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    Beyotime dapi anti fluorescence quenching mounting
    miR-423–5p was identified as a key osteogenic factor in sEVs. (A) Schematic illustration of the experimental design used to assess the impact of miR-423–5p on PDLC proliferation and osteogenic differentiation. n = 3. ( B ) EdU assay evaluating the influence of miR-423–5p on PDLC proliferation. Green fluorescence indicates EdU-labeled cells, and blue fluorescence indicates <t>DAPI-stained</t> nuclei. Scale bar = 100 μm. ( C ) Quantitative assessment of EdU-positive cell proportions. n = 5. ( D ) CCK-8 experiment evaluating the impact of miR-423–5p on PDLC proliferation. n = 6. ( E ) ALP and ARS assays assessing the impact of miR-423–5p on PDLC osteogenic differentiation. Scale bar = 200 μm. ( F ) Quantitative analysis of ALP activity. n = 6. ( G ) Western blot analysis of the influence of miR-423–5p on the osteogenic proteins expression (COL1, Runx2) in PDLCs. ( H ) Quantitative analysis of protein expression. n = 3. ( I ) The influence of miR-423–5p on the expression of osteogenic genes COL1 and Runx2 in PDLCs, evaluated using a PCR assay. n = 3. ( J ) Venn diagram showing the target genes predicted by miRTarBase, miRWalk, and TargetScan. ( K ) Pathway interaction map of the intersecting target genes. ( L ) ALP and ARS assays assessing the impact of PLCB1 on osteogenic differentiation in PDLCs. Scale bar = 200 μm. ( M ) Quantitative analysis of ALP activity. n = 6. ( N ) ALP staining and quantitative analysis of ALP activity in PDLCs co-transfected with miR-423–5p mimic and PLCB1 overexpression plasmid (n = 6). ( O ) Effects of miR-423–5p mimic combined with PLCB1 overexpression on the expression levels of PLCB1, Wnt3, and β-catenin in PDLCs.
    Dapi Anti Fluorescence Quenching Mounting, supplied by Beyotime, used in various techniques. Bioz Stars score: 99/100, based on 32382 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/dapi+anti+fluorescence+quenching+mounting/DAPI/pmc12703869-275-5-9
    Average 99 stars, based on 32382 article reviews
    dapi anti fluorescence quenching mounting - by Bioz Stars, 2026-10
    99/100 stars

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    1) Product Images from "miR-423-5p-enriched small extracellular vesicles drive periodontal regeneration via Sfrp2+ cell expansion"

    Article Title: miR-423-5p-enriched small extracellular vesicles drive periodontal regeneration via Sfrp2+ cell expansion

    Journal: Bioactive Materials

    doi: 10.1016/j.bioactmat.2025.11.026

    miR-423–5p was identified as a key osteogenic factor in sEVs. (A) Schematic illustration of the experimental design used to assess the impact of miR-423–5p on PDLC proliferation and osteogenic differentiation. n = 3. ( B ) EdU assay evaluating the influence of miR-423–5p on PDLC proliferation. Green fluorescence indicates EdU-labeled cells, and blue fluorescence indicates DAPI-stained nuclei. Scale bar = 100 μm. ( C ) Quantitative assessment of EdU-positive cell proportions. n = 5. ( D ) CCK-8 experiment evaluating the impact of miR-423–5p on PDLC proliferation. n = 6. ( E ) ALP and ARS assays assessing the impact of miR-423–5p on PDLC osteogenic differentiation. Scale bar = 200 μm. ( F ) Quantitative analysis of ALP activity. n = 6. ( G ) Western blot analysis of the influence of miR-423–5p on the osteogenic proteins expression (COL1, Runx2) in PDLCs. ( H ) Quantitative analysis of protein expression. n = 3. ( I ) The influence of miR-423–5p on the expression of osteogenic genes COL1 and Runx2 in PDLCs, evaluated using a PCR assay. n = 3. ( J ) Venn diagram showing the target genes predicted by miRTarBase, miRWalk, and TargetScan. ( K ) Pathway interaction map of the intersecting target genes. ( L ) ALP and ARS assays assessing the impact of PLCB1 on osteogenic differentiation in PDLCs. Scale bar = 200 μm. ( M ) Quantitative analysis of ALP activity. n = 6. ( N ) ALP staining and quantitative analysis of ALP activity in PDLCs co-transfected with miR-423–5p mimic and PLCB1 overexpression plasmid (n = 6). ( O ) Effects of miR-423–5p mimic combined with PLCB1 overexpression on the expression levels of PLCB1, Wnt3, and β-catenin in PDLCs.
    Figure Legend Snippet: miR-423–5p was identified as a key osteogenic factor in sEVs. (A) Schematic illustration of the experimental design used to assess the impact of miR-423–5p on PDLC proliferation and osteogenic differentiation. n = 3. ( B ) EdU assay evaluating the influence of miR-423–5p on PDLC proliferation. Green fluorescence indicates EdU-labeled cells, and blue fluorescence indicates DAPI-stained nuclei. Scale bar = 100 μm. ( C ) Quantitative assessment of EdU-positive cell proportions. n = 5. ( D ) CCK-8 experiment evaluating the impact of miR-423–5p on PDLC proliferation. n = 6. ( E ) ALP and ARS assays assessing the impact of miR-423–5p on PDLC osteogenic differentiation. Scale bar = 200 μm. ( F ) Quantitative analysis of ALP activity. n = 6. ( G ) Western blot analysis of the influence of miR-423–5p on the osteogenic proteins expression (COL1, Runx2) in PDLCs. ( H ) Quantitative analysis of protein expression. n = 3. ( I ) The influence of miR-423–5p on the expression of osteogenic genes COL1 and Runx2 in PDLCs, evaluated using a PCR assay. n = 3. ( J ) Venn diagram showing the target genes predicted by miRTarBase, miRWalk, and TargetScan. ( K ) Pathway interaction map of the intersecting target genes. ( L ) ALP and ARS assays assessing the impact of PLCB1 on osteogenic differentiation in PDLCs. Scale bar = 200 μm. ( M ) Quantitative analysis of ALP activity. n = 6. ( N ) ALP staining and quantitative analysis of ALP activity in PDLCs co-transfected with miR-423–5p mimic and PLCB1 overexpression plasmid (n = 6). ( O ) Effects of miR-423–5p mimic combined with PLCB1 overexpression on the expression levels of PLCB1, Wnt3, and β-catenin in PDLCs.

