Journal: Frontiers in Cell and Developmental Biology

Article Title: Optimal Sca-1-based procedure for purifying mouse adipose-derived mesenchymal stem cells with enhanced proliferative and differentiation potential

doi: 10.3389/fcell.2025.1566670

Figure Lengend Snippet: Differentiation potential of mouse ADSCs obtained by three purification methods. (A) After osteogenic induction culture, the cells were stained with alizarin red S, and the histogram showed the area of calcium nodules for quantitative analysis ( n = 5). (B) Oil red O staining after adipogenic induction cultures and histogram showing lipid droplet regions for quantitative analysis ( n = 5). (C) After chondrogenic induction culture, alcian blue staining and histograms showing quantitative sorting of mucopolysaccharide regions ( n = 5). Data were analyzed by one-way ANOVA and expressed as mean ± SEM. ns, not significant; * P < 0.05, ** P < 0.01, *** P < 0.001. Scale bar = 100 µm.

Article Snippet: In accordance with the manual for the osteogenic induction differentiation kit (Cyagen, Guangzhou, China), ADSC-A, ADSC-M, and ADSC-AM at P3 were seeded in six-well plates coated with 0.1% gelatin at a density of 2 × 10 4 cells/cm 2 .

Techniques: Purification, Staining

Journal: Frontiers in Cell and Developmental Biology

Article Title: Plant and animal-derived fusion nanovesicles rescue inflammation-compromised osteogenic potential of periodontal ligament stem cells

doi: 10.3389/fcell.2025.1512238

Figure Lengend Snippet: Effects of SBNVs on LPS or TNF-α-induced inflammatory response and osteogenic differentiation. (A) The effects of SBNVs on proliferation of Pg-LPS-induced PDLSCs (n = 3). (B) Relative mRNA expression of IL-1β, IL-6 and TNF-α in Pg-LPS-induced PDLSCs after SBNVs treatments (n = 3). (C) Flow cytometric analysis of intracellular ROS levels in PDLSCs. NC, cells without Pg-LPS treatment. (D, E) Typical Western blot bands and quantitative analysis of relative RUNX2 protein levels in Pg-LPS-stimulated PDLSCs at day 7 after SBNVs treatments. (F) Relative mRNA expression of RUNX2 in Pg-LPS-stimulated PDLSCs at day 7 after SBNVs treatments (n = 3). (G) Relative mRNA expression of RUNX2 in TNF-α-stimulated PDLSCs at day 7 after SBNVs treatments (n = 3). (H, I) Typical Western blot bands and quantitative analysis of relative protein levels of ALP and RUNX2 in TNF-α-stimulated PDLSCs at day 7 after SBNVs treatments. (J, K) Alizarin Red (J) and ALP (K) staining shows mineralized nodules of TNF-α-stimulated PDLSCs at days 7 and 21 after SBNVs treatments, respectively. Scale bars, 200 µm. For Panels (A–C) , cells in the NC group were treated with medium alone. For Panels (D–K) , cells in the NC group were treated with osteogenic differentiation medium alone. *p < 0.05; ***p < 0.001.

Article Snippet: According to the instructions, the osteogenic induction differentiation kit (Cyagen, Cat #HUXXC-90021) was used to induce PDLSCs for osteogenic differentiation.

Techniques: Expressing, Western Blot, Staining

Journal: Frontiers in Cell and Developmental Biology

Article Title: Plant and animal-derived fusion nanovesicles rescue inflammation-compromised osteogenic potential of periodontal ligament stem cells

doi: 10.3389/fcell.2025.1512238

Figure Lengend Snippet: TNFR1-NVs neutralized TNF-α and promoted PDLSCs osteogenic differentiation. (A) Relative mRNA expression of IL-10 and IDO1 in NVs from different mesenchymal stem cells (n = 3). (B) Flow cytometric analysis of intracellular ROS levels in PDLSCs. The blue portion represents the NC group, which was treated with medium alone. The experimental groups, indicated in red, were treated with TNF-α, either with or without the therapeutic agent. (C) Schematic diagram of the preparation of TNFR1-NVs. (D) Confocal image showing colocalization of TNFR1-GFP and DID-labeled cell membranes; DAPI was used to label cell nuclei. Scale bar: 10 μm. (E–G) Representative TEM image, zeta potential, size distribution of TNFR1-NVs. Scale bar: 100 nm. (H) Western blot analysis of the expression of TNFR1-GFP and MSC markers on TNFR1-NVs. WCL, whole cell lysate. (I) ELISA assay to detect the ability of NVs to neutralize TNF-α (n = 3). (J) Relative mRNA levels of IL-1β, IL-6 and TNF-α in TNF-α-stimulated PDLSCs after different NVs treatment (n = 3). (K) Relative mRNA expression of ALP and RUNX2 in TNF-α-stimulated PDLSCs at day 7 after different NVs treatments (n = 3). (L–M) Typical Western blot bands and quantitative analysis of relative protein levels of ALP and RUNX2 in TNF-α-stimulated PDLSCs at day 7 after different NVs treatments. For Panelss (J–M) , cells in the NC group were treated with osteogenic differentiation medium alone. *p < 0.05; **p < 0.01; ***p < 0.001.

