Journal: Biology

Article Title: Force-Dependent Presence of Senescent Cells Expressing Vascular Endothelial Growth Factor During Orthodontic Tooth Movement

doi: 10.3390/biology15020187

Figure Lengend Snippet: Dynamic changes in angiogenesis induced by OTM. ( A ) Representative CD31 immunofluorescence staining images. CD31 staining without DAPI ( left ) and merged images with DAPI ( right ) are shown. ( B ) Quantitative analysis of CD31 + area in PDL. ( C ) Representative CD31/vascular endothelial growth factor (VEGF) double immunofluorescence staining images. CD31/VEGF staining without DAPI ( left ) and merged images with DAPI ( right ) are shown. ( D ) Quantitative analysis of VEGF + area in PDL. Scale bars: 100 µm (low magnification), 10 µm (high magnification). Data are mean ± SD (n = 4). ns: not significant, * p < 0.05; ** p < 0.01; *** p < 0.001; Welch’s ANOVA with Games–Howell post hoc test. PDL: periodontal ligament, AB: alveolar bone.

Article Snippet: CD31 , Endothelial cell marker , Bioss Antibodies (Shanghai, China) , bs-0195R , BF647 , 1:100.

Techniques: Immunofluorescence, Staining, Double Immunofluorescence Staining

Journal: Biology

Article Title: Force-Dependent Presence of Senescent Cells Expressing Vascular Endothelial Growth Factor During Orthodontic Tooth Movement

doi: 10.3390/biology15020187

Figure Lengend Snippet: Vascular endothelial growth factor (VEGF) expression of senescent cells during OTM. ( A ) Representative p16 and VEGF double immunofluorescence staining images. p16/VEGF staining without DAPI ( left ) and merged images with DAPI ( right ) are shown. ( B ) Quantitative analysis of p16 + VEGF + area in PDL. ( C ) Schematic illustration of the quantitative analysis for p16 + and VEGF + . ( D ) Quantitative analysis of p16 + VEGF + /VEGF + area in PDL. ( E ) Quantitative analysis of p16 + VEGF + /p16 + area in PDL. Scale bars: 100 µm (low magnification), 10 µm (high magnification). Data are mean ± SD (n = 4). ns; not significant, ** p < 0.01; *** p < 0.001; **** p < 0.0001; one-way ANOVA with Tukey’s test for ( B , D ), and Welch’s ANOVA with Games–Howell post hoc test for ( E ). VEGF: vascular endothelial growth factor, PDL: periodontal ligament, AB: alveolar bone.

Article Snippet: CD31 , Endothelial cell marker , Bioss Antibodies (Shanghai, China) , bs-0195R , BF647 , 1:100.

Techniques: Expressing, Double Immunofluorescence Staining, Staining

Journal: Biology

Article Title: Force-Dependent Presence of Senescent Cells Expressing Vascular Endothelial Growth Factor During Orthodontic Tooth Movement

doi: 10.3390/biology15020187

Figure Lengend Snippet: Cellular senescence of vascular endothelial cells (ECs) induced by OTM. ( A ) Representative p21 and CD31 immunofluorescence staining images. p21/CD31 staining without DAPI ( left ) and merged images with DAPI ( right ) are shown. ( B ) Quantitative analysis of p21 + CD31 + area in PDL. ( C ) Representative p16 and CD31 immunofluorescence staining images. p16/CD31 staining without DAPI ( left ) and merged images with DAPI ( right ) are shown. ( D ) Quantitative analysis of p16 + CD31 + area in PDL. Scale bars: 100 µm (low magnification), 10 µm (high magnification). Data are mean ± SD (n = 4). ns: not significant, * p < 0.05; ** p < 0.01; Kruskal–Wallis with Dunn’s post hoc test. PDL: periodontal ligament, AB: alveolar bone.

Article Snippet: CD31 , Endothelial cell marker , Bioss Antibodies (Shanghai, China) , bs-0195R , BF647 , 1:100.