    Techniques Used: EdU Assay, Fluorescence, Labeling, Staining, CCK-8 Assay, Activity Assay, Western Blot, Expressing, Transfection, Over Expression, Plasmid Preparation

    sEVs miR−423−5p increased the osteogenic differentiation potential of sEVs. (A) TEM characterization of the morphology of sEVs and sEVs miR−423−5p morphology. Scale bar = 200 nm. ( B ) Nanoflow cytometry of the particle size distribution of sEVs and sEVs miR−423−5p . ( C ) Western blot analysis of sEVs and sEVs miR−423−5p markers, including CD9, CD63, HSP90, TSG101, and the endoplasmic reticulum protein calnexin. ( D ) Analysis of the Zeta potential of sEVs and sEVs miR−423−5p . n = 3. ( E ) Quantitative PCR analysis comparing miR-423–5p expression levels between sEVs and sEVs miR−423−5p . n = 3. ( F ) Confocal microscopy imaging of sEV/sEVs miR−423−5p uptake by PDLCs. PDLC nuclei are marked with DAPI (blue), sEVs stained with PKH26 (red dots), the cytoskeleton stained with Cell Mask Actin (red filaments), and miR-423–5p is depicted in green. Scale bar = 20 μm. ( G ) Quantitative PCR assessment of miR-423–5p, miR-100–5p, miR-125b-5p, miR-26a-5p, and miR-24–3p levels in PDLCs following sEV/sEVs miR−423−5p uptake. n = 3. ( H ) RIP-qPCR analysis showing that miR-423–5p delivered by sEVs miR−423−5p is loaded onto Ago2 (Argonaute 2) protein in PDLCs. n = 3. ( I ) CCK-8 experiment evaluating the impact of sEVs miR−423−5p on PDLC proliferation. n = 4. ( J ) Western blot analysis of proliferative markers Ki67, MCM7, and PCNA (proliferating cell nuclear antigen) in sEVs miR−423−5p -treated PDLCs. ( K ) Quantitative analysis of protein expression. n = 3. ( L ) ALP and ARS assays assessing the influence of sEVs miR−423−5p on mediating PDLC osteogenic differentiation. Scale bar = 200 μm. ( M ) Quantitative analysis of ALP activity. n = 6. ( N ) PCR analysis of osteogenic gene expression in PDLCs following sEVs miR−423−5p treatment. n = 3. ( O ) Western blot quantification of osteogenic protein levels in sEVs miR−423−5p -treated PDLCs. ( P ) Quantitative analysis of protein expression. n = 3.
    Figure Legend Snippet: sEVs miR−423−5p increased the osteogenic differentiation potential of sEVs. (A) TEM characterization of the morphology of sEVs and sEVs miR−423−5p morphology. Scale bar = 200 nm. ( B ) Nanoflow cytometry of the particle size distribution of sEVs and sEVs miR−423−5p . ( C ) Western blot analysis of sEVs and sEVs miR−423−5p markers, including CD9, CD63, HSP90, TSG101, and the endoplasmic reticulum protein calnexin. ( D ) Analysis of the Zeta potential of sEVs and sEVs miR−423−5p . n = 3. ( E ) Quantitative PCR analysis comparing miR-423–5p expression levels between sEVs and sEVs miR−423−5p . n = 3. ( F ) Confocal microscopy imaging of sEV/sEVs miR−423−5p uptake by PDLCs. PDLC nuclei are marked with DAPI (blue), sEVs stained with PKH26 (red dots), the cytoskeleton stained with Cell Mask Actin (red filaments), and miR-423–5p is depicted in green. Scale bar = 20 μm. ( G ) Quantitative PCR assessment of miR-423–5p, miR-100–5p, miR-125b-5p, miR-26a-5p, and miR-24–3p levels in PDLCs following sEV/sEVs miR−423−5p uptake. n = 3. ( H ) RIP-qPCR analysis showing that miR-423–5p delivered by sEVs miR−423−5p is loaded onto Ago2 (Argonaute 2) protein in PDLCs. n = 3. ( I ) CCK-8 experiment evaluating the impact of sEVs miR−423−5p on PDLC proliferation. n = 4. ( J ) Western blot analysis of proliferative markers Ki67, MCM7, and PCNA (proliferating cell nuclear antigen) in sEVs miR−423−5p -treated PDLCs. ( K ) Quantitative analysis of protein expression. n = 3. ( L ) ALP and ARS assays assessing the influence of sEVs miR−423−5p on mediating PDLC osteogenic differentiation. Scale bar = 200 μm. ( M ) Quantitative analysis of ALP activity. n = 6. ( N ) PCR analysis of osteogenic gene expression in PDLCs following sEVs miR−423−5p treatment. n = 3. ( O ) Western blot quantification of osteogenic protein levels in sEVs miR−423−5p -treated PDLCs. ( P ) Quantitative analysis of protein expression. n = 3.

    Techniques Used: Cytometry, Western Blot, Zeta Potential Analyzer, Real-time Polymerase Chain Reaction, Expressing, Confocal Microscopy, Imaging, Staining, CCK-8 Assay, Activity Assay, Gene Expression

    sEVs miR−423−5p promoted periodontal regeneration in rats. ( A ) Morphological characterization of collagen sponges and sEV/sEVs miR−423−5p -loaded collagen sponges via SEM. White arrows indicate sEVs and sEVs miR−423−5p . Scale bars: 2 μm, and 100 nm at increasing magnifications. ( B ) Analysis of the release rate of sEVs/sEVs miR−423−5p from collagen sponges. ( C ) Schematic representation of the periodontal defect model. ( D ) Confocal imaging revealed the distribution of sEVs and sEVs miR−423−5p within the periodontal defects in rats. PKH26-labeled sEVs are shown in red (red arrows), miR-423–5p in green (green arrows), colocalization of sEVs and miR-423–5p in orange (orange arrows), and DAPI-stained nuclei in blue. Scale bars: 500 μm, 100 μm, 50 μm, and 20 μm at increasing magnification. ( E ) CT images of periodontal defect regions in rats. Scale bar = 1 mm. ( F ) Quantitative analysis of new bone volume fraction (bone volume/tissue volume; BV/TV), bone mineral density (BMD), trabecular thickness (Tb. Th), and trabecular separation (Tb. Sp) in the defect areas. n = 4. ( G ) HE and Masson's trichrome staining of regenerated periodontal tissue, with black dashed lines indicating defect regions and red asterisks highlighting periodontal ligament-like structures. Scale bars: 625 μm (low magnification), 100 μm (high magnification). AB: alveolar bone; CS: collagen sponges; PDL: periodontal ligament; d: dentin.
    Figure Legend Snippet: sEVs miR−423−5p promoted periodontal regeneration in rats. ( A ) Morphological characterization of collagen sponges and sEV/sEVs miR−423−5p -loaded collagen sponges via SEM. White arrows indicate sEVs and sEVs miR−423−5p . Scale bars: 2 μm, and 100 nm at increasing magnifications. ( B ) Analysis of the release rate of sEVs/sEVs miR−423−5p from collagen sponges. ( C ) Schematic representation of the periodontal defect model. ( D ) Confocal imaging revealed the distribution of sEVs and sEVs miR−423−5p within the periodontal defects in rats. PKH26-labeled sEVs are shown in red (red arrows), miR-423–5p in green (green arrows), colocalization of sEVs and miR-423–5p in orange (orange arrows), and DAPI-stained nuclei in blue. Scale bars: 500 μm, 100 μm, 50 μm, and 20 μm at increasing magnification. ( E ) CT images of periodontal defect regions in rats. Scale bar = 1 mm. ( F ) Quantitative analysis of new bone volume fraction (bone volume/tissue volume; BV/TV), bone mineral density (BMD), trabecular thickness (Tb. Th), and trabecular separation (Tb. Sp) in the defect areas. n = 4. ( G ) HE and Masson's trichrome staining of regenerated periodontal tissue, with black dashed lines indicating defect regions and red asterisks highlighting periodontal ligament-like structures. Scale bars: 625 μm (low magnification), 100 μm (high magnification). AB: alveolar bone; CS: collagen sponges; PDL: periodontal ligament; d: dentin.

