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

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

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