Journal: World Journal of Stem Cells

Article Title: EZH2, via an association with KDM2B, modulates osteogenic differentiation of root apical papillary stem cells

doi: 10.4252/wjsc.v17.i4.103482

Figure Lengend Snippet: Knockdown of enhancer of zeste homolog 2 inhibits steo/dentinogenic differentiation potential of human apical papillary stem cells. A: Quantitative polymerase chain reaction showed that the expression of enhancer of zeste homolog 2 (EZH2) was inhibited in human apical papillary stem cells (hSCAPs); B: Western blot analysis confirmed the knockdown of EZH2 in hSCAPs; C: Knockdown of EZH2 decreased alkaline phosphatase activity in hSCAPs; D and E: Alizarin red staining and quantitative calcium analysis demonstrated that knockdown of EZH2 inhibited mineralization in hSCAPs; F-H: Quantitative polymerase chain reaction showed that knockdown of EZH2 downregulated mRNA expression levels of bone sialoprotein (F), dentin sialophosphoprotein (G), and osteocalcin (H) in hSCAPs. GAPDH and ACTB was used as the internal controls. Data are presented as the mean ± SD ( n = 3). Statistical analysis was performed using Student’s t -test. a P ≤ 0.05, b P ≤ 0.01, c P ≤ 0.001. EZH2: Enhancer of zeste homolog 2; BSP: Bone sialoprotein; DSPP: Dentin sialophosphoprotein; OCN: Osteocalcin.

Article Snippet: Immunohistochemical analysis followed established protocols[ ] using primary antibodies against osteocalcin (OCN) (Bioss bs-4917R) and DSPP (Bioss bs10316R).

Techniques: Knockdown, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Activity Assay, Staining

Journal: World Journal of Stem Cells

Article Title: EZH2, via an association with KDM2B, modulates osteogenic differentiation of root apical papillary stem cells

doi: 10.4252/wjsc.v17.i4.103482

Figure Lengend Snippet: Overexpression of enhancer of zeste homolog 2 enhances osteo/dentinogenic differentiation potential of human apical papillary stem cells. A: Quantitative polymerase chain reaction showed that enhancer of zeste homolog 2 (EZH2) was overexpressed in human apical papillary stem cells (hSCAPs); B: Western blot analysis confirmed overexpression of EZH2 in hSCAPs; C: Overexpression of EZH2 increased alkaline phosphatase activity in hSCAPs; D and E: Alizarin red staining and quantitative calcium analysis results demonstrated that overexpression of EZH2 enhanced mineralization in hSCAPs; F-H: Quantitative polymerase chain reaction showed that overexpression of EZH2 upregulated mRNA expression levels of bone sialoprotein (F), dentin sialophosphoprotein (G), and osteocalcin (H) in hSCAPs; I: Hematoxylin-eosin staining and quantitative measurement showed that overexpression of EZH2 promoted bone/dentin-like tissue formation. Scale bar = 100 μm (B: Bone/dentin-like tissues; HA: Hydroxyapatite tricalcium carrier; CT: Connective tissue); J: Immunohistochemical staining and quantitative analysis of dentin sialophosphoprotein and bone sialoprotein. GAPDH and ACTB were used as the internal controls. Data are presented as the mean ± SD ( n = 3). Statistical analysis was performed using Student’s t -test. a P ≤ 0.05, b P ≤ 0.01, c P ≤ 0.001. EZH2: Enhancer of zeste homolog 2; BSP: Bone sialoprotein; DSPP: Dentin sialophosphoprotein; OCN: Osteocalcin.

Article Snippet: Immunohistochemical analysis followed established protocols[ ] using primary antibodies against osteocalcin (OCN) (Bioss bs-4917R) and DSPP (Bioss bs10316R).