    Techniques Used: Imaging, Labeling, Staining

    Related Articles

    EdU Assay:

    Article Title: miR-423-5p-enriched small extracellular vesicles drive periodontal regeneration via Sfrp2+ cell expansion
    Article Snippet: For immunofluorescence staining, heart, liver, spleen, lung, kidney, and mandible samples were frozen in optimal cutting temperature compound (O.C.T. compound, Sakura Finetek, USA) and sectioned (5 μm).For immunofluorescence staining, heart, liver, spleen, lung, kidney, and mandible samples were frozen in optimal cutting temperature compound (O.C.T. compound, Sakura Finetek, USA) and sectioned (5 μm).. The slides were mounted with DAPI anti-fluorescence quenching mounting (Beyotime, Shanghai, China).. Observations and image collection were performed using a confocal microscope.Observations and image collection were performed using a confocal microscope.

    Fluorescence:

    Article Title: miR-423-5p-enriched small extracellular vesicles drive periodontal regeneration via Sfrp2+ cell expansion
    Article Snippet: For immunofluorescence staining, heart, liver, spleen, lung, kidney, and mandible samples were frozen in optimal cutting temperature compound (O.C.T. compound, Sakura Finetek, USA) and sectioned (5 μm).For immunofluorescence staining, heart, liver, spleen, lung, kidney, and mandible samples were frozen in optimal cutting temperature compound (O.C.T. compound, Sakura Finetek, USA) and sectioned (5 μm).. The slides were mounted with DAPI anti-fluorescence quenching mounting (Beyotime, Shanghai, China).. Observations and image collection were performed using a confocal microscope.Observations and image collection were performed using a confocal microscope.

    Labeling:

    Article Title: miR-423-5p-enriched small extracellular vesicles drive periodontal regeneration via Sfrp2+ cell expansion
    Article Snippet: For immunofluorescence staining, heart, liver, spleen, lung, kidney, and mandible samples were frozen in optimal cutting temperature compound (O.C.T. compound, Sakura Finetek, USA) and sectioned (5 μm).For immunofluorescence staining, heart, liver, spleen, lung, kidney, and mandible samples were frozen in optimal cutting temperature compound (O.C.T. compound, Sakura Finetek, USA) and sectioned (5 μm).. The slides were mounted with DAPI anti-fluorescence quenching mounting (Beyotime, Shanghai, China).. Observations and image collection were performed using a confocal microscope.Observations and image collection were performed using a confocal microscope.

    Staining:

    Article Title: miR-423-5p-enriched small extracellular vesicles drive periodontal regeneration via Sfrp2+ cell expansion
    Article Snippet: For immunofluorescence staining, heart, liver, spleen, lung, kidney, and mandible samples were frozen in optimal cutting temperature compound (O.C.T. compound, Sakura Finetek, USA) and sectioned (5 μm).For immunofluorescence staining, heart, liver, spleen, lung, kidney, and mandible samples were frozen in optimal cutting temperature compound (O.C.T. compound, Sakura Finetek, USA) and sectioned (5 μm).. The slides were mounted with DAPI anti-fluorescence quenching mounting (Beyotime, Shanghai, China).. Observations and image collection were performed using a confocal microscope.Observations and image collection were performed using a confocal microscope.

    CCK-8 Assay:

    Article Title: miR-423-5p-enriched small extracellular vesicles drive periodontal regeneration via Sfrp2+ cell expansion
    Article Snippet: For immunofluorescence staining, heart, liver, spleen, lung, kidney, and mandible samples were frozen in optimal cutting temperature compound (O.C.T. compound, Sakura Finetek, USA) and sectioned (5 μm).For immunofluorescence staining, heart, liver, spleen, lung, kidney, and mandible samples were frozen in optimal cutting temperature compound (O.C.T. compound, Sakura Finetek, USA) and sectioned (5 μm).. The slides were mounted with DAPI anti-fluorescence quenching mounting (Beyotime, Shanghai, China).. Observations and image collection were performed using a confocal microscope.Observations and image collection were performed using a confocal microscope.

    Activity Assay:

    Article Title: miR-423-5p-enriched small extracellular vesicles drive periodontal regeneration via Sfrp2+ cell expansion
    Article Snippet: For immunofluorescence staining, heart, liver, spleen, lung, kidney, and mandible samples were frozen in optimal cutting temperature compound (O.C.T. compound, Sakura Finetek, USA) and sectioned (5 μm).For immunofluorescence staining, heart, liver, spleen, lung, kidney, and mandible samples were frozen in optimal cutting temperature compound (O.C.T. compound, Sakura Finetek, USA) and sectioned (5 μm).. The slides were mounted with DAPI anti-fluorescence quenching mounting (Beyotime, Shanghai, China).. Observations and image collection were performed using a confocal microscope.Observations and image collection were performed using a confocal microscope.

    Western Blot:

    Article Title: miR-423-5p-enriched small extracellular vesicles drive periodontal regeneration via Sfrp2+ cell expansion
    Article Snippet: For immunofluorescence staining, heart, liver, spleen, lung, kidney, and mandible samples were frozen in optimal cutting temperature compound (O.C.T. compound, Sakura Finetek, USA) and sectioned (5 μm).For immunofluorescence staining, heart, liver, spleen, lung, kidney, and mandible samples were frozen in optimal cutting temperature compound (O.C.T. compound, Sakura Finetek, USA) and sectioned (5 μm).. The slides were mounted with DAPI anti-fluorescence quenching mounting (Beyotime, Shanghai, China).. Observations and image collection were performed using a confocal microscope.Observations and image collection were performed using a confocal microscope.

    Expressing:

    Article Title: miR-423-5p-enriched small extracellular vesicles drive periodontal regeneration via Sfrp2+ cell expansion
    Article Snippet: For immunofluorescence staining, heart, liver, spleen, lung, kidney, and mandible samples were frozen in optimal cutting temperature compound (O.C.T. compound, Sakura Finetek, USA) and sectioned (5 μm).For immunofluorescence staining, heart, liver, spleen, lung, kidney, and mandible samples were frozen in optimal cutting temperature compound (O.C.T. compound, Sakura Finetek, USA) and sectioned (5 μm).. The slides were mounted with DAPI anti-fluorescence quenching mounting (Beyotime, Shanghai, China).. Observations and image collection were performed using a confocal microscope.Observations and image collection were performed using a confocal microscope.

    Transfection:

    Article Title: miR-423-5p-enriched small extracellular vesicles drive periodontal regeneration via Sfrp2+ cell expansion
    Article Snippet: For immunofluorescence staining, heart, liver, spleen, lung, kidney, and mandible samples were frozen in optimal cutting temperature compound (O.C.T. compound, Sakura Finetek, USA) and sectioned (5 μm).For immunofluorescence staining, heart, liver, spleen, lung, kidney, and mandible samples were frozen in optimal cutting temperature compound (O.C.T. compound, Sakura Finetek, USA) and sectioned (5 μm).. The slides were mounted with DAPI anti-fluorescence quenching mounting (Beyotime, Shanghai, China).. Observations and image collection were performed using a confocal microscope.Observations and image collection were performed using a confocal microscope.