Article Snippet: According to the instructions, the osteogenic induction differentiation kit (Cyagen, Cat #HUXXC-90021) was used to induce PDLSCs for osteogenic differentiation.

Techniques: Expressing, Labeling, Zeta Potential Analyzer, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: Plant and animal-derived fusion nanovesicles rescue inflammation-compromised osteogenic potential of periodontal ligament stem cells

doi: 10.3389/fcell.2025.1512238

Figure Lengend Snippet: FVs alleviated the inflammatory response and osteogenic differentiation inhibition of PDLSCs in the inflammatory microenvironment. (A) Relative mRNA levels of pro-inflammatory cytokines IL-1β and TNF-α in PDLSCs after different treatments (n = 3). (B, C) Typical Western blot bands and quantitative analysis of relative protein levels of IL-1β and TNF-α in PDLSCs after different treatments (n = 3). (D) Relative mRNA levels of ALP and RUNX2 in PDLSCs at day 7 after different treatments (n = 3). (E, F) Typical Western blot bands and quantitative analysis of relative protein levels of ALP and RUNX2 in PDLSCs at day 7 after different treatments (n = 3). (G, H) ALP (G) and Alizarin Red (H) staining shows mineralized nodules of PDLSCs in the inflammatory microenvironment at days 7 and 21 after different treatments, respectively. Scale bars, 200 µm. In all cases, 10 μg/mL Pg-LPS together with 20 ng/mL TNF-α were utilized to simulate the inflammatory microenvironment in vitro. For Panels (A–C) , cells in the NC group were treated with medium alone. For Panels (D–H) , cells in the NC group were treated with osteogenic differentiation medium alone. *p < 0.05; **p < 0.01; ***p < 0.001.

Article Snippet: According to the instructions, the osteogenic induction differentiation kit (Cyagen, Cat #HUXXC-90021) was used to induce PDLSCs for osteogenic differentiation.

Techniques: Inhibition, Western Blot, Staining, In Vitro

Journal: Frontiers in Cell and Developmental Biology

Article Title: Plant and animal-derived fusion nanovesicles rescue inflammation-compromised osteogenic potential of periodontal ligament stem cells

doi: 10.3389/fcell.2025.1512238

Figure Lengend Snippet: miR-21-5p of fused nanovesicles promoted osteogenic differentiation of PDLSCs by inhibiting STAT3 in the inflammatory microenvironment. (A) Venn diagram of miRNAs included in FVs and SBNVs obtained by miRNA-seq. (B) Heatmap of the top ten most abundant miRNAs obtained from FVs through miRNA-seq. (C) Relative percentage of each miRNA in the total miRNA read counts obtained from FVs through miRNA-seq. (D) Relative expression levels of miR-21-5p in FVs and TNFR1-NVs (n = 3). (E) The expression level of miR-21-5p was suppressed in the osteogenic differentiation process of PDLSCs in the inflammatory microenvironment (n = 3). (F–H) Western blot analysis of the protein levels of ALP and RUNX2 in PDLSCs treated with miR-21-5p mimics or mimics control in the inflammatory microenvironment at day 7 (n = 3). (I) Venn diagram showing the overlap between miR-21-5p target genes in three public databases and JAK-STAT pathway genes. (J) Western blot analysis of the activation level of STAT3 in PDLSCs overexpressing miR-21-5p at day 7 (n = 3). The values represent the relative ratio of p-STAT3 to total STAT3 for each sample. In all cases, 10 μg/mL Pg-LPS together with 20 ng/mL TNF-α were utilized to simulate the inflammatory microenvironment in vitro . *p < 0.05; **p < 0.01.

Article Snippet: According to the instructions, the osteogenic induction differentiation kit (Cyagen, Cat #HUXXC-90021) was used to induce PDLSCs for osteogenic differentiation.

Techniques: Expressing, Western Blot, Control, Activation Assay, In Vitro