plcb1  (Sangon Biotech)

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: Schematic diagram. Preparation and functional mechanisms of sEVs miR−423−5p in promoting periodontal regeneration. miR-423–5p was incubated with Cell-Penetrating Peptide (CPP) for 15 min at room temperature to form a complex. The positively charged surface of CPP promoted their adsorption onto negatively charged sEVs, facilitating entry through electrostatic and hydrophobic interactions. Following CPP degradation, engineered sEVs enriched with miR-423–5p (known as sEVs miR−423−5p ) were successfully generated. Once internalized by PDLCs via endocytosis, membrane fusion, or ligand–receptor interactions, sEVs miR−423−5p promoted PDLC proliferation. The released miR-423–5p silenced PLCB1, thereby enhancing osteogenic differentiation and facilitating the formation of the periodontal complex—including cementum, periodontal ligament, and alveolar bone—by expanding Sfrp2 + fibroblasts in the defect region.

Article Snippet: PDLCs were transfected with siPLCB1 (TSINGKE BIOTECH, Beijing, China) and PLCB1 overexpression plasmids (OBIO Technology, Shanghai, China) according to the LipofectamineTM 3000 protocol (Invitrogen, Carlsbad, CA, USA).

Techniques: Functional Assay, Incubation, Adsorption, Generated, Membrane

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: PDLCs were transfected with siPLCB1 (TSINGKE BIOTECH, Beijing, China) and PLCB1 overexpression plasmids (OBIO Technology, Shanghai, China) according to the LipofectamineTM 3000 protocol (Invitrogen, Carlsbad, CA, USA).

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