    Over Expression:

    Article Title: miR-423-5p-enriched small extracellular vesicles drive periodontal regeneration via Sfrp2+ cell expansion
    Article Snippet: For immunofluorescence staining, heart, liver, spleen, lung, kidney, and mandible samples were frozen in optimal cutting temperature compound (O.C.T. compound, Sakura Finetek, USA) and sectioned (5 μm).For immunofluorescence staining, heart, liver, spleen, lung, kidney, and mandible samples were frozen in optimal cutting temperature compound (O.C.T. compound, Sakura Finetek, USA) and sectioned (5 μm).. The slides were mounted with DAPI anti-fluorescence quenching mounting (Beyotime, Shanghai, China).. Observations and image collection were performed using a confocal microscope.Observations and image collection were performed using a confocal microscope.

    Plasmid Preparation:

    Article Title: miR-423-5p-enriched small extracellular vesicles drive periodontal regeneration via Sfrp2+ cell expansion
    Article Snippet: For immunofluorescence staining, heart, liver, spleen, lung, kidney, and mandible samples were frozen in optimal cutting temperature compound (O.C.T. compound, Sakura Finetek, USA) and sectioned (5 μm).For immunofluorescence staining, heart, liver, spleen, lung, kidney, and mandible samples were frozen in optimal cutting temperature compound (O.C.T. compound, Sakura Finetek, USA) and sectioned (5 μm).. The slides were mounted with DAPI anti-fluorescence quenching mounting (Beyotime, Shanghai, China).. Observations and image collection were performed using a confocal microscope.Observations and image collection were performed using a confocal microscope.

    Cytometry:

    Article Title: miR-423-5p-enriched small extracellular vesicles drive periodontal regeneration via Sfrp2+ cell expansion
    Article Snippet: For immunofluorescence staining, heart, liver, spleen, lung, kidney, and mandible samples were frozen in optimal cutting temperature compound (O.C.T. compound, Sakura Finetek, USA) and sectioned (5 μm).For immunofluorescence staining, heart, liver, spleen, lung, kidney, and mandible samples were frozen in optimal cutting temperature compound (O.C.T. compound, Sakura Finetek, USA) and sectioned (5 μm).. The slides were mounted with DAPI anti-fluorescence quenching mounting (Beyotime, Shanghai, China).. Observations and image collection were performed using a confocal microscope.Observations and image collection were performed using a confocal microscope.

    Zeta Potential Analyzer:

    Article Title: miR-423-5p-enriched small extracellular vesicles drive periodontal regeneration via Sfrp2+ cell expansion
    Article Snippet: For immunofluorescence staining, heart, liver, spleen, lung, kidney, and mandible samples were frozen in optimal cutting temperature compound (O.C.T. compound, Sakura Finetek, USA) and sectioned (5 μm).For immunofluorescence staining, heart, liver, spleen, lung, kidney, and mandible samples were frozen in optimal cutting temperature compound (O.C.T. compound, Sakura Finetek, USA) and sectioned (5 μm).. The slides were mounted with DAPI anti-fluorescence quenching mounting (Beyotime, Shanghai, China).. Observations and image collection were performed using a confocal microscope.Observations and image collection were performed using a confocal microscope.

    Real-time Polymerase Chain Reaction:

    Article Title: miR-423-5p-enriched small extracellular vesicles drive periodontal regeneration via Sfrp2+ cell expansion
    Article Snippet: For immunofluorescence staining, heart, liver, spleen, lung, kidney, and mandible samples were frozen in optimal cutting temperature compound (O.C.T. compound, Sakura Finetek, USA) and sectioned (5 μm).For immunofluorescence staining, heart, liver, spleen, lung, kidney, and mandible samples were frozen in optimal cutting temperature compound (O.C.T. compound, Sakura Finetek, USA) and sectioned (5 μm).. The slides were mounted with DAPI anti-fluorescence quenching mounting (Beyotime, Shanghai, China).. Observations and image collection were performed using a confocal microscope.Observations and image collection were performed using a confocal microscope.

    Confocal Microscopy:

    Article Title: miR-423-5p-enriched small extracellular vesicles drive periodontal regeneration via Sfrp2+ cell expansion
    Article Snippet: For immunofluorescence staining, heart, liver, spleen, lung, kidney, and mandible samples were frozen in optimal cutting temperature compound (O.C.T. compound, Sakura Finetek, USA) and sectioned (5 μm).For immunofluorescence staining, heart, liver, spleen, lung, kidney, and mandible samples were frozen in optimal cutting temperature compound (O.C.T. compound, Sakura Finetek, USA) and sectioned (5 μm).. The slides were mounted with DAPI anti-fluorescence quenching mounting (Beyotime, Shanghai, China).. Observations and image collection were performed using a confocal microscope.Observations and image collection were performed using a confocal microscope.

    Imaging:

    Article Title: miR-423-5p-enriched small extracellular vesicles drive periodontal regeneration via Sfrp2+ cell expansion
    Article Snippet: For immunofluorescence staining, heart, liver, spleen, lung, kidney, and mandible samples were frozen in optimal cutting temperature compound (O.C.T. compound, Sakura Finetek, USA) and sectioned (5 μm).For immunofluorescence staining, heart, liver, spleen, lung, kidney, and mandible samples were frozen in optimal cutting temperature compound (O.C.T. compound, Sakura Finetek, USA) and sectioned (5 μm).. The slides were mounted with DAPI anti-fluorescence quenching mounting (Beyotime, Shanghai, China).. Observations and image collection were performed using a confocal microscope.Observations and image collection were performed using a confocal microscope.

    Gene Expression:

    Article Title: miR-423-5p-enriched small extracellular vesicles drive periodontal regeneration via Sfrp2+ cell expansion
    Article Snippet: For immunofluorescence staining, heart, liver, spleen, lung, kidney, and mandible samples were frozen in optimal cutting temperature compound (O.C.T. compound, Sakura Finetek, USA) and sectioned (5 μm).For immunofluorescence staining, heart, liver, spleen, lung, kidney, and mandible samples were frozen in optimal cutting temperature compound (O.C.T. compound, Sakura Finetek, USA) and sectioned (5 μm).. The slides were mounted with DAPI anti-fluorescence quenching mounting (Beyotime, Shanghai, China).. Observations and image collection were performed using a confocal microscope.Observations and image collection were performed using a confocal microscope.



