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CBNK-EVs rapidly impair ZIKV infectivity and protect host cells. (a) Multiplex immunohistochemical analysis of ZIKV infection in neural progenitor cells. Co-staining for Nestin (neural progenitor cells, green) and ZIKV E protein (red) was performed, and double-positive cells were quantified across six random fields, mean ± SD, ∗P < 0.05. Scale bar, 20 μm. (b) KEGG pathway enrichment bubble plot from <t>RNA-seq</t> <t>of</t> <t>HTR-8/Svneo</t> cells treated with CBNK-EVs (3 × 10 11 particles/mL, 10 μl) and ZIKV (MOI = 1) for 1 h. (c) GSEA analysis of transcriptomic data from HTR-8/Svneo cells. (d) Western blot analysis of ZIKV E and ZIKV NS5 protein levels after co-incubation of ZIKV particles with CBNK-EVs for the indicated time periods (0–30 min) prior to infection. (e) The abundance of NK cell-associated proteins in CBNK-EVs. ∗P < 0.05 (one-way ANOVA).
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CBNK-EVs rapidly impair ZIKV infectivity and protect host cells. (a) Multiplex immunohistochemical analysis of ZIKV infection in neural progenitor cells. Co-staining for Nestin (neural progenitor cells, green) and ZIKV E protein (red) was performed, and double-positive cells were quantified across six random fields, mean ± SD, ∗P < 0.05. Scale bar, 20 μm. (b) KEGG pathway enrichment bubble plot from <t>RNA-seq</t> <t>of</t> <t>HTR-8/Svneo</t> cells treated with CBNK-EVs (3 × 10 11 particles/mL, 10 μl) and ZIKV (MOI = 1) for 1 h. (c) GSEA analysis of transcriptomic data from HTR-8/Svneo cells. (d) Western blot analysis of ZIKV E and ZIKV NS5 protein levels after co-incubation of ZIKV particles with CBNK-EVs for the indicated time periods (0–30 min) prior to infection. (e) The abundance of NK cell-associated proteins in CBNK-EVs. ∗P < 0.05 (one-way ANOVA).
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Evaluation of the cytotoxicity and osteogenic properties of AgZ/GelSF coatings. (a) The cytotoxicity of extracts from coatings with different AgZ loadings on Ti discs was assessed using <t>the</t> <t>CCK-8</t> assay after incubation for 24 h. The amount of GelSF applied was 10 mg per disc. (b) The cytotoxicity of extracts from coatings with varying AgZ/GelSF fill amounts on Ti-6Al-4V cages was assessed using the CCK-8 assay after incubation for 24 h. Based on the results from panel (a), the Ag loading was fixed at 0.25 mg per 10 mg GelSF. AgZ/GelSF-38, 55, and 82 represent coating fill amounts of 38, 55, and 82 mg AgZ/GelSF per 10 g of cage, respectively. (c) Fluorescence images showing the viability of cells cultured in extracts from coatings with different AgZ/GelSF loadings. (d) Confocal microscopy images showing the growth status of cells cultured in various extracts. Green fluorescence indicates FITC-labeled actin filaments, while blue fluorescence indicates DAPI-stained nuclei. (e) Migration of cells cultured in material extracts for 12 and 24 h, visualized by fluorescence imaging. (f) ALP activity after 7 days of culture in extracts. (g) ARS staining after 14 days of culture, with insets showing macroscopic views of the staining. Based on the results from panel (b), the concentration of extracts used in panels (c-g) was 50%. (h) Schematic of rabbit femoral condyle bone defect model: the AgZ/GelSF coating was applied to a Ti-6Al-4V implant (Φ 6 mm × 9 mm) at a ratio of 55 mg per 10 g of implant, based on previous results. Osteogenic effects on the implant surface were evaluated after 6 weeks. (i) 3D and 2D reconstructed Micro-CT images showing bone tissue surrounding the implant. (j) Quantification of bone formation based on Micro-CT analysis, expressed as the ratio of bone volume to tissue volume. (k) Histological analysis of bone formation: top-view images of hard tissue sections from the femur stained with methylene blue/acid fuchsin. Yellow arrows indicate new bone formation on the implant surface. All data are presented as mean ± standard deviation from three independent samples.
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Evaluation of the cytotoxicity and osteogenic properties of AgZ/GelSF coatings. (a) The cytotoxicity of extracts from coatings with different AgZ loadings on Ti discs was assessed using <t>the</t> <t>CCK-8</t> assay after incubation for 24 h. The amount of GelSF applied was 10 mg per disc. (b) The cytotoxicity of extracts from coatings with varying AgZ/GelSF fill amounts on Ti-6Al-4V cages was assessed using the CCK-8 assay after incubation for 24 h. Based on the results from panel (a), the Ag loading was fixed at 0.25 mg per 10 mg GelSF. AgZ/GelSF-38, 55, and 82 represent coating fill amounts of 38, 55, and 82 mg AgZ/GelSF per 10 g of cage, respectively. (c) Fluorescence images showing the viability of cells cultured in extracts from coatings with different AgZ/GelSF loadings. (d) Confocal microscopy images showing the growth status of cells cultured in various extracts. Green fluorescence indicates FITC-labeled actin filaments, while blue fluorescence indicates DAPI-stained nuclei. (e) Migration of cells cultured in material extracts for 12 and 24 h, visualized by fluorescence imaging. (f) ALP activity after 7 days of culture in extracts. (g) ARS staining after 14 days of culture, with insets showing macroscopic views of the staining. Based on the results from panel (b), the concentration of extracts used in panels (c-g) was 50%. (h) Schematic of rabbit femoral condyle bone defect model: the AgZ/GelSF coating was applied to a Ti-6Al-4V implant (Φ 6 mm × 9 mm) at a ratio of 55 mg per 10 g of implant, based on previous results. Osteogenic effects on the implant surface were evaluated after 6 weeks. (i) 3D and 2D reconstructed Micro-CT images showing bone tissue surrounding the implant. (j) Quantification of bone formation based on Micro-CT analysis, expressed as the ratio of bone volume to tissue volume. (k) Histological analysis of bone formation: top-view images of hard tissue sections from the femur stained with methylene blue/acid fuchsin. Yellow arrows indicate new bone formation on the implant surface. All data are presented as mean ± standard deviation from three independent samples.
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CBNK-EVs rapidly impair ZIKV infectivity and protect host cells. (a) Multiplex immunohistochemical analysis of ZIKV infection in neural progenitor cells. Co-staining for Nestin (neural progenitor cells, green) and ZIKV E protein (red) was performed, and double-positive cells were quantified across six random fields, mean ± SD, ∗P < 0.05. Scale bar, 20 μm. (b) KEGG pathway enrichment bubble plot from RNA-seq of HTR-8/Svneo cells treated with CBNK-EVs (3 × 10 11 particles/mL, 10 μl) and ZIKV (MOI = 1) for 1 h. (c) GSEA analysis of transcriptomic data from HTR-8/Svneo cells. (d) Western blot analysis of ZIKV E and ZIKV NS5 protein levels after co-incubation of ZIKV particles with CBNK-EVs for the indicated time periods (0–30 min) prior to infection. (e) The abundance of NK cell-associated proteins in CBNK-EVs. ∗P < 0.05 (one-way ANOVA).

