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nMSC@MT inheriting the original MSCs and could be effectively taken up by BMMSCs. a) Representative confocal images of PMVs with DiI-labeled cell membrane and Tht-labeled cytoplasm. Scale bar = 25 μm (above) and 1 μm (below). b) Illustration showing the co-extrusion procedure of nPMV and melatonin to obtain nMSC@MT. c) Representative TEM image of nMSC@MT. Scale bar = 100 nm. d, e) The sizes and the potential zetas of nPMV and nMSC@MT by DLS (n = 3). f, g) Coomassie brilliant blue and Western blot showing the preservation of stem cell markers, CD146, CD105, CD90, and functional protein, <t>CXCR4,</t> in PMV, nPMV, and nMSC@MT. h) DiI-labeled nMSC@MT (red) was detected in the cytoplasm of BMMSCs, suggesting the endocytosis of nMSC@MT. Scale bar = 10 μm. i) Immunofluorescence showing that a large amount of DiI-labeled nMSC@MT (red), released from the hydrogel, was taken up by CD146-labeled MSCs (green). Scale bar = 50 μm. j) BMMSCs uptake of DiI-labeled nMSC@MT in presence of blocking antibodies (anti-CXCR4) detected via flow cytometry. k) Analysis of the percentage of DiI positive cells in BMMSCs by flow cytometry (n = 3). l, m) Immunofluorescent staining images and corresponding semi-quantitative analysis (n = 3) of the uptake of DiI-labeled nPMV by BMMSCs in presence of blocking antibodies (anti-CXCR4). Scale bar = 50 μm. P-values are calculated using one-way ANOVA with Tukey's test, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, n.s. not significant ( b was created with bioRender. com).
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nMSC@MT inheriting the original MSCs and could be effectively taken up by BMMSCs. a) Representative confocal images of PMVs with DiI-labeled cell membrane and Tht-labeled cytoplasm. Scale bar = 25 μm (above) and 1 μm (below). b) Illustration showing the co-extrusion procedure of nPMV and melatonin to obtain nMSC@MT. c) Representative TEM image of nMSC@MT. Scale bar = 100 nm. d, e) The sizes and the potential zetas of nPMV and nMSC@MT by DLS (n = 3). f, g) Coomassie brilliant blue and Western blot showing the preservation of stem cell markers, CD146, CD105, CD90, and functional protein, <t>CXCR4,</t> in PMV, nPMV, and nMSC@MT. h) DiI-labeled nMSC@MT (red) was detected in the cytoplasm of BMMSCs, suggesting the endocytosis of nMSC@MT. Scale bar = 10 μm. i) Immunofluorescence showing that a large amount of DiI-labeled nMSC@MT (red), released from the hydrogel, was taken up by CD146-labeled MSCs (green). Scale bar = 50 μm. j) BMMSCs uptake of DiI-labeled nMSC@MT in presence of blocking antibodies (anti-CXCR4) detected via flow cytometry. k) Analysis of the percentage of DiI positive cells in BMMSCs by flow cytometry (n = 3). l, m) Immunofluorescent staining images and corresponding semi-quantitative analysis (n = 3) of the uptake of DiI-labeled nPMV by BMMSCs in presence of blocking antibodies (anti-CXCR4). Scale bar = 50 μm. P-values are calculated using one-way ANOVA with Tukey's test, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, n.s. not significant ( b was created with bioRender. com).
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nMSC@MT inheriting the original MSCs and could be effectively taken up by BMMSCs. a) Representative confocal images of PMVs with DiI-labeled cell membrane and Tht-labeled cytoplasm. Scale bar = 25 μm (above) and 1 μm (below). b) Illustration showing the co-extrusion procedure of nPMV and melatonin to obtain nMSC@MT. c) Representative TEM image of nMSC@MT. Scale bar = 100 nm. d, e) The sizes and the potential zetas of nPMV and nMSC@MT by DLS (n = 3). f, g) Coomassie brilliant blue and Western blot showing the preservation of stem cell markers, CD146, CD105, CD90, and functional protein, <t>CXCR4,</t> in PMV, nPMV, and nMSC@MT. h) DiI-labeled nMSC@MT (red) was detected in the cytoplasm of BMMSCs, suggesting the endocytosis of nMSC@MT. Scale bar = 10 μm. i) Immunofluorescence showing that a large amount of DiI-labeled nMSC@MT (red), released from the hydrogel, was taken up by CD146-labeled MSCs (green). Scale bar = 50 μm. j) BMMSCs uptake of DiI-labeled nMSC@MT in presence of blocking antibodies (anti-CXCR4) detected via flow cytometry. k) Analysis of the percentage of DiI positive cells in BMMSCs by flow cytometry (n = 3). l, m) Immunofluorescent staining images and corresponding semi-quantitative analysis (n = 3) of the uptake of DiI-labeled nPMV by BMMSCs in presence of blocking antibodies (anti-CXCR4). Scale bar = 50 μm. P-values are calculated using one-way ANOVA with Tukey's test, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, n.s. not significant ( b was created with bioRender. com).