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    miR-423–5p was identified as a key osteogenic factor in sEVs. (A) Schematic illustration of the experimental design used to assess the impact of miR-423–5p on PDLC proliferation and osteogenic differentiation. n = 3. ( B ) EdU assay evaluating the influence of miR-423–5p on PDLC proliferation. Green fluorescence indicates EdU-labeled cells, and blue fluorescence indicates <t>DAPI-stained</t> nuclei. Scale bar = 100 μm. ( C ) Quantitative assessment of EdU-positive cell proportions. n = 5. ( D ) CCK-8 experiment evaluating the impact of miR-423–5p on PDLC proliferation. n = 6. ( E ) ALP and ARS assays assessing the impact of miR-423–5p on PDLC osteogenic differentiation. Scale bar = 200 μm. ( F ) Quantitative analysis of ALP activity. n = 6. ( G ) Western blot analysis of the influence of miR-423–5p on the osteogenic proteins expression (COL1, Runx2) in PDLCs. ( H ) Quantitative analysis of protein expression. n = 3. ( I ) The influence of miR-423–5p on the expression of osteogenic genes COL1 and Runx2 in PDLCs, evaluated using a PCR assay. n = 3. ( J ) Venn diagram showing the target genes predicted by miRTarBase, miRWalk, and TargetScan. ( K ) Pathway interaction map of the intersecting target genes. ( L ) ALP and ARS assays assessing the impact of PLCB1 on osteogenic differentiation in PDLCs. Scale bar = 200 μm. ( M ) Quantitative analysis of ALP activity. n = 6. ( N ) ALP staining and quantitative analysis of ALP activity in PDLCs co-transfected with miR-423–5p mimic and PLCB1 overexpression plasmid (n = 6). ( O ) Effects of miR-423–5p mimic combined with PLCB1 overexpression on the expression levels of PLCB1, Wnt3, and β-catenin in PDLCs.
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    miR-423–5p was identified as a key osteogenic factor in sEVs. (A) Schematic illustration of the experimental design used to assess the impact of miR-423–5p on PDLC proliferation and osteogenic differentiation. n = 3. ( B ) EdU assay evaluating the influence of miR-423–5p on PDLC proliferation. Green fluorescence indicates EdU-labeled cells, and blue fluorescence indicates <t>DAPI-stained</t> nuclei. Scale bar = 100 μm. ( C ) Quantitative assessment of EdU-positive cell proportions. n = 5. ( D ) CCK-8 experiment evaluating the impact of miR-423–5p on PDLC proliferation. n = 6. ( E ) ALP and ARS assays assessing the impact of miR-423–5p on PDLC osteogenic differentiation. Scale bar = 200 μm. ( F ) Quantitative analysis of ALP activity. n = 6. ( G ) Western blot analysis of the influence of miR-423–5p on the osteogenic proteins expression (COL1, Runx2) in PDLCs. ( H ) Quantitative analysis of protein expression. n = 3. ( I ) The influence of miR-423–5p on the expression of osteogenic genes COL1 and Runx2 in PDLCs, evaluated using a PCR assay. n = 3. ( J ) Venn diagram showing the target genes predicted by miRTarBase, miRWalk, and TargetScan. ( K ) Pathway interaction map of the intersecting target genes. ( L ) ALP and ARS assays assessing the impact of PLCB1 on osteogenic differentiation in PDLCs. Scale bar = 200 μm. ( M ) Quantitative analysis of ALP activity. n = 6. ( N ) ALP staining and quantitative analysis of ALP activity in PDLCs co-transfected with miR-423–5p mimic and PLCB1 overexpression plasmid (n = 6). ( O ) Effects of miR-423–5p mimic combined with PLCB1 overexpression on the expression levels of PLCB1, Wnt3, and β-catenin in PDLCs.
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    miR-423–5p was identified as a key osteogenic factor in sEVs. (A) Schematic illustration of the experimental design used to assess the impact of miR-423–5p on PDLC proliferation and osteogenic differentiation. n = 3. ( B ) EdU assay evaluating the influence of miR-423–5p on PDLC proliferation. Green fluorescence indicates EdU-labeled cells, and blue fluorescence indicates <t>DAPI-stained</t> nuclei. Scale bar = 100 μm. ( C ) Quantitative assessment of EdU-positive cell proportions. n = 5. ( D ) CCK-8 experiment evaluating the impact of miR-423–5p on PDLC proliferation. n = 6. ( E ) ALP and ARS assays assessing the impact of miR-423–5p on PDLC osteogenic differentiation. Scale bar = 200 μm. ( F ) Quantitative analysis of ALP activity. n = 6. ( G ) Western blot analysis of the influence of miR-423–5p on the osteogenic proteins expression (COL1, Runx2) in PDLCs. ( H ) Quantitative analysis of protein expression. n = 3. ( I ) The influence of miR-423–5p on the expression of osteogenic genes COL1 and Runx2 in PDLCs, evaluated using a PCR assay. n = 3. ( J ) Venn diagram showing the target genes predicted by miRTarBase, miRWalk, and TargetScan. ( K ) Pathway interaction map of the intersecting target genes. ( L ) ALP and ARS assays assessing the impact of PLCB1 on osteogenic differentiation in PDLCs. Scale bar = 200 μm. ( M ) Quantitative analysis of ALP activity. n = 6. ( N ) ALP staining and quantitative analysis of ALP activity in PDLCs co-transfected with miR-423–5p mimic and PLCB1 overexpression plasmid (n = 6). ( O ) Effects of miR-423–5p mimic combined with PLCB1 overexpression on the expression levels of PLCB1, Wnt3, and β-catenin in PDLCs.
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    miR-423–5p was identified as a key osteogenic factor in sEVs. (A) Schematic illustration of the experimental design used to assess the impact of miR-423–5p on PDLC proliferation and osteogenic differentiation. n = 3. ( B ) EdU assay evaluating the influence of miR-423–5p on PDLC proliferation. Green fluorescence indicates EdU-labeled cells, and blue fluorescence indicates <t>DAPI-stained</t> nuclei. Scale bar = 100 μm. ( C ) Quantitative assessment of EdU-positive cell proportions. n = 5. ( D ) CCK-8 experiment evaluating the impact of miR-423–5p on PDLC proliferation. n = 6. ( E ) ALP and ARS assays assessing the impact of miR-423–5p on PDLC osteogenic differentiation. Scale bar = 200 μm. ( F ) Quantitative analysis of ALP activity. n = 6. ( G ) Western blot analysis of the influence of miR-423–5p on the osteogenic proteins expression (COL1, Runx2) in PDLCs. ( H ) Quantitative analysis of protein expression. n = 3. ( I ) The influence of miR-423–5p on the expression of osteogenic genes COL1 and Runx2 in PDLCs, evaluated using a PCR assay. n = 3. ( J ) Venn diagram showing the target genes predicted by miRTarBase, miRWalk, and TargetScan. ( K ) Pathway interaction map of the intersecting target genes. ( L ) ALP and ARS assays assessing the impact of PLCB1 on osteogenic differentiation in PDLCs. Scale bar = 200 μm. ( M ) Quantitative analysis of ALP activity. n = 6. ( N ) ALP staining and quantitative analysis of ALP activity in PDLCs co-transfected with miR-423–5p mimic and PLCB1 overexpression plasmid (n = 6). ( O ) Effects of miR-423–5p mimic combined with PLCB1 overexpression on the expression levels of PLCB1, Wnt3, and β-catenin in PDLCs.
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    miR-423–5p was identified as a key osteogenic factor in sEVs. (A) Schematic illustration of the experimental design used to assess the impact of miR-423–5p on PDLC proliferation and osteogenic differentiation. n = 3. ( B ) EdU assay evaluating the influence of miR-423–5p on PDLC proliferation. Green fluorescence indicates EdU-labeled cells, and blue fluorescence indicates <t>DAPI-stained</t> nuclei. Scale bar = 100 μm. ( C ) Quantitative assessment of EdU-positive cell proportions. n = 5. ( D ) CCK-8 experiment evaluating the impact of miR-423–5p on PDLC proliferation. n = 6. ( E ) ALP and ARS assays assessing the impact of miR-423–5p on PDLC osteogenic differentiation. Scale bar = 200 μm. ( F ) Quantitative analysis of ALP activity. n = 6. ( G ) Western blot analysis of the influence of miR-423–5p on the osteogenic proteins expression (COL1, Runx2) in PDLCs. ( H ) Quantitative analysis of protein expression. n = 3. ( I ) The influence of miR-423–5p on the expression of osteogenic genes COL1 and Runx2 in PDLCs, evaluated using a PCR assay. n = 3. ( J ) Venn diagram showing the target genes predicted by miRTarBase, miRWalk, and TargetScan. ( K ) Pathway interaction map of the intersecting target genes. ( L ) ALP and ARS assays assessing the impact of PLCB1 on osteogenic differentiation in PDLCs. Scale bar = 200 μm. ( M ) Quantitative analysis of ALP activity. n = 6. ( N ) ALP staining and quantitative analysis of ALP activity in PDLCs co-transfected with miR-423–5p mimic and PLCB1 overexpression plasmid (n = 6). ( O ) Effects of miR-423–5p mimic combined with PLCB1 overexpression on the expression levels of PLCB1, Wnt3, and β-catenin in PDLCs.
