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ZIF-8/sEVs alleviate the inhibitory effects of the M1 M ϕ -mediated inflammatory microenvironment on BMSC migration and chondrogenic differentiation. A) Representative immunofluorescence images of the chemotaxis-associated proteins <t>CXCL12</t> (green) and CXCR4 (red) in BMSCs cultured under different conditioned media. Nuclei were stained with DAPI (blue). B) Representative optical microscopy images of the scratch wound healing assay at 0 h and 24 h to assess the migratory capacity of BMSCs. C, D) Semi-quantitative analysis of the relative fluorescence intensity of CXCL12 and CXCR4 (C) and the wound healing ratio (D), indicating that ZIF-8/sEVs restore migration inhibited by M1 CM. E–G) Representative immunofluorescence staining of key chondrogenic markers in BMSCs, including (E) Type II Collagen (COL II, red), (F) Aggrecan (red), and (G) Sox-9 (red). The cytoskeleton was stained with Phalloidin (green) and nuclei with DAPI (blue). H–J) Semi-quantitative analysis of the relative fluorescence intensity for (H) COL II, (I) Aggrecan, and (J) Sox-9. M1 CM: M1 M ϕ conditioned media. n = 3. ∗∗p < 0.01, ∗∗∗p < 0.001, ns: p-value with no significance. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)
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ZIF-8/sEVs alleviate the inhibitory effects of the M1 M ϕ -mediated inflammatory microenvironment on BMSC migration and chondrogenic differentiation. A) Representative immunofluorescence images of the chemotaxis-associated proteins <t>CXCL12</t> (green) and CXCR4 (red) in BMSCs cultured under different conditioned media. Nuclei were stained with DAPI (blue). B) Representative optical microscopy images of the scratch wound healing assay at 0 h and 24 h to assess the migratory capacity of BMSCs. C, D) Semi-quantitative analysis of the relative fluorescence intensity of CXCL12 and CXCR4 (C) and the wound healing ratio (D), indicating that ZIF-8/sEVs restore migration inhibited by M1 CM. E–G) Representative immunofluorescence staining of key chondrogenic markers in BMSCs, including (E) Type II Collagen (COL II, red), (F) Aggrecan (red), and (G) Sox-9 (red). The cytoskeleton was stained with Phalloidin (green) and nuclei with DAPI (blue). H–J) Semi-quantitative analysis of the relative fluorescence intensity for (H) COL II, (I) Aggrecan, and (J) Sox-9. M1 CM: M1 M ϕ conditioned media. n = 3. ∗∗p < 0.01, ∗∗∗p < 0.001, ns: p-value with no significance. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)
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ZIF-8/sEVs alleviate the inhibitory effects of the M1 M ϕ -mediated inflammatory microenvironment on BMSC migration and chondrogenic differentiation. A) Representative immunofluorescence images of the chemotaxis-associated proteins <t>CXCL12</t> (green) and CXCR4 (red) in BMSCs cultured under different conditioned media. Nuclei were stained with DAPI (blue). B) Representative optical microscopy images of the scratch wound healing assay at 0 h and 24 h to assess the migratory capacity of BMSCs. C, D) Semi-quantitative analysis of the relative fluorescence intensity of CXCL12 and CXCR4 (C) and the wound healing ratio (D), indicating that ZIF-8/sEVs restore migration inhibited by M1 CM. E–G) Representative immunofluorescence staining of key chondrogenic markers in BMSCs, including (E) Type II Collagen (COL II, red), (F) Aggrecan (red), and (G) Sox-9 (red). The cytoskeleton was stained with Phalloidin (green) and nuclei with DAPI (blue). H–J) Semi-quantitative analysis of the relative fluorescence intensity for (H) COL II, (I) Aggrecan, and (J) Sox-9. M1 CM: M1 M ϕ conditioned media. n = 3. ∗∗p < 0.01, ∗∗∗p < 0.001, ns: p-value with no significance. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)
