Journal: Glycobiology

Article Title: Comprehensive analysis of the skeletal phenotype in Chst14 −/− mice: implications for dermatan sulfate in bone structure and strength

doi: 10.1093/glycob/cwag037

Figure Lengend Snippet: Alterations in bone metabolic factors in Chst14 −/− mice. A) Relative mRNA levels of bone gamma-carboxyglutamate protein ( Bglap ) and acid phosphatase 5 ( Acp5 ) in femurs of WT (+/+) and Chst14 −/− (−/−) mice at 12 and 52 weeks old (mean ± SD; n = 6 for 12-week-old WT and Chst14 −/− mice and for 52-week-old WT mice; n = 5 for 52-week-old Chst14 −/− mice). B) Relative mRNA levels of receptor activator of NF-κB ( Rank ), receptor activator of NF-κB ligand ( Rankl ), and osteoprotegerin ( Opg ) in femurs of WT (+/+) and Chst14 −/− (−/−) mice at 12 and 52 weeks old (mean ± SD; n = 6 for 12-week-old WT and Chst14 −/− mice and for 52-week-old WT mice; n = 5 for 52-week-old Chst14 −/− mice). Two-way ANOVA followed by Tukey’s post hoc test. C) Relative mRNA levels of cathepsin K ( Ctsk ) in femurs of WT (+/+) and Chst14 −/− (−/−) mice at 12 and 52 weeks old (mean ± SD; n = 6 for 12-week-old WT and Chst14 −/− mice and for 52-week-old WT mice; n = 5 for 52-week-old Chst14 −/− mice). D) Serum concentrations of tartrate-resistant acid phosphatase 5b (TRACP-5b), a marker of bone resorption (mean ± SD, each group: n = 6). In Chst14 −/− mice, gene expression analysis of cortical bone showed age-dependent changes in bone formation and resorption markers, as well as increased expression of an osteoclast differentiation factor. By contrast, the serum osteoclast marker did not change. Statistical analyses for panels A–D were performed using two-way ANOVA followed by Tukey’s post hoc test. Statistical significance is shown only for biologically relevant comparisons (+/+ vs −/− at the same age and 12 vs 52 weeks within genotype). Complete results are provided in – . Statistical significance in the graphs (A–D) is indicated as follows: * P < 0.05, ** P < 0.01; no asterisk indicates no statistically significant difference.

Article Snippet: Serum Ca, P i , estradiol, and TRACP-5b were measured by Oriental Yeast Co., Ltd. (Tokyo, Japan).

Techniques: Marker, Gene Expression, Expressing

Journal: Journal of Bone and Mineral Research

Article Title: New lens on congenital mild bone fragility: a novel Col1a1 knockout mouse model for osteogenesis imperfecta type 1

doi: 10.1093/jbmr/zjaf138

Figure Lengend Snippet: Altered collagen gene expression and serum biomarkers in hiOI mice. (A) Serum P1NP levels were significantly reduced in hiOI mice at 8 and 24 wk of age. (B) Serum TRAcP 5b levels remained normal in hiOI mice. (C-D) Real-time quantitative PCR analysis of Col1a1 and Col1a2 mRNA expression and their ratio in 20 hiOI and 20 WT mice at 8 wk (C) and 24 wk of age (D). (E) Serum P1NP levels were positively correlated with the Col1a1/Col1a2 mRNA expression ratio in bone at 8 wk ( p = .0088, r = 0.4137) and 24 wk ( p < .0001, r = 0.6246). (F) The Col1a1/Col1a2 ratio was correlated with trabecular and cortical bone volume in 8-wk-old mice but not in 24-wk-old mice. p -values are indicated as follows: ≤.05 ( * ), ≤.01 ( ** ), ≤.001 ( *** ), ≤.0001 ( **** ). Data shown as mean ± SD. All comparisons between WT and hiOI data were performed using an unpaired t -test. Gender subgroups were analyzed using a two-way ANOVA with a main effect and multiple comparisons. Correlations between P1NP levels, collagen mRNA expression ratios, and bone volumes were assessed using Pearson correlation analysis.

Article Snippet: Additionally, the serum levels of tartrate-resistant acid phosphatase (TRAcP) 5b (SB-TR103, Immunodiagnostic systems) were analyzed in 8-wk-old mice to assess bone resorption.

Techniques: Gene Expression, Real-time Polymerase Chain Reaction, Expressing

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