trap staining  (Servicebio Inc)

Journal: Dose-Response

Article Title: Skeletal Stem Cells Rescue Radiation-Induced Osteogenic Precursor Cell Dysfunction via the Wnt/β-Catenin Signaling Pathway

doi: 10.1177/15593258261440983

Figure Lengend Snippet: SSCs alleviate the radiation-induced bone injury in mice. (A–G) Micro-CT analysis of bone microstructure. (A) Representative micro-CT images of femurs. Quantitative analysis of (B) bone mineral density (BMD), (C) bone volume fraction (BV/TV), (D) trabecular thickness (Tb.Th), (E) trabecular number (Tb.N), (F) connectivity density (Conn.D), and (G) trabecular separation (Tb.Sp) at 2- and 4-weeks post irradiation. (H–K) Histological analysis (Scale bar: 100 μm). (H) H&E staining showing steatosis (arrows) and (I) quantitative analysis of steatotic lesions per field. (J) TRAP staining showing osteoclasts (arrows) and (K) quantitative analysis of osteoclast number per field. (L–O) Immunohistochemical staining of osteogenic markers (Scale bar: 100 μm). (L) Osterix staining and (M) quantitative analysis of Osterix-positive area. (N) β-catenin staining and (O) quantitative analysis of β-catenin-positive area. All experiments were conducted in three groups: Control, irradiation (IR), and IR plus SSC (IR+SSC) at 2- and 4-weeks post-irradiation. All data are presented as mean ± SD, with statistical significance determined by unpaired two-tailed Student’s t-test (* p < 0.05; ** p < 0.01; *** p < 0.001)

Article Snippet: Paraffin sections of femurs were dewaxed to water, and TRAP staining was performed using a TRAP staining kit (Servicebio, Wuhan, China, G1050-50T) according to the manufacturer’s instructions.

Techniques: Micro-CT, Irradiation, Staining, Immunohistochemical staining, Control, Two Tailed Test

Journal: Bone & Joint Research

Article Title: OX40L in endothelial cells promotes temporomandibular joint subchondral bone angiogenesis and osteoclastogenesis in mice

doi: 10.1302/2046-3758.154.BJR-2025-0249.R1

Figure Lengend Snippet: Changes in subchondral vascular proliferation and bone remodelling during different progression cycles of temporomandibular joint osteoarthritis (TMJOA). TMJOA was induced in C57BL/6J mice using a unilateral anterior crossbite (UAC) model, and subsequently assessed at two, four, six, and eight weeks after initiation. a) Haematoxylin & eosin (HE) staining in mouse condylar cartilage. The double arrow shows the thickness of the cartilage layer. Scale bar: 100 µm. b) Safranin O/fast green staining in mouse condylar cartilage. Scale bar: 100 µm. c) Tartrate-resistant acid phosphatase (TRAP) staining in mouse condylar subchondral bone. Scale bar: 100 µm. d) Micro-CT image in mouse condyle. e) CD31 immunofluorescence staining in mouse condylar subchondral bone. Scale bar: 100 µm. DAPI, 4′,6-diamidino-2-phenylindole.

Article Snippet: Additionally, sagittal sections were processed for tartrate-resistant acid phosphatase (TRAP) staining (Servicebio, China) to quantify osteoclast numbers in the subchondral bone.

Techniques: Staining, Micro-CT, Immunofluorescence

Journal: Bone & Joint Research

Article Title: OX40L in endothelial cells promotes temporomandibular joint subchondral bone angiogenesis and osteoclastogenesis in mice

doi: 10.1302/2046-3758.154.BJR-2025-0249.R1

Figure Lengend Snippet: Abnormal activation of OX40L in endothelial cells can promote angiogenesis and osteoclast induction. Human bone microvascular endothelial cells were pretreated with KY1005 for six hours, followed by treatment with sOX40 for 48 hours. a) Detection of vascular endothelial growth factor (VEGF) gene expression using quantitative polymerase chain reaction (qPCR). b) Detection of VEGF protein expression using western blotting. c) Representative images of endothelial cell scratch experiments. Scale bar: 100 µm. d) Representative images of endothelial cell tubule formation experiments. Scale bar: 100 µm. e) Representative image of tartrate-resistant acid phosphatase (TRAP) staining after endothelial cells were co-cultured with RAW264.7 cells. Scale bar: 50 µm. f) Bone resorption pits were shown by scanning electron microscopy. Scale bar: 500 nm.

Article Snippet: Additionally, sagittal sections were processed for tartrate-resistant acid phosphatase (TRAP) staining (Servicebio, China) to quantify osteoclast numbers in the subchondral bone.

Techniques: Activation Assay, Gene Expression, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Staining, Cell Culture, Electron Microscopy

Journal: Bone & Joint Research

Article Title: OX40L in endothelial cells promotes temporomandibular joint subchondral bone angiogenesis and osteoclastogenesis in mice

doi: 10.1302/2046-3758.154.BJR-2025-0249.R1

Figure Lengend Snippet: Blocking the OX40–OX40L pathway alleviates abnormal subchondral angiogenesis and osteoclastic activity in early temporomandibular joint osteoarthritis (TMJOA). After four weeks of unilateral anterior cross-bite (UAC) modeling, C57BL/6J mice received intraperitoneal injections of OX40L monoclonal antibody. a) CD4 immunofluorescence staining in mouse condylar subchondral bone. Scale bar: 100 µm. b) Statistical analysis of the proportion of CD4 + T cells in in mouse condylar subchondral bone. c) CD31 immunofluorescence staining in mouse condylar subchondral bone. Scale bar: 100 µm. d) Statistical analysis of blood vessel area in mouse condylar subchondral bone. e) Tartrate-resistant acid phosphatase (TRAP) staining in mouse condylar subchondral bone. Scale bar: 100 µm. f) Statistical analysis of the number of TRAP + osteoclasts in mouse condylar subchondral bone. g) Micro-CT image of mouse condyle. h) to k) Results of morphological analysis of condylar subchondral bone, including statistical analysis of bone volume to tissue volume (BV/TV), trabecular number (Tb.N), trabecular thickness (Tb.Th), and trabecular separation (Tb.Sp). Data are presented as the mean (SD) (n = 3). Statistical significance was determined by independent-samples t -test.*p < 0.05, **p < 0.01 and ***p < 0.001. DAPI, 4′,6-diamidino-2-phenylindole; PBS, phosphate-buffered saline.

Article Snippet: Additionally, sagittal sections were processed for tartrate-resistant acid phosphatase (TRAP) staining (Servicebio, China) to quantify osteoclast numbers in the subchondral bone.

Techniques: Blocking Assay, Activity Assay, Immunofluorescence, Staining, Micro-CT, Saline