Journal: Bioactive Materials

Article Title: Cord blood natural killer cell-derived extracellular vesicles inhibit Zika virus infectivity through ITGB2/perforin-mediated envelope disruption in vitro and in vivo

doi: 10.1016/j.bioactmat.2026.01.030

Figure Lengend Snippet: CBNK-EVs rapidly impair ZIKV infectivity and protect host cells. (a) Multiplex immunohistochemical analysis of ZIKV infection in neural progenitor cells. Co-staining for Nestin (neural progenitor cells, green) and ZIKV E protein (red) was performed, and double-positive cells were quantified across six random fields, mean ± SD, ∗P < 0.05. Scale bar, 20 μm. (b) KEGG pathway enrichment bubble plot from RNA-seq of HTR-8/Svneo cells treated with CBNK-EVs (3 × 10 11 particles/mL, 10 μl) and ZIKV (MOI = 1) for 1 h. (c) GSEA analysis of transcriptomic data from HTR-8/Svneo cells. (d) Western blot analysis of ZIKV E and ZIKV NS5 protein levels after co-incubation of ZIKV particles with CBNK-EVs for the indicated time periods (0–30 min) prior to infection. (e) The abundance of NK cell-associated proteins in CBNK-EVs. ∗P < 0.05 (one-way ANOVA).

Article Snippet: Vero E6, BHK21, 293T and HTR-8/Svneo cells were purchased from ATCC.