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    miR-423–5p was identified as a key osteogenic factor in sEVs. (A) Schematic illustration of the experimental design used to assess the impact of miR-423–5p on PDLC proliferation and osteogenic differentiation. n = 3. ( B ) EdU assay evaluating the influence of miR-423–5p on PDLC proliferation. Green fluorescence indicates EdU-labeled cells, and blue fluorescence indicates <t>DAPI-stained</t> nuclei. Scale bar = 100 μm. ( C ) Quantitative assessment of EdU-positive cell proportions. n = 5. ( D ) CCK-8 experiment evaluating the impact of miR-423–5p on PDLC proliferation. n = 6. ( E ) ALP and ARS assays assessing the impact of miR-423–5p on PDLC osteogenic differentiation. Scale bar = 200 μm. ( F ) Quantitative analysis of ALP activity. n = 6. ( G ) Western blot analysis of the influence of miR-423–5p on the osteogenic proteins expression (COL1, Runx2) in PDLCs. ( H ) Quantitative analysis of protein expression. n = 3. ( I ) The influence of miR-423–5p on the expression of osteogenic genes COL1 and Runx2 in PDLCs, evaluated using a PCR assay. n = 3. ( J ) Venn diagram showing the target genes predicted by miRTarBase, miRWalk, and TargetScan. ( K ) Pathway interaction map of the intersecting target genes. ( L ) ALP and ARS assays assessing the impact of PLCB1 on osteogenic differentiation in PDLCs. Scale bar = 200 μm. ( M ) Quantitative analysis of ALP activity. n = 6. ( N ) ALP staining and quantitative analysis of ALP activity in PDLCs co-transfected with miR-423–5p mimic and PLCB1 overexpression plasmid (n = 6). ( O ) Effects of miR-423–5p mimic combined with PLCB1 overexpression on the expression levels of PLCB1, Wnt3, and β-catenin in PDLCs.
    Anti Fluorescence Quenching Mounting Medium, supplied by Beyotime, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    miR-423–5p was identified as a key osteogenic factor in sEVs. (A) Schematic illustration of the experimental design used to assess the impact of miR-423–5p on PDLC proliferation and osteogenic differentiation. n = 3. ( B ) EdU assay evaluating the influence of miR-423–5p on PDLC proliferation. Green fluorescence indicates EdU-labeled cells, and blue fluorescence indicates <t>DAPI-stained</t> nuclei. Scale bar = 100 μm. ( C ) Quantitative assessment of EdU-positive cell proportions. n = 5. ( D ) CCK-8 experiment evaluating the impact of miR-423–5p on PDLC proliferation. n = 6. ( E ) ALP and ARS assays assessing the impact of miR-423–5p on PDLC osteogenic differentiation. Scale bar = 200 μm. ( F ) Quantitative analysis of ALP activity. n = 6. ( G ) Western blot analysis of the influence of miR-423–5p on the osteogenic proteins expression (COL1, Runx2) in PDLCs. ( H ) Quantitative analysis of protein expression. n = 3. ( I ) The influence of miR-423–5p on the expression of osteogenic genes COL1 and Runx2 in PDLCs, evaluated using a PCR assay. n = 3. ( J ) Venn diagram showing the target genes predicted by miRTarBase, miRWalk, and TargetScan. ( K ) Pathway interaction map of the intersecting target genes. ( L ) ALP and ARS assays assessing the impact of PLCB1 on osteogenic differentiation in PDLCs. Scale bar = 200 μm. ( M ) Quantitative analysis of ALP activity. n = 6. ( N ) ALP staining and quantitative analysis of ALP activity in PDLCs co-transfected with miR-423–5p mimic and PLCB1 overexpression plasmid (n = 6). ( O ) Effects of miR-423–5p mimic combined with PLCB1 overexpression on the expression levels of PLCB1, Wnt3, and β-catenin in PDLCs.
    Dapi Anti Fluorescence Quenching Mounting Medium, supplied by Beyotime, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    miR-423–5p was identified as a key osteogenic factor in sEVs. (A) Schematic illustration of the experimental design used to assess the impact of miR-423–5p on PDLC proliferation and osteogenic differentiation. n = 3. ( B ) EdU assay evaluating the influence of miR-423–5p on PDLC proliferation. Green fluorescence indicates EdU-labeled cells, and blue fluorescence indicates <t>DAPI-stained</t> nuclei. Scale bar = 100 μm. ( C ) Quantitative assessment of EdU-positive cell proportions. n = 5. ( D ) CCK-8 experiment evaluating the impact of miR-423–5p on PDLC proliferation. n = 6. ( E ) ALP and ARS assays assessing the impact of miR-423–5p on PDLC osteogenic differentiation. Scale bar = 200 μm. ( F ) Quantitative analysis of ALP activity. n = 6. ( G ) Western blot analysis of the influence of miR-423–5p on the osteogenic proteins expression (COL1, Runx2) in PDLCs. ( H ) Quantitative analysis of protein expression. n = 3. ( I ) The influence of miR-423–5p on the expression of osteogenic genes COL1 and Runx2 in PDLCs, evaluated using a PCR assay. n = 3. ( J ) Venn diagram showing the target genes predicted by miRTarBase, miRWalk, and TargetScan. ( K ) Pathway interaction map of the intersecting target genes. ( L ) ALP and ARS assays assessing the impact of PLCB1 on osteogenic differentiation in PDLCs. Scale bar = 200 μm. ( M ) Quantitative analysis of ALP activity. n = 6. ( N ) ALP staining and quantitative analysis of ALP activity in PDLCs co-transfected with miR-423–5p mimic and PLCB1 overexpression plasmid (n = 6). ( O ) Effects of miR-423–5p mimic combined with PLCB1 overexpression on the expression levels of PLCB1, Wnt3, and β-catenin in PDLCs.
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    miR-423–5p was identified as a key osteogenic factor in sEVs. (A) Schematic illustration of the experimental design used to assess the impact of miR-423–5p on PDLC proliferation and osteogenic differentiation. n = 3. ( B ) EdU assay evaluating the influence of miR-423–5p on PDLC proliferation. Green fluorescence indicates EdU-labeled cells, and blue fluorescence indicates <t>DAPI-stained</t> nuclei. Scale bar = 100 μm. ( C ) Quantitative assessment of EdU-positive cell proportions. n = 5. ( D ) CCK-8 experiment evaluating the impact of miR-423–5p on PDLC proliferation. n = 6. ( E ) ALP and ARS assays assessing the impact of miR-423–5p on PDLC osteogenic differentiation. Scale bar = 200 μm. ( F ) Quantitative analysis of ALP activity. n = 6. ( G ) Western blot analysis of the influence of miR-423–5p on the osteogenic proteins expression (COL1, Runx2) in PDLCs. ( H ) Quantitative analysis of protein expression. n = 3. ( I ) The influence of miR-423–5p on the expression of osteogenic genes COL1 and Runx2 in PDLCs, evaluated using a PCR assay. n = 3. ( J ) Venn diagram showing the target genes predicted by miRTarBase, miRWalk, and TargetScan. ( K ) Pathway interaction map of the intersecting target genes. ( L ) ALP and ARS assays assessing the impact of PLCB1 on osteogenic differentiation in PDLCs. Scale bar = 200 μm. ( M ) Quantitative analysis of ALP activity. n = 6. ( N ) ALP staining and quantitative analysis of ALP activity in PDLCs co-transfected with miR-423–5p mimic and PLCB1 overexpression plasmid (n = 6). ( O ) Effects of miR-423–5p mimic combined with PLCB1 overexpression on the expression levels of PLCB1, Wnt3, and β-catenin in PDLCs.
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    miR-423–5p was identified as a key osteogenic factor in sEVs. (A) Schematic illustration of the experimental design used to assess the impact of miR-423–5p on PDLC proliferation and osteogenic differentiation. n = 3. ( B ) EdU assay evaluating the influence of miR-423–5p on PDLC proliferation. Green fluorescence indicates EdU-labeled cells, and blue fluorescence indicates DAPI-stained nuclei. Scale bar = 100 μm. ( C ) Quantitative assessment of EdU-positive cell proportions. n = 5. ( D ) CCK-8 experiment evaluating the impact of miR-423–5p on PDLC proliferation. n = 6. ( E ) ALP and ARS assays assessing the impact of miR-423–5p on PDLC osteogenic differentiation. Scale bar = 200 μm. ( F ) Quantitative analysis of ALP activity. n = 6. ( G ) Western blot analysis of the influence of miR-423–5p on the osteogenic proteins expression (COL1, Runx2) in PDLCs. ( H ) Quantitative analysis of protein expression. n = 3. ( I ) The influence of miR-423–5p on the expression of osteogenic genes COL1 and Runx2 in PDLCs, evaluated using a PCR assay. n = 3. ( J ) Venn diagram showing the target genes predicted by miRTarBase, miRWalk, and TargetScan. ( K ) Pathway interaction map of the intersecting target genes. ( L ) ALP and ARS assays assessing the impact of PLCB1 on osteogenic differentiation in PDLCs. Scale bar = 200 μm. ( M ) Quantitative analysis of ALP activity. n = 6. ( N ) ALP staining and quantitative analysis of ALP activity in PDLCs co-transfected with miR-423–5p mimic and PLCB1 overexpression plasmid (n = 6). ( O ) Effects of miR-423–5p mimic combined with PLCB1 overexpression on the expression levels of PLCB1, Wnt3, and β-catenin in PDLCs.