Techniques: Immunofluorescence, Staining

Journal: Biology

Article Title: Force-Dependent Presence of Senescent Cells Expressing Vascular Endothelial Growth Factor During Orthodontic Tooth Movement

doi: 10.3390/biology15020187

Figure Lengend Snippet: Overlapping analysis of senescent and endothelial markers during OTM. ( A ) Schematic illustration of the overlapping analysis for p21 and CD31 in PDL. ( B ) Quantitative analysis of p21 + CD31 + area/p21 + area. ( C ) Quantitative analysis of p21 + CD31 + area/CD31 + area. ( D ) Schematic illustration of the overlapping analysis for p16 and CD31 in PDL. ( E ) Quantitative analysis of p16 + CD31 + area/p16 + area. ( F ) Quantitative analysis of p16 + CD31 + area/CD31 + area. Data are mean ± SD (n = 4). ns: not significant, * p < 0.05; ** p < 0.01; *** p < 0.001; Welch’s ANOVA with Games–Howell post hoc test for ( B ), one-way ANOVA with Tukey’s test for ( C , F ), and Kruskal–Wallis with Dunn’s post hoc test for ( E ).

Article Snippet: CD31 , Endothelial cell marker , Bioss Antibodies (Shanghai, China) , bs-0195R , BF647 , 1:100.

Techniques:

Journal: Biology

Article Title: Force-Dependent Presence of Senescent Cells Expressing Vascular Endothelial Growth Factor During Orthodontic Tooth Movement

doi: 10.3390/biology15020187

Figure Lengend Snippet: VEGF-expressing senescent ECs. ( A ) Representative p16, CD31, and VEGF triple immunofluorescence staining images. p16/CD31/VEGF staining without DAPI ( left ) and merged images with DAPI ( right ) are shown. ( B ) Quantitative analysis of p16 + CD31 + VEGF + area in PDL. ( C ) Schematic illustration of the quantitative analysis for p16, CD31, and VEGF in PDL. ( D ) Quantitative analysis of p16 + CD31 + VEGF + area/p16 + CD31 + area. ( E ) Quantitative analysis of p16 + CD31 + VEGF + area/p16 + VEGF + area. Scale bars: 100 µm (low magnification), 10 µm (high magnification). Data are mean ± SD (n = 4). ns: not significant, ** p < 0.01; **** p < 0.0001; Kruskal–Wallis with Dunn’s post hoc test for ( B ), and one-way ANOVA with Tukey’s test for ( D , E ). VEGF: vascular endothelial growth factor, PDL: periodontal ligament, AB: alveolar bone.

Article Snippet: CD31 , Endothelial cell marker , Bioss Antibodies (Shanghai, China) , bs-0195R , BF647 , 1:100.

Techniques: Expressing, Immunofluorescence, Staining

Journal: Angiogenesis

Article Title: High-throughput differentiation of human blood vessel organoids reveals overlapping and distinct functions of the cerebral cavernous malformation proteins

doi: 10.1007/s10456-025-09985-5

Figure Lengend Snippet: High-throughput (HT)-compatible and nearly xeno-free synthesis of vascular networks and blood vessel organoids from fluorescently tagged human induced pluripotent stem cells (hiPSCs). A Schematic illustration of the new differentiation protocol and representative images for the main differentiation steps (scale bars: d-2, d0, d3, d5 = 100 µm; d7, d12 = 250 µm; d14, d17 = 500 µm). Created in BioRender. Skowronek, D. (2025) https://BioRender.com/e70e302 . B Shown are the steps of embedding the vascular aggregates in an Akura 96-well plate and transferring the vascular networks from the Akura 96-well plate to a PrimeSurface 96 Slit-well plate. C The use of PrimeSurface 96 Slit-well plates reduces the time required for medium exchange (left image). Akura 96-well plates allow aggregates to be embedded in small cavities, minimizing the matrix surrounding the vascular networks (black arrows) and allowing direct transfer of vascular networks (white arrows) to new plates without time-consuming manual extraction of the networks from the gel (middle and right images). D The new protocol is simple to handle and achieves high synthesis efficiency after minimal training. Shown are the efficiencies of three training runs. E The sprouting efficiency is maintained when fetal bovine serum (FBS) is replaced with human platelet lysate (hPL) or chemically defined Panexin CD (PCD). The total numbers of sufficiently sprouted networks and vascular aggregates with insufficient sprouting are written inside the bars. F,G HiPSC-derived vascular networks (F) and blood vessel organoids (G) differentiated with the HT-compatible and nearly xeno-free protocol consist of a complex network of endothelial cells (CD31) and associated pericytes (PDGFR-β) [representative images; scale bars: 50 µm ( F ); 200 µm ( G )]. White arrowheads indicate angiogenic sprouts. H Perfusion of vascular networks with TMR-amino-dextran in OrganoPlate graft plates shows anastomoses between the GFP-labeled vascular networks and the HUVEC-derived vascular bed (top, white arrowheads) as well as correct formation and permeability of the vascular networks (bottom, scale bar: 50 µm)

