (H) Glycolysis/Gluconeogenesis and antigen processing/presentation in M2. (I) Scheme of M0 macrophages treated with different TCM. (J) M1 and M2 polarization from M0 macrophages under various treatments; n = 3. (K) STAT1 expression in Raw 264.7 cells. (L) Scheme of M2 macrophages treated with different TCM. (M) Repolarization of M2 macrophages under different treatments; n = 3. (N) H-2K b /SIINFEKL + macrophages in vitro ; n = 3. (O and P) Relative expression of antitumor genes (O) and inflammatory genes (P) in macrophages under different treatments in vitro ; n = 3. (Q) CD69 expression on T cells after co-incubating TCM-treated macrophages and anti-CD3/CD28-stimulated splenic T cells; n = 3. Data are presented as mean ± SD, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. Significance of multiple-group comparisons was calculated using One-way ANOVA. Significance of two-group comparison was calculated using Student's t -test.

Article Snippet: CT26 cells and Raw 264.7 were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Marker, Expressing, Protein-Protein interactions, In Vitro, Comparison

Journal: Bioactive Materials

Article Title: Chiral Fe 3 O 4 /GelMA hydrogels regulate the osteoimmune microenvironment via Itgb3-mediated macrophage polarization to combat peri-implantitis

doi: 10.1016/j.bioactmat.2026.03.055

Figure Lengend Snippet: Single-cell and immune-profiling analysis reveals an imbalanced osteo-immune microenvironment in peri-implantitis. (A) Immune infiltration scores of peri-implant tissues. (B) UMAP visualization of peri-implant tissues, colored by cell-type ontology, showing the distribution of major cell populations. (C, D) Expression patterns of key markers Mrc1 and Tnf projected onto the UMAP space. The color gradient from yellow to blue indicates expression levels from low to high. (E) Dot plot showing the expression of M1-associated ( Tnf , Il6 , Nos2 ) and M2-associated ( Mrc1 , Arg1 , Il10 ) macrophage signature genes. The dot size represents the percentage of cells expressing the gene, and color intensity indicates the average expression level. (F, G) Violin plots illustrating the expression distribution of M1/M2 signature genes across different cell types. (H) Osteogenic genes ( Alpl , Col1a1 ) are specifically and highly expressed in osteoblast clusters. Abbreviation: UMAP, Uniform Manifold Approximation and Projection.

Article Snippet: Macrophages (RAW 264.7, ATCC) were cultured in Dulbecco's Modified Eagle Medium (DMEM; Gibco, USA) supplemented with 10% fetal bovine serum (FBS; HyClone, USA) and 1% penicillin-streptomycin (Gibco, USA) at 5% CO 2 and 37 °C.

Techniques: Single Cell, Expressing

Journal: Bioactive Materials

Article Title: Chiral Fe 3 O 4 /GelMA hydrogels regulate the osteoimmune microenvironment via Itgb3-mediated macrophage polarization to combat peri-implantitis

doi: 10.1016/j.bioactmat.2026.03.055

Figure Lengend Snippet: In vitro experiments validate the mechanism by which chiral hydrogels modulate the bone immune microenvironment. (A) GO and (B) KEGG enrichment analysis of DEGs with elevated D-FG relative to L-FG. (C) Protein expression levels of p-AKT and AKT after 48-h co-culture with different hydrogels. (D, E) Intracellular ROS fluorescence levels and quantitative analysis after SC79-mediated PI3K/Akt activation. (F, G) ROS levels and quantification in RAW264.7 cells after LPS stimulation and 24 h co-culture with different hydrogels. (H, I) ROS levels and quantification in RAW264.7 cells after 72 h co-culture with different hydrogels. (J, K) Representative fluorescence images and quantitative analysis of Fe 2+ release and ROS kinetics in chiral hydrogel-treated macrophages. Scale bar = 50 μm. (n = 3, compared with the control group, ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 vs. GelMA group. # P < 0.05, ## P < 0.01, ### P < 0.001). (L) Schematic representation of the bone immunomodulatory mechanism of chiral hydrogels. Abbreviation: KEGG, Kyoto Encyclopedia of Genes and Genomes; p-AKT, phosphorylated AKT; AKT, protein kinase B.

Article Snippet: Macrophages (RAW 264.7, ATCC) were cultured in Dulbecco's Modified Eagle Medium (DMEM; Gibco, USA) supplemented with 10% fetal bovine serum (FBS; HyClone, USA) and 1% penicillin-streptomycin (Gibco, USA) at 5% CO 2 and 37 °C.