Techniques: Over Expression, Real-time Polymerase Chain Reaction, Western Blot, Activity Assay, Staining, Expressing, Immunohistochemical staining

Journal: Bioactive Materials

Article Title: Pretreated exosomes by electrical stimulation accelerate bone regeneration

doi: 10.1016/j.bioactmat.2025.04.019

Figure Lengend Snippet: Histological staining of the distal femur defect site at 6-weeks post-injection. (A) Hematoxylin and eosin (H&E) staining, (B) Masson's trichrome staining, and (C) Immunohistochemical staining for OCN. D) The relative proportion of collagen from the quantitative analysis of Masson staining. E) Quantitative analysis results of IHC Results are shown as mean ± SD. ns: non-significant differences, ∗∗p < 0.01.

Article Snippet: They were then co-incubated with the primary antibody, specifically the OCN primary antibody from Beyotime, China.

Techniques: Staining, Injection, Immunohistochemical staining

Journal: Materials Today Bio

Article Title: Bi-phasic integrated silk fibroin/polycaprolactone scaffolds for osteochondral regeneration inspired by the native joint tissue and interface

doi: 10.1016/j.mtbio.2025.101737

Figure Lengend Snippet: Biocompatibility and osteogenic differentiation potential of MC3T3-E1 seeded on 2D/3D SP (1:3) /GelSP (1:4) scaffolds. (a) The cell distribution inside 3DSP (1:3) /GelSP (1:4) . Dil-labeled ATDC5(red) were seeded on the SP (1:3) layer, and Dio-labeled MC3T3-E1(green) were seeded on the GelSP (1:4) layer of 3DSP (1:3) /GelSP (1:4) for 7days. (b) Cell proliferation of MC3T3-E1 seeded on 2D/3D SP (1:3) /GelSP (1:4) scaffolds at days 1, 3, and 7, as measured by the CCK-8 assay. (c) The viability of MC3T3-E1 seeded on 2D/3D SP (1:3) /GelSP (1:4) scaffolds by live/dead staining and its 3D fluorescent images for 3 days. (d) Cytoskeleton staining of MC3T3-E1 cultured on different scaffolds for 3 days. (e–g) The relative mRNA expression levels of OPN, Runx2, and OCN of MC3T3-E1 seeded on 2D/3D SP (1:3) /GelSP (1:4) scaffolds for 14 days. (h) Immunofluorescence images of osteocalcin (OCN) in MC3T3-E1 seeded on 2D/3D SP (1:3) /GelSP (1:4) scaffolds. (∗p < 0.05. ∗∗p < 0.01. ∗∗∗p < 0.001).

Article Snippet: After 14 days of in vitro culture, cells/scaffold complexes were fixed in 4 % paraformaldehyde, permeabilized with 0.1 % Triton X-100 for 10 min, and blocked with 3 % BSA for 1 h. The cells were then incubated with the primary antibody against OCN (Proteintech, China, 1:500) and collagen II (Proteintech, China, 1:500) overnight respectively, followed by 1 h co-incubation with Cy3-conjugated fluorescence secondary antibody (Abclonal, China, 1:200).

Techniques: Labeling, CCK-8 Assay, Staining, Cell Culture, Expressing, Immunofluorescence

Journal: Materials Today Bio

Article Title: Bi-phasic integrated silk fibroin/polycaprolactone scaffolds for osteochondral regeneration inspired by the native joint tissue and interface

doi: 10.1016/j.mtbio.2025.101737

Figure Lengend Snippet: Immunological assessment of osteochondral defect in vivo at 6 and 12 weeks. Representative images of COL-II (a, b) and OCN (c, d) immunohistochemical staining (The images on the right are magnified views of the left images, with black arrows indicating the boundary between normal cartilage (N) and repaired cartilage (R), V represents vessel, S represents incompletely degraded scaffolds).

Article Snippet: After 14 days of in vitro culture, cells/scaffold complexes were fixed in 4 % paraformaldehyde, permeabilized with 0.1 % Triton X-100 for 10 min, and blocked with 3 % BSA for 1 h. The cells were then incubated with the primary antibody against OCN (Proteintech, China, 1:500) and collagen II (Proteintech, China, 1:500) overnight respectively, followed by 1 h co-incubation with Cy3-conjugated fluorescence secondary antibody (Abclonal, China, 1:200).

Techniques: In Vivo, Immunohistochemical staining, Staining