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: 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: 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: A foam cell-targeted lipophagy restoration strategy stabilizes vulnerable atherosclerotic plaques

doi: 10.1016/j.bioactmat.2026.02.041

Figure Lengend Snippet: In vitro evaluation of foam cell lipid accumulation and lipophagy activation following OPN-HMCN@MLT treatment. ( A - C ) ORO and BODIPY staining images and corresponding quantification of ORO and BODIPY positive areas of RAW264.7 cells under different stimulations (n = 5, scale bar for ORO: 100 μm, scale bar for BODIPY: 20 μm). ( D ) Bio-TEM images of RAW264.7 cells post various treatments (n = 5, scale bars 1.0 μm). Green arrows indicate nanoparticles. ( E , F ) Morphometric analysis determined the mean number and area (μm 2 ) of LDs per cell section. ( G ) Confocal images depicting lipophagy flux in foam cells following different treatments (n = 5 biological replicates, scale bars: 10 μm). ( H - J ) The quantities of acidified autophagosomes (GFP-RFP+), neutral autophagosomes (GFP + RFP+), and LDs labeled with BODIPY were measured per cell for each condition. (K to N) Representative Western blot images and quantitative analysis of LC3, LAMP1, and P62 expression in foam cells. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, and ∗∗∗∗ P < 0.0001.

Article Snippet: RAW264.7 mouse macrophage cells (ATCC® TIB-71; RRID: CVCL_0493) and MCAECs (Procell, CP-M081) were cultured in Dulbecco's Modified Eagle Medium (DMEM) with 10% fetal bovine serum and 1% penicillin-streptomycin at 37 °C in a 5% CO 2 atmosphere.

Techniques: In Vitro, Activation Assay, Staining, Labeling, Western Blot, Expressing

Journal: Bioactive Materials

Article Title: A foam cell-targeted lipophagy restoration strategy stabilizes vulnerable atherosclerotic plaques

doi: 10.1016/j.bioactmat.2026.02.041

Figure Lengend Snippet: In vitro examination of LD degradation in foam cells through fatty acid oxidation and cholesterol efflux. (A ) Schematic diagram of the LD degradation mechanism. ( B , C ) Confocal images and quantitative analysis of LDs colocalization with fatty acids in RAW264.7 cells following different treatments (n = 5, scale bars: 5 μm). ( D , E ) Confocal images of the colocalization of mitochondria with fatty acids and quantified data of fatty acids in RAW264.7 cells under different stimulations (n = 5, scale bars: 5 μm). ( F , G ) Confocal images illustrating mitochondrial colocalization with ATP and corresponding quantification of ATP levels in RAW264.7 cells post various treatments (n = 5, scale bars: 20 μm). ( H ) Diagram illustrating the incorporation of [U- 13 C] palmitic acid into the TCA cycle and the labeling pattern of derived metabolites (n = 3). ( I ) A PCA plot illustrates the cluster separation between the two groups (n = 3). ( J ) Heatmap showing differences in metabolites between the two groups (n = 3). ( K ) Normalized total labeling of each metabolite to [U- 13 C] palmitic acid (n = 3). ( L ) Proportion of (m + 2)-labeled TCA cycle metabolites derived from [U- 13 C] palmitic acid (n = 3). ( M - R ) The study quantified NBD-cholesterol accumulation ( M , P ) and cholesterol efflux facilitated by HDL ( N , O ) and apoA-I ( Q , R ) using confocal imaging across (n = 5, Scale bar = 50 μm). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, and ∗∗∗∗ P < 0.0001.

Article Snippet: RAW264.7 mouse macrophage cells (ATCC® TIB-71; RRID: CVCL_0493) and MCAECs (Procell, CP-M081) were cultured in Dulbecco's Modified Eagle Medium (DMEM) with 10% fetal bovine serum and 1% penicillin-streptomycin at 37 °C in a 5% CO 2 atmosphere.

Techniques: In Vitro, Labeling, Derivative Assay, Imaging

Journal: Bioactive Materials

Article Title: Harnessing the gut–immune–joint axis: Oral microalgae-based thermoresponsive microspheres enhance intra-articular therapy for rheumatoid arthritis

doi: 10.1016/j.bioactmat.2026.01.037

Figure Lengend Snippet: Evaluation of the protective effects of CG@GelMA on FSN-induced intestinal epithelial barrier disruption in vitro . (A) Immunofluorescence staining of Claudin-1 in Caco-2 monolayers under different treatments. Scale bar: 100 μm. (B) Relative fluorescence intensity of Claudin-1. (C) Immunofluorescence staining of Occludin in Caco-2 monolayers. Scale bar: 100 μm. (D) Relative fluorescence intensity of Occludin. (E) Immunofluorescence staining of ZO-1 in Caco-2 monolayers. Scale bar: 100 μm. (F) Relative fluorescence intensity of ZO-1. (G) Schematic diagram of the Caco-2/RAW 264.7 Transwell co-culture system. (H) Relative TEER of Caco-2 cell monolayers after different treatments. (I) Relative fluorescence intensity of FD4 across Caco-2 monolayers under different treatments. Data are presented as means ± SD. Statistical significance: ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001.

Article Snippet: RAW 264.7 macrophages (Procell, Wuhan, China), Caco-2 intestinal epithelial cells (Pricella, Wuhan, China), and IEC-6 small intestinal epithelial cells (ATCC, USA) were maintained in high-glucose DMEM supplemented with 10 % fetal bovine serum (AiTing, Hangzhou, China) and 1 % penicillin-streptomycin (Gibco, USA), with the medium for IEC-6 cells additionally containing 0.1 U/mL human insulin.

Techniques: Disruption, In Vitro, Immunofluorescence, Staining, Fluorescence, Co-Culture Assay