Techniques: In Vitro, Expressing, Co-Culture Assay, Fluorescence, Activation Assay, Control

Journal: Bioactive Materials

Article Title: Near infrared enhanced palladium loaded siraitia grosvenorii carbon dots amplify mitophagy for acute lung injury immunotherapy

doi: 10.1016/j.bioactmat.2026.02.040

Figure Lengend Snippet: Schematic illustration of the preparation of CPs@SS31, and the corresponding in vivo therapy of acute lung injury via NIR enhanced ROS scavenging, inflammation inhibition, macrophage M2 polarization, and T cells immunoactivation, as well as specifically targeting mitochondria, activating mitochondrial function, and inducing mitophagy to reprogram lung redox homeostasis, and promote tissue repair.

Article Snippet: Cell viability testing : Mouse monocyte macrophages (RAW264.7, ATCC, USA) were cultured in DMEM containing 10% FBS and 1% penicillin-streptomycin (Solarbio, China).

Techniques: In Vivo, Inhibition

Journal: Bioactive Materials

Article Title: Smart microenvironment-adaptive nanocatalytic hydrogel for sequential antibacterial, anti-inflammatory, and regenerative therapy of biofilm-infected wounds

doi: 10.1016/j.bioactmat.2026.02.043

Figure Lengend Snippet: Characterization, and Cytocompatibility Validation of HCOC. (A) Schematic illustration of the development of HCOC. (B) FTIR spectrum of OSA, CMCS and OC hydrogel. (C) Time-dependent evolution of gelation of OC and HCOC. (D) SEM images of HCOC and EDS mapping images of C, N, O and Cu for HCOC. (E) FTIR spectra of HC, OC and HCOC. (F) Dynamic frequency sweep measurements of OC and HCOC. (G) Frequency-dependent viscoelastic behavior of OC and HCOC. (H) Alternating strain sweep with alternating strains of 1% and 1000% at 100s intervals and (I) Self-healing behavior of HCOC. (J) Live/dead staining showing the metabolic activity of L929 and RAW 264.7 cells after treatment with HCOC for 48 h. Rates of proliferation of (K) L929 cells and (L) RAW 264.7 cells after treatment with PBS or HCOC. (∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001).

Article Snippet: L929 and RAW 264.7 cells were purchased from the American-Type Culture Collection (ATCC).

Techniques: Biomarker Discovery, Staining, Activity Assay

Journal: Bioactive Materials

Article Title: Smart microenvironment-adaptive nanocatalytic hydrogel for sequential antibacterial, anti-inflammatory, and regenerative therapy of biofilm-infected wounds

doi: 10.1016/j.bioactmat.2026.02.043

Figure Lengend Snippet: Macrophage immunoregulation, cell proliferation and angiogenesis in vitro triggered by HCOC. (A) Localization of CD86 (orange, M1 marker) and CD206 (green, M2 marker) in RAW 264.7 macrophages after stimulation (24 h) with different test materials (PBS, 40 ng/mL IL-4, 2 mg/mL Cu 5.4 O, 2 mg/mL HAs, 2 mg/mL HC, 2 mg/mL HCOC). (B) Flow cytometric analysis quantifying CD86 + and CD206 + cell populations after 24 h inflammatory stimulation and different concentrations of HCOC. (C) Western blot analysis of key signaling proteins and phenotypic markers in LPS-stimulated RAW264.7 macrophages treated with HCOC (0.5 or 1.0 mg/mL) or IL-4 (20 ng/mL). Protein levels were normalized to GAPDH or the corresponding total protein. (D–G) mRNA expression of (D) TNF-α, (E) Arg1, (F) Axin2, and (G) IL-10 measured by real-time RT-PCR. Data are expressed relative to the LPS-only control group and presented as mean ± SEM of three independent experiments. (H) Following H 2 O 2 pretreatment, L929 cell migration and (I) HUVEC tube formation ( in vitro angiogenesis) were assessed after treatment with varying HCOC concentrations over a 24-h period. Quantitative analysis of (J) L929 cell migration rate and the formed (K tube length and (L) branch points. (∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001).

Article Snippet: L929 and RAW 264.7 cells were purchased from the American-Type Culture Collection (ATCC).

Techniques: In Vitro, Marker, Western Blot, Expressing, Quantitative RT-PCR, Control, Migration