    Journal: Bioactive Materials

    Article Title: miR-423-5p-enriched small extracellular vesicles drive periodontal regeneration via Sfrp2+ cell expansion

    doi: 10.1016/j.bioactmat.2025.11.026

    Figure Lengend Snippet: miR-423–5p was identified as a key osteogenic factor in sEVs. (A) Schematic illustration of the experimental design used to assess the impact of miR-423–5p on PDLC proliferation and osteogenic differentiation. n = 3. ( B ) EdU assay evaluating the influence of miR-423–5p on PDLC proliferation. Green fluorescence indicates EdU-labeled cells, and blue fluorescence indicates DAPI-stained nuclei. Scale bar = 100 μm. ( C ) Quantitative assessment of EdU-positive cell proportions. n = 5. ( D ) CCK-8 experiment evaluating the impact of miR-423–5p on PDLC proliferation. n = 6. ( E ) ALP and ARS assays assessing the impact of miR-423–5p on PDLC osteogenic differentiation. Scale bar = 200 μm. ( F ) Quantitative analysis of ALP activity. n = 6. ( G ) Western blot analysis of the influence of miR-423–5p on the osteogenic proteins expression (COL1, Runx2) in PDLCs. ( H ) Quantitative analysis of protein expression. n = 3. ( I ) The influence of miR-423–5p on the expression of osteogenic genes COL1 and Runx2 in PDLCs, evaluated using a PCR assay. n = 3. ( J ) Venn diagram showing the target genes predicted by miRTarBase, miRWalk, and TargetScan. ( K ) Pathway interaction map of the intersecting target genes. ( L ) ALP and ARS assays assessing the impact of PLCB1 on osteogenic differentiation in PDLCs. Scale bar = 200 μm. ( M ) Quantitative analysis of ALP activity. n = 6. ( N ) ALP staining and quantitative analysis of ALP activity in PDLCs co-transfected with miR-423–5p mimic and PLCB1 overexpression plasmid (n = 6). ( O ) Effects of miR-423–5p mimic combined with PLCB1 overexpression on the expression levels of PLCB1, Wnt3, and β-catenin in PDLCs.

    Article Snippet: The slides were mounted with DAPI anti-fluorescence quenching mounting (Beyotime, Shanghai, China).

    Techniques: EdU Assay, Fluorescence, Labeling, Staining, CCK-8 Assay, Activity Assay, Western Blot, Expressing, Transfection, Over Expression, Plasmid Preparation

    sEVs miR−423−5p increased the osteogenic differentiation potential of sEVs. (A) TEM characterization of the morphology of sEVs and sEVs miR−423−5p morphology. Scale bar = 200 nm. ( B ) Nanoflow cytometry of the particle size distribution of sEVs and sEVs miR−423−5p . ( C ) Western blot analysis of sEVs and sEVs miR−423−5p markers, including CD9, CD63, HSP90, TSG101, and the endoplasmic reticulum protein calnexin. ( D ) Analysis of the Zeta potential of sEVs and sEVs miR−423−5p . n = 3. ( E ) Quantitative PCR analysis comparing miR-423–5p expression levels between sEVs and sEVs miR−423−5p . n = 3. ( F ) Confocal microscopy imaging of sEV/sEVs miR−423−5p uptake by PDLCs. PDLC nuclei are marked with DAPI (blue), sEVs stained with PKH26 (red dots), the cytoskeleton stained with Cell Mask Actin (red filaments), and miR-423–5p is depicted in green. Scale bar = 20 μm. ( G ) Quantitative PCR assessment of miR-423–5p, miR-100–5p, miR-125b-5p, miR-26a-5p, and miR-24–3p levels in PDLCs following sEV/sEVs miR−423−5p uptake. n = 3. ( H ) RIP-qPCR analysis showing that miR-423–5p delivered by sEVs miR−423−5p is loaded onto Ago2 (Argonaute 2) protein in PDLCs. n = 3. ( I ) CCK-8 experiment evaluating the impact of sEVs miR−423−5p on PDLC proliferation. n = 4. ( J ) Western blot analysis of proliferative markers Ki67, MCM7, and PCNA (proliferating cell nuclear antigen) in sEVs miR−423−5p -treated PDLCs. ( K ) Quantitative analysis of protein expression. n = 3. ( L ) ALP and ARS assays assessing the influence of sEVs miR−423−5p on mediating PDLC osteogenic differentiation. Scale bar = 200 μm. ( M ) Quantitative analysis of ALP activity. n = 6. ( N ) PCR analysis of osteogenic gene expression in PDLCs following sEVs miR−423−5p treatment. n = 3. ( O ) Western blot quantification of osteogenic protein levels in sEVs miR−423−5p -treated PDLCs. ( P ) Quantitative analysis of protein expression. n = 3.

