Journal: Bioactive Materials
Article Title: Immune regulative GelMA&Zn 2+ /Ce 3+ -whitlockite scaffolds with continuous ions release for bone regeneration
doi: 10.1016/j.bioactmat.2025.11.009
Figure Lengend Snippet: Evaluation of immune inflammation (RAW264.7) in the scaffolds. A, B) Immunofluorescent staining results and quantitative fluorescence intensity of CD206 in RAW264.7 cells co-cultured with different sample extracts, with CD206 (M2 marker) shown in red (scale bar = 50 μm). C, D) Immunofluorescent staining results and quantitative fluorescence intensity of CD86 in RAW264.7 cells co-cultured with different sample extracts, with CD86 (M1 marker) shown in green (scale bar = 50 μm). E, F) Immunofluorescent staining results and quantitative fluorescence intensity of CD163 in RAW264.7 cells co-cultured with different sample extracts, with CD163 (M2 marker) shown in green (scale bar = 50 μm). G) Flow cytometry analysis of CD86 and CD11b positive cells in inflammatory RAW264.7 cells. H) Fluorescence enzyme labeling for the quantitative analysis of ROS content in macrophages. I) Cell viability of RAW264.7 cells co-cultured with different sample extracts for 1, 3, and 5 days. J) NO content in RAW264.7 cells co-cultured with different sample extracts. K) DCFH-DA fluorescent probe detection to evaluate ROS content in macrophages after stimulation with different sample extracts (scale bar = 100 μm). P values were determined using one-way ANOVA (∗∗∗∗P < 0.0001, ∗∗∗P < 0.001, ∗∗P < 0.01, ∗P < 0.05, N.S., not significant).
Article Snippet: Following stimulation, the cells were blocked with 5 % bovine serum albumin (BSA) for 2 h at room temperature, washed three times with PBS, and incubated overnight at 4 °C with a CD86 polyclonal antibody (1:600, 13395-1-AP, Proteintech, USA) and CD163 polyclonal antibody (1:600, 16646-1-AP, Proteintech, USA).
Techniques: Staining, Fluorescence, Cell Culture, Marker, Flow Cytometry, Labeling
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