Journal: Advanced Science
Article Title: DEL‐1 is an Endogenous Senolytic Protein that Inhibits Senescence‐Associated Bone Loss
doi: 10.1002/advs.202509263
Figure Lengend Snippet: DEL‐1 deficiency exacerbates chemotherapy‐induced bone loss. Doxorubicin was administered to 9‐week‐old WT and Del1 KO mice at a dose of 5 mg kg −1 per week via intraperitoneal injection for 4 weeks. After 4 weeks, femurs from both WT and Del1 KO mice were extracted and analyzed using microcomputed tomography to assess bone loss. A) Schematic representation of the experimental timeline for doxorubicin administration. B) Representative µCT images of bone microarchitecture at the sagittal plane (left) and the mid‐diaphysis of the femur (middle). Quantification of µCT‐derived cortical thickness (Ct.Th(mm), right). C) Representative µCT images of bone microarchitecture at the distal femoral metaphysis (left). Quantification of µCT–derived trabecular bone parameters, including bone volume fraction (Tb.BV/TV(%)), trabecular number (Tb.N(mm −1 )), trabecular thickness (Tb.Th(mm)), and trabecular space (Tb.Sp(mm)) (right). D,E) Fasting serum levels of bone formation marker osteocalcin (D), bone resorption markers collagen type I cross‐linked C‐telopeptide (CTX1) and tartrate‐resistant acid phosphatase 5b (TRAcP 5b) (E). F) Histology, immunostaining, and TRAP staining of femurs. Representative images of H&E staining (scale bar, 500 µm), TRAP‐positive osteoclasts (purple; scale bar, 100 µm), and osteocalcin (OCN)‐positive osteoblasts (green; scale bar, 100 µm) in a boxed area on the H&E staining panels. The nuclei were counterstained using DAPI (blue). G) Expression of senescence‐related genes, p21, p53, Il6, Cxcl2, Tgfβ1, and p16 , in the bone marrow cells assessed by qPCR. Gapdh was used for normalization of mRNA expression and the gene expression of WT mice was assigned an average value of 1. H) Levels of IL‐6, TGFβ1, and CXCL2 in bone marrow fluid, measured by ELISA. I) Representative fluorescence images of p21⁺ cells (purple) in the bone marrow compartment of femoral bone sections. The nuclei were counterstained with DAPI (blue). Scale bar, 100 µm. Data are means ± SD (B,C: n =4‐5 mice/group; D,E: n =5 mice/group; G: n =3–4 mice/group; H: n =6–7 mice/group). * p < 0.05; ** p < 0.01; *** p <0.001; NS, not significant. Unpaired Student's t ‐test, except for “CTX1” in panel E (Mann‐Whitney U test).
Article Snippet: Serum levels of CTX1 (catalogue no. AC‐06F1, Immunodiagnostic Systems), TRAcP 5b (catalogue no. SB‐TR103, Immunodiagnostic Systems), and osteocalcin (catalogue no. EEL003, Thermo Fisher Scientific), were measured using ELISA kits, following the manufacturer's protocol.
Techniques: Injection, Tomography, Derivative Assay, Marker, Immunostaining, Staining, Expressing, Gene Expression, Enzyme-linked Immunosorbent Assay, Fluorescence, MANN-WHITNEY