    Journal: Bioactive Materials

    Article Title: miR-423-5p-enriched small extracellular vesicles drive periodontal regeneration via Sfrp2+ cell expansion

    doi: 10.1016/j.bioactmat.2025.11.026

    Figure Lengend Snippet: sEVs miR−423−5p increased the osteogenic differentiation potential of sEVs. (A) TEM characterization of the morphology of sEVs and sEVs miR−423−5p morphology. Scale bar = 200 nm. ( B ) Nanoflow cytometry of the particle size distribution of sEVs and sEVs miR−423−5p . ( C ) Western blot analysis of sEVs and sEVs miR−423−5p markers, including CD9, CD63, HSP90, TSG101, and the endoplasmic reticulum protein calnexin. ( D ) Analysis of the Zeta potential of sEVs and sEVs miR−423−5p . n = 3. ( E ) Quantitative PCR analysis comparing miR-423–5p expression levels between sEVs and sEVs miR−423−5p . n = 3. ( F ) Confocal microscopy imaging of sEV/sEVs miR−423−5p uptake by PDLCs. PDLC nuclei are marked with DAPI (blue), sEVs stained with PKH26 (red dots), the cytoskeleton stained with Cell Mask Actin (red filaments), and miR-423–5p is depicted in green. Scale bar = 20 μm. ( G ) Quantitative PCR assessment of miR-423–5p, miR-100–5p, miR-125b-5p, miR-26a-5p, and miR-24–3p levels in PDLCs following sEV/sEVs miR−423−5p uptake. n = 3. ( H ) RIP-qPCR analysis showing that miR-423–5p delivered by sEVs miR−423−5p is loaded onto Ago2 (Argonaute 2) protein in PDLCs. n = 3. ( I ) CCK-8 experiment evaluating the impact of sEVs miR−423−5p on PDLC proliferation. n = 4. ( J ) Western blot analysis of proliferative markers Ki67, MCM7, and PCNA (proliferating cell nuclear antigen) in sEVs miR−423−5p -treated PDLCs. ( K ) Quantitative analysis of protein expression. n = 3. ( L ) ALP and ARS assays assessing the influence of sEVs miR−423−5p on mediating PDLC osteogenic differentiation. Scale bar = 200 μm. ( M ) Quantitative analysis of ALP activity. n = 6. ( N ) PCR analysis of osteogenic gene expression in PDLCs following sEVs miR−423−5p treatment. n = 3. ( O ) Western blot quantification of osteogenic protein levels in sEVs miR−423−5p -treated PDLCs. ( P ) Quantitative analysis of protein expression. n = 3.

    Article Snippet: The slides were mounted with DAPI anti-fluorescence quenching mounting (Beyotime, Shanghai, China).

    Techniques: Cytometry, Western Blot, Zeta Potential Analyzer, Real-time Polymerase Chain Reaction, Expressing, Confocal Microscopy, Imaging, Staining, CCK-8 Assay, Activity Assay, Gene Expression

    sEVs miR−423−5p promoted periodontal regeneration in rats. ( A ) Morphological characterization of collagen sponges and sEV/sEVs miR−423−5p -loaded collagen sponges via SEM. White arrows indicate sEVs and sEVs miR−423−5p . Scale bars: 2 μm, and 100 nm at increasing magnifications. ( B ) Analysis of the release rate of sEVs/sEVs miR−423−5p from collagen sponges. ( C ) Schematic representation of the periodontal defect model. ( D ) Confocal imaging revealed the distribution of sEVs and sEVs miR−423−5p within the periodontal defects in rats. PKH26-labeled sEVs are shown in red (red arrows), miR-423–5p in green (green arrows), colocalization of sEVs and miR-423–5p in orange (orange arrows), and DAPI-stained nuclei in blue. Scale bars: 500 μm, 100 μm, 50 μm, and 20 μm at increasing magnification. ( E ) CT images of periodontal defect regions in rats. Scale bar = 1 mm. ( F ) Quantitative analysis of new bone volume fraction (bone volume/tissue volume; BV/TV), bone mineral density (BMD), trabecular thickness (Tb. Th), and trabecular separation (Tb. Sp) in the defect areas. n = 4. ( G ) HE and Masson's trichrome staining of regenerated periodontal tissue, with black dashed lines indicating defect regions and red asterisks highlighting periodontal ligament-like structures. Scale bars: 625 μm (low magnification), 100 μm (high magnification). AB: alveolar bone; CS: collagen sponges; PDL: periodontal ligament; d: dentin.

    Journal: Bioactive Materials

    Article Title: miR-423-5p-enriched small extracellular vesicles drive periodontal regeneration via Sfrp2+ cell expansion

    doi: 10.1016/j.bioactmat.2025.11.026

    Figure Lengend Snippet: sEVs miR−423−5p promoted periodontal regeneration in rats. ( A ) Morphological characterization of collagen sponges and sEV/sEVs miR−423−5p -loaded collagen sponges via SEM. White arrows indicate sEVs and sEVs miR−423−5p . Scale bars: 2 μm, and 100 nm at increasing magnifications. ( B ) Analysis of the release rate of sEVs/sEVs miR−423−5p from collagen sponges. ( C ) Schematic representation of the periodontal defect model. ( D ) Confocal imaging revealed the distribution of sEVs and sEVs miR−423−5p within the periodontal defects in rats. PKH26-labeled sEVs are shown in red (red arrows), miR-423–5p in green (green arrows), colocalization of sEVs and miR-423–5p in orange (orange arrows), and DAPI-stained nuclei in blue. Scale bars: 500 μm, 100 μm, 50 μm, and 20 μm at increasing magnification. ( E ) CT images of periodontal defect regions in rats. Scale bar = 1 mm. ( F ) Quantitative analysis of new bone volume fraction (bone volume/tissue volume; BV/TV), bone mineral density (BMD), trabecular thickness (Tb. Th), and trabecular separation (Tb. Sp) in the defect areas. n = 4. ( G ) HE and Masson's trichrome staining of regenerated periodontal tissue, with black dashed lines indicating defect regions and red asterisks highlighting periodontal ligament-like structures. Scale bars: 625 μm (low magnification), 100 μm (high magnification). AB: alveolar bone; CS: collagen sponges; PDL: periodontal ligament; d: dentin.

    Article Snippet: The slides were mounted with DAPI anti-fluorescence quenching mounting (Beyotime, Shanghai, China).

    Techniques: Imaging, Labeling, Staining