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

Journal: Thoracic Cancer

Article Title: LncRNA AC100826.1 regulated PLCB1 to promote progression in non‐small cell lung cancer

doi: 10.1111/1759-7714.15323

Figure Lengend Snippet: Primer sequences for reverse transcription‐quantitative polymerase chain reaction.

Article Snippet: Small interfering RNA (SiRNA) specifically targeting Lnc1 and PLCB1 was purchased from Sangon Biotechnology.

Techniques:

Journal: Thoracic Cancer

Article Title: LncRNA AC100826.1 regulated PLCB1 to promote progression in non‐small cell lung cancer

doi: 10.1111/1759-7714.15323

Figure Lengend Snippet: Lnc1 promotes non‐small cell lung cancer (NSCLC) cell proliferation and metastasis mediated by PLCB1. (a) Identifying potential target genes of the downstream of Lnc1 (Green:LncTarget data; Red: RNAseq). (b–e) Using reverse transcription‐quantitative polymerase chain reaction (RT‐qPCR) to quantifiability detection the level of potential target genes (ADCY7, NGF, PLCB1 and TLN2) after knockdown or overexpression of Lnc1 in A549 cells and H1299 cells. (f) Quantifiability detection of the mRNA expression level of PLCB1 in 10 pairs of clinical samples. (g) The prognostic value of PLCB1 in NSCLC examined by Kaplan–Meier Plotter. (h) The expression level of PLCB1 in patients with different degrees of differentiation and metastasis. (i, j) Transwell and scratch assays proved that PLCB1 can promote tumor cell migration (ns, no statistical significance, * p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: Small interfering RNA (SiRNA) specifically targeting Lnc1 and PLCB1 was purchased from Sangon Biotechnology.

Techniques: Reverse Transcription, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Knockdown, Over Expression, Expressing, Migration

Journal: Thoracic Cancer

Article Title: LncRNA AC100826.1 regulated PLCB1 to promote progression in non‐small cell lung cancer

doi: 10.1111/1759-7714.15323

Figure Lengend Snippet: Lnc1 promotes non‐small cell lung cancer (NSCLC) cell proliferation and metastasis mediated by PLCB1. (a) Western blots were used to analyze the expression level of epithelial mesenchymal transition (EMT)‐related proteins in A549 cells and H1299 cells. (b, c) RNA pulldown and RNA immunoprecipitation (RIP) experiments proved the interaction between PLCB1 and Lnc1. (d) Western blots were used to analyze the protein expression level of PLCB1 after knockdown Lnc1 in A549 cell and H1299 cell. (e, f) Cell counting kit‐8 (CCK‐8) and transwell assays were used to evaluate the interaction PLCB1 and Lnc1 on A549 cell and H1299 cell.(ns, no statistical significance, * p < 0.05, ** p < 0.01).

Article Snippet: Small interfering RNA (SiRNA) specifically targeting Lnc1 and PLCB1 was purchased from Sangon Biotechnology.

Techniques: Western Blot, Expressing, RNA Immunoprecipitation, Knockdown, Cell Counting, CCK-8 Assay

Journal: Thoracic Cancer

Article Title: LncRNA AC100826.1 regulated PLCB1 to promote progression in non‐small cell lung cancer

doi: 10.1111/1759-7714.15323

Figure Lengend Snippet: Lnc1 activates Rap1 signaling pathway by targeting PLCB1. (a) Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis was used to detect the signal regulation in shControl‐A549 group and shLnc1#1‐A549 group. (b) Western blots were used to analyze the expression level of Rap1 signal pathway marker protein in A549 cells and H1299 cells. (c) Transwell assay was used to measure the migration of A549 cells and H1229 cells after treatment with the Rap1 signaling pathway inhibitor (ERK‐IN3). (d) Western blots were used to evaluate the effect of the Rap1 signaling pathway inhibitor (ERK‐IN3) on epithelial mesenchymal transition (EMT) related proteins. (ns, no statistical significance, * p < 0.05, ** p < 0.01).

Article Snippet: Small interfering RNA (SiRNA) specifically targeting Lnc1 and PLCB1 was purchased from Sangon Biotechnology.

Techniques: Western Blot, Expressing, Marker, Transwell Assay, Migration