Techniques: Infection, Multiplex Assay, Immunohistochemical staining, Staining, RNA Sequencing, Western Blot, Incubation

Evaluation of the cytotoxicity and osteogenic properties of AgZ/GelSF coatings. (a) The cytotoxicity of extracts from coatings with different AgZ loadings on Ti discs was assessed using the CCK-8 assay after incubation for 24 h. The amount of GelSF applied was 10 mg per disc. (b) The cytotoxicity of extracts from coatings with varying AgZ/GelSF fill amounts on Ti-6Al-4V cages was assessed using the CCK-8 assay after incubation for 24 h. Based on the results from panel (a), the Ag loading was fixed at 0.25 mg per 10 mg GelSF. AgZ/GelSF-38, 55, and 82 represent coating fill amounts of 38, 55, and 82 mg AgZ/GelSF per 10 g of cage, respectively. (c) Fluorescence images showing the viability of cells cultured in extracts from coatings with different AgZ/GelSF loadings. (d) Confocal microscopy images showing the growth status of cells cultured in various extracts. Green fluorescence indicates FITC-labeled actin filaments, while blue fluorescence indicates DAPI-stained nuclei. (e) Migration of cells cultured in material extracts for 12 and 24 h, visualized by fluorescence imaging. (f) ALP activity after 7 days of culture in extracts. (g) ARS staining after 14 days of culture, with insets showing macroscopic views of the staining. Based on the results from panel (b), the concentration of extracts used in panels (c-g) was 50%. (h) Schematic of rabbit femoral condyle bone defect model: the AgZ/GelSF coating was applied to a Ti-6Al-4V implant (Φ 6 mm × 9 mm) at a ratio of 55 mg per 10 g of implant, based on previous results. Osteogenic effects on the implant surface were evaluated after 6 weeks. (i) 3D and 2D reconstructed Micro-CT images showing bone tissue surrounding the implant. (j) Quantification of bone formation based on Micro-CT analysis, expressed as the ratio of bone volume to tissue volume. (k) Histological analysis of bone formation: top-view images of hard tissue sections from the femur stained with methylene blue/acid fuchsin. Yellow arrows indicate new bone formation on the implant surface. All data are presented as mean ± standard deviation from three independent samples.

Journal: Bioactive Materials

Article Title: Degradation-tunable coating with sustained silver release for spinal implants to prevent postoperative infections

doi: 10.1016/j.bioactmat.2026.02.035

Figure Lengend Snippet: Evaluation of the cytotoxicity and osteogenic properties of AgZ/GelSF coatings. (a) The cytotoxicity of extracts from coatings with different AgZ loadings on Ti discs was assessed using the CCK-8 assay after incubation for 24 h. The amount of GelSF applied was 10 mg per disc. (b) The cytotoxicity of extracts from coatings with varying AgZ/GelSF fill amounts on Ti-6Al-4V cages was assessed using the CCK-8 assay after incubation for 24 h. Based on the results from panel (a), the Ag loading was fixed at 0.25 mg per 10 mg GelSF. AgZ/GelSF-38, 55, and 82 represent coating fill amounts of 38, 55, and 82 mg AgZ/GelSF per 10 g of cage, respectively. (c) Fluorescence images showing the viability of cells cultured in extracts from coatings with different AgZ/GelSF loadings. (d) Confocal microscopy images showing the growth status of cells cultured in various extracts. Green fluorescence indicates FITC-labeled actin filaments, while blue fluorescence indicates DAPI-stained nuclei. (e) Migration of cells cultured in material extracts for 12 and 24 h, visualized by fluorescence imaging. (f) ALP activity after 7 days of culture in extracts. (g) ARS staining after 14 days of culture, with insets showing macroscopic views of the staining. Based on the results from panel (b), the concentration of extracts used in panels (c-g) was 50%. (h) Schematic of rabbit femoral condyle bone defect model: the AgZ/GelSF coating was applied to a Ti-6Al-4V implant (Φ 6 mm × 9 mm) at a ratio of 55 mg per 10 g of implant, based on previous results. Osteogenic effects on the implant surface were evaluated after 6 weeks. (i) 3D and 2D reconstructed Micro-CT images showing bone tissue surrounding the implant. (j) Quantification of bone formation based on Micro-CT analysis, expressed as the ratio of bone volume to tissue volume. (k) Histological analysis of bone formation: top-view images of hard tissue sections from the femur stained with methylene blue/acid fuchsin. Yellow arrows indicate new bone formation on the implant surface. All data are presented as mean ± standard deviation from three independent samples.

Article Snippet: Cell viability was assessed using a CCK-8 assay kit (HY-K0301, MedChemExpress, USA).

Techniques: CCK-8 Assay, Incubation, Fluorescence, Cell Culture, Confocal Microscopy, Labeling, Staining, Migration, Imaging, Activity Assay, Concentration Assay, Micro-CT, Standard Deviation