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nMSC@MT inheriting the original MSCs and could be effectively taken up by BMMSCs. a) Representative confocal images of PMVs with DiI-labeled cell membrane and Tht-labeled cytoplasm. Scale bar = 25 μm (above) and 1 μm (below). b) Illustration showing the co-extrusion procedure of nPMV and melatonin to obtain nMSC@MT. c) Representative TEM image of nMSC@MT. Scale bar = 100 nm. d, e) The sizes and the potential zetas of nPMV and nMSC@MT by DLS (n = 3). f, g) Coomassie brilliant blue and Western blot showing the preservation of stem cell markers, CD146, CD105, CD90, and functional protein, <t>CXCR4,</t> in PMV, nPMV, and nMSC@MT. h) DiI-labeled nMSC@MT (red) was detected in the cytoplasm of BMMSCs, suggesting the endocytosis of nMSC@MT. Scale bar = 10 μm. i) Immunofluorescence showing that a large amount of DiI-labeled nMSC@MT (red), released from the hydrogel, was taken up by CD146-labeled MSCs (green). Scale bar = 50 μm. j) BMMSCs uptake of DiI-labeled nMSC@MT in presence of blocking antibodies (anti-CXCR4) detected via flow cytometry. k) Analysis of the percentage of DiI positive cells in BMMSCs by flow cytometry (n = 3). l, m) Immunofluorescent staining images and corresponding semi-quantitative analysis (n = 3) of the uptake of DiI-labeled nPMV by BMMSCs in presence of blocking antibodies (anti-CXCR4). Scale bar = 50 μm. P-values are calculated using one-way ANOVA with Tukey's test, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, n.s. not significant ( b was created with bioRender. com).
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nMSC@MT inheriting the original MSCs and could be effectively taken up by BMMSCs. a) Representative confocal images of PMVs with DiI-labeled cell membrane and Tht-labeled cytoplasm. Scale bar = 25 μm (above) and 1 μm (below). b) Illustration showing the co-extrusion procedure of nPMV and melatonin to obtain nMSC@MT. c) Representative TEM image of nMSC@MT. Scale bar = 100 nm. d, e) The sizes and the potential zetas of nPMV and nMSC@MT by DLS (n = 3). f, g) Coomassie brilliant blue and Western blot showing the preservation of stem cell markers, CD146, CD105, CD90, and functional protein, <t>CXCR4,</t> in PMV, nPMV, and nMSC@MT. h) DiI-labeled nMSC@MT (red) was detected in the cytoplasm of BMMSCs, suggesting the endocytosis of nMSC@MT. Scale bar = 10 μm. i) Immunofluorescence showing that a large amount of DiI-labeled nMSC@MT (red), released from the hydrogel, was taken up by CD146-labeled MSCs (green). Scale bar = 50 μm. j) BMMSCs uptake of DiI-labeled nMSC@MT in presence of blocking antibodies (anti-CXCR4) detected via flow cytometry. k) Analysis of the percentage of DiI positive cells in BMMSCs by flow cytometry (n = 3). l, m) Immunofluorescent staining images and corresponding semi-quantitative analysis (n = 3) of the uptake of DiI-labeled nPMV by BMMSCs in presence of blocking antibodies (anti-CXCR4). Scale bar = 50 μm. P-values are calculated using one-way ANOVA with Tukey's test, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, n.s. not significant ( b was created with bioRender. com).
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nMSC@MT inheriting the original MSCs and could be effectively taken up by BMMSCs. a) Representative confocal images of PMVs with DiI-labeled cell membrane and Tht-labeled cytoplasm. Scale bar = 25 μm (above) and 1 μm (below). b) Illustration showing the co-extrusion procedure of nPMV and melatonin to obtain nMSC@MT. c) Representative TEM image of nMSC@MT. Scale bar = 100 nm. d, e) The sizes and the potential zetas of nPMV and nMSC@MT by DLS (n = 3). f, g) Coomassie brilliant blue and Western blot showing the preservation of stem cell markers, CD146, CD105, CD90, and functional protein, CXCR4, in PMV, nPMV, and nMSC@MT. h) DiI-labeled nMSC@MT (red) was detected in the cytoplasm of BMMSCs, suggesting the endocytosis of nMSC@MT. Scale bar = 10 μm. i) Immunofluorescence showing that a large amount of DiI-labeled nMSC@MT (red), released from the hydrogel, was taken up by CD146-labeled MSCs (green). Scale bar = 50 μm. j) BMMSCs uptake of DiI-labeled nMSC@MT in presence of blocking antibodies (anti-CXCR4) detected via flow cytometry. k) Analysis of the percentage of DiI positive cells in BMMSCs by flow cytometry (n = 3). l, m) Immunofluorescent staining images and corresponding semi-quantitative analysis (n = 3) of the uptake of DiI-labeled nPMV by BMMSCs in presence of blocking antibodies (anti-CXCR4). Scale bar = 50 μm. P-values are calculated using one-way ANOVA with Tukey's test, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, n.s. not significant ( b was created with bioRender. com).