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ZIF-8/sEVs alleviate the inhibitory effects of the M1 M ϕ -mediated inflammatory microenvironment on BMSC migration and chondrogenic differentiation. A) Representative immunofluorescence images of the chemotaxis-associated proteins <t>CXCL12</t> (green) and CXCR4 (red) in BMSCs cultured under different conditioned media. Nuclei were stained with DAPI (blue). B) Representative optical microscopy images of the scratch wound healing assay at 0 h and 24 h to assess the migratory capacity of BMSCs. C, D) Semi-quantitative analysis of the relative fluorescence intensity of CXCL12 and CXCR4 (C) and the wound healing ratio (D), indicating that ZIF-8/sEVs restore migration inhibited by M1 CM. E–G) Representative immunofluorescence staining of key chondrogenic markers in BMSCs, including (E) Type II Collagen (COL II, red), (F) Aggrecan (red), and (G) Sox-9 (red). The cytoskeleton was stained with Phalloidin (green) and nuclei with DAPI (blue). H–J) Semi-quantitative analysis of the relative fluorescence intensity for (H) COL II, (I) Aggrecan, and (J) Sox-9. M1 CM: M1 M ϕ conditioned media. n = 3. ∗∗p < 0.01, ∗∗∗p < 0.001, ns: p-value with no significance. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)
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ZIF-8/sEVs alleviate the inhibitory effects of the M1 M ϕ -mediated inflammatory microenvironment on BMSC migration and chondrogenic differentiation. A) Representative immunofluorescence images of the chemotaxis-associated proteins <t>CXCL12</t> (green) and CXCR4 (red) in BMSCs cultured under different conditioned media. Nuclei were stained with DAPI (blue). B) Representative optical microscopy images of the scratch wound healing assay at 0 h and 24 h to assess the migratory capacity of BMSCs. C, D) Semi-quantitative analysis of the relative fluorescence intensity of CXCL12 and CXCR4 (C) and the wound healing ratio (D), indicating that ZIF-8/sEVs restore migration inhibited by M1 CM. E–G) Representative immunofluorescence staining of key chondrogenic markers in BMSCs, including (E) Type II Collagen (COL II, red), (F) Aggrecan (red), and (G) Sox-9 (red). The cytoskeleton was stained with Phalloidin (green) and nuclei with DAPI (blue). H–J) Semi-quantitative analysis of the relative fluorescence intensity for (H) COL II, (I) Aggrecan, and (J) Sox-9. M1 CM: M1 M ϕ conditioned media. n = 3. ∗∗p < 0.01, ∗∗∗p < 0.001, ns: p-value with no significance. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)
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ZIF-8/sEVs alleviate the inhibitory effects of the M1 M ϕ -mediated inflammatory microenvironment on BMSC migration and chondrogenic differentiation. A) Representative immunofluorescence images of the chemotaxis-associated proteins <t>CXCL12</t> (green) and CXCR4 (red) in BMSCs cultured under different conditioned media. Nuclei were stained with DAPI (blue). B) Representative optical microscopy images of the scratch wound healing assay at 0 h and 24 h to assess the migratory capacity of BMSCs. C, D) Semi-quantitative analysis of the relative fluorescence intensity of CXCL12 and CXCR4 (C) and the wound healing ratio (D), indicating that ZIF-8/sEVs restore migration inhibited by M1 CM. E–G) Representative immunofluorescence staining of key chondrogenic markers in BMSCs, including (E) Type II Collagen (COL II, red), (F) Aggrecan (red), and (G) Sox-9 (red). The cytoskeleton was stained with Phalloidin (green) and nuclei with DAPI (blue). H–J) Semi-quantitative analysis of the relative fluorescence intensity for (H) COL II, (I) Aggrecan, and (J) Sox-9. M1 CM: M1 M ϕ conditioned media. n = 3. ∗∗p < 0.01, ∗∗∗p < 0.001, ns: p-value with no significance. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)