Article Snippet: To confirm endothelial differentiation, cells were fixed with 4% PFA at passage 1 and stained for the endothelial markers CD31 (Cell Signaling, 3528S, 1:800), VE-cadherin (2500S, 1:400, Cell Signaling) and VWF (Thermo Fisher Scientific, MA5-14029, 1:66).

Techniques: High Throughput Screening Assay, Transferring, Extraction, Derivative Assay, Labeling, Permeability

Journal: Angiogenesis

Article Title: High-throughput differentiation of human blood vessel organoids reveals overlapping and distinct functions of the cerebral cavernous malformation proteins

doi: 10.1007/s10456-025-09985-5

Figure Lengend Snippet: Perfusion of blood vessel organoids (BVO) on chorioallantoic membranes (CAM). A Schematic illustration of the perfusion approach. Created in BioRender. Skowronek, D. (2025) https://BioRender.com/n06n764 . B Shown are blood vessel organoids cultivated on the CAM associated with chicken blood vessels. The pictures were taken on day 1 and day 6 (scale bars = 2 mm). C Sectioning and H&E staining demonstrated nucleated chicken erythrocytes within the vascular structures of the blood vessel organoid (black arrow head) (upper scale bar = 500 µm; bottom scale bar = 25 µm). D The expression of CD31 (upper image) and PDGFR-β (lower image) were verified by immunohistochemistry staining (brown) (scale bar = 100 µm)

Article Snippet: To confirm endothelial differentiation, cells were fixed with 4% PFA at passage 1 and stained for the endothelial markers CD31 (Cell Signaling, 3528S, 1:800), VE-cadherin (2500S, 1:400, Cell Signaling) and VWF (Thermo Fisher Scientific, MA5-14029, 1:66).

Techniques: Staining, Expressing, Immunohistochemistry

Journal: Angiogenesis

Article Title: High-throughput differentiation of human blood vessel organoids reveals overlapping and distinct functions of the cerebral cavernous malformation proteins

doi: 10.1007/s10456-025-09985-5

Figure Lengend Snippet: Structural defects in KO vascular networks. A Immunofluorescence staining for CD31 (endothelial marker, green) and PDGFR-β (pericyte marker, red) indicated a reduced correlation between ECs and pericytes in CCM1, CCM2, and CCM3 KO vascular networks (scale bar: 100 µm). Correlation was evaluated by using the Pearson’s correlation coefficient (r) calculated with the JACoP ImageJ plugin. B , C Immunofluorescence staining for ZO-1 (B) and VE-cadherin ( C ) revealed irregular tight and adherens junctions in CCM1 , CCM2, and CCM3 KO vascular networks (Alexa 647; scale bars: 25 µm). Statistical analyses demonstrated a significant reduction of Alexa 647 (ZO-1) fluorescence intensity in CCM2 and CCM3 KO networks. Data are presented as individual data points and means. Multiple two-sample t-tests with Welch's correction and Holm-Šídák adjustment for multiple testing were used for statistical analyses (* = Padj < 0.05)

Article Snippet: To confirm endothelial differentiation, cells were fixed with 4% PFA at passage 1 and stained for the endothelial markers CD31 (Cell Signaling, 3528S, 1:800), VE-cadherin (2500S, 1:400, Cell Signaling) and VWF (Thermo Fisher Scientific, MA5-14029, 1:66).

Techniques: Immunofluorescence, Staining, Marker, Fluorescence