Journal: Bioactive Materials

Article Title: MSC-mimicking nanovesicle embedded bio-adhesive hydrogel for dual immunomodulation and osteogenesis to promote maxillofacial bone regeneration

doi: 10.1016/j.bioactmat.2026.02.032

Figure Lengend Snippet: nMSC@MT inheriting the original MSCs and could be effectively taken up by BMMSCs. a) Representative confocal images of PMVs with DiI-labeled cell membrane and Tht-labeled cytoplasm. Scale bar = 25 μm (above) and 1 μm (below). b) Illustration showing the co-extrusion procedure of nPMV and melatonin to obtain nMSC@MT. c) Representative TEM image of nMSC@MT. Scale bar = 100 nm. d, e) The sizes and the potential zetas of nPMV and nMSC@MT by DLS (n = 3). f, g) Coomassie brilliant blue and Western blot showing the preservation of stem cell markers, CD146, CD105, CD90, and functional protein, CXCR4, in PMV, nPMV, and nMSC@MT. h) DiI-labeled nMSC@MT (red) was detected in the cytoplasm of BMMSCs, suggesting the endocytosis of nMSC@MT. Scale bar = 10 μm. i) Immunofluorescence showing that a large amount of DiI-labeled nMSC@MT (red), released from the hydrogel, was taken up by CD146-labeled MSCs (green). Scale bar = 50 μm. j) BMMSCs uptake of DiI-labeled nMSC@MT in presence of blocking antibodies (anti-CXCR4) detected via flow cytometry. k) Analysis of the percentage of DiI positive cells in BMMSCs by flow cytometry (n = 3). l, m) Immunofluorescent staining images and corresponding semi-quantitative analysis (n = 3) of the uptake of DiI-labeled nPMV by BMMSCs in presence of blocking antibodies (anti-CXCR4). Scale bar = 50 μm. P-values are calculated using one-way ANOVA with Tukey's test, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, n.s. not significant ( b was created with bioRender. com).

Article Snippet: To evaluate the involvement of CXCR4 in the uptake of nPMV, prior to co-culture with BMMSCs, nPMV was pre-treated with 160 nM CXCR4 blocking antibody (Ulocuplumab, Cat. HY-P99272, MedChemExpress, China) at room temperature for 1 h. Then nPMV was introduced into BMMSC cultures and incubated for 24 h. Subsequent analyses included flow cytometric analysis and immunofluorescence staining. nPMV without blocking antibody was served as a control.

Techniques: Labeling, Membrane, Western Blot, Preserving, Functional Assay, Immunofluorescence, Blocking Assay, Flow Cytometry, Staining