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ZIF-8/sEVs alleviate the inhibitory effects of the M1 M ϕ -mediated inflammatory microenvironment on BMSC migration and chondrogenic differentiation. A) Representative immunofluorescence images of the chemotaxis-associated proteins <t>CXCL12</t> (green) and CXCR4 (red) in BMSCs cultured under different conditioned media. Nuclei were stained with DAPI (blue). B) Representative optical microscopy images of the scratch wound healing assay at 0 h and 24 h to assess the migratory capacity of BMSCs. C, D) Semi-quantitative analysis of the relative fluorescence intensity of CXCL12 and CXCR4 (C) and the wound healing ratio (D), indicating that ZIF-8/sEVs restore migration inhibited by M1 CM. E–G) Representative immunofluorescence staining of key chondrogenic markers in BMSCs, including (E) Type II Collagen (COL II, red), (F) Aggrecan (red), and (G) Sox-9 (red). The cytoskeleton was stained with Phalloidin (green) and nuclei with DAPI (blue). H–J) Semi-quantitative analysis of the relative fluorescence intensity for (H) COL II, (I) Aggrecan, and (J) Sox-9. M1 CM: M1 M ϕ conditioned media. n = 3. ∗∗p < 0.01, ∗∗∗p < 0.001, ns: p-value with no significance. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)
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ZIF-8/sEVs alleviate the inhibitory effects of the M1 M ϕ -mediated inflammatory microenvironment on BMSC migration and chondrogenic differentiation. A) Representative immunofluorescence images of the chemotaxis-associated proteins CXCL12 (green) and CXCR4 (red) in BMSCs cultured under different conditioned media. Nuclei were stained with DAPI (blue). B) Representative optical microscopy images of the scratch wound healing assay at 0 h and 24 h to assess the migratory capacity of BMSCs. C, D) Semi-quantitative analysis of the relative fluorescence intensity of CXCL12 and CXCR4 (C) and the wound healing ratio (D), indicating that ZIF-8/sEVs restore migration inhibited by M1 CM. E–G) Representative immunofluorescence staining of key chondrogenic markers in BMSCs, including (E) Type II Collagen (COL II, red), (F) Aggrecan (red), and (G) Sox-9 (red). The cytoskeleton was stained with Phalloidin (green) and nuclei with DAPI (blue). H–J) Semi-quantitative analysis of the relative fluorescence intensity for (H) COL II, (I) Aggrecan, and (J) Sox-9. M1 CM: M1 M ϕ conditioned media. n = 3. ∗∗p < 0.01, ∗∗∗p < 0.001, ns: p-value with no significance. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Journal: Materials Today Bio

Article Title: Orchestrating the immuno-chondrogenic microenvironment via a bio-inorganic hybrid microsphere system for tendon-to-bone healing

doi: 10.1016/j.mtbio.2026.102851

Figure Lengend Snippet: ZIF-8/sEVs alleviate the inhibitory effects of the M1 M ϕ -mediated inflammatory microenvironment on BMSC migration and chondrogenic differentiation. A) Representative immunofluorescence images of the chemotaxis-associated proteins CXCL12 (green) and CXCR4 (red) in BMSCs cultured under different conditioned media. Nuclei were stained with DAPI (blue). B) Representative optical microscopy images of the scratch wound healing assay at 0 h and 24 h to assess the migratory capacity of BMSCs. C, D) Semi-quantitative analysis of the relative fluorescence intensity of CXCL12 and CXCR4 (C) and the wound healing ratio (D), indicating that ZIF-8/sEVs restore migration inhibited by M1 CM. E–G) Representative immunofluorescence staining of key chondrogenic markers in BMSCs, including (E) Type II Collagen (COL II, red), (F) Aggrecan (red), and (G) Sox-9 (red). The cytoskeleton was stained with Phalloidin (green) and nuclei with DAPI (blue). H–J) Semi-quantitative analysis of the relative fluorescence intensity for (H) COL II, (I) Aggrecan, and (J) Sox-9. M1 CM: M1 M ϕ conditioned media. n = 3. ∗∗p < 0.01, ∗∗∗p < 0.001, ns: p-value with no significance. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Antibodies against HSP70, CD9, CD81, Calnexin, iNOS, CD206, TNF-α, IL-10, CD29, CD90, CD68, CXCL12, CXCR4, COL II, Aggrecan, Sox-9, PI3K, p-PI3K, AKT, p-AKT, and GAPDH were purchased from Abcam (UK), Cell Signaling Technology (USA), Servicebio (Wuhan, China), Abclonal (Wuhan, China), Biolegend (Beijing, China), or Proteintech (Wuhan, China) (detailed information listed in ).

Techniques: Migration, Immunofluorescence, Chemotaxis Assay, Cell Culture, Staining, Microscopy, Wound Healing Assay, Fluorescence