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: The serum levels of procollagen type I N-terminal propeptide (P1NP) (P1NP EIA kit, AC-33F1, Immunodiagnostic systems), a bone formation marker, were assessed in 8-wk-old and 24-wk-old mice by ELISA according to the manufacturer’s protocol.

Techniques: Gene Expression, Real-time Polymerase Chain Reaction, 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: Reduced bone collagen expression in hiOI mice. (A) Western blot analysis of α1(I) and α2(I) collagen protein bands extracted from ulnae bones of WT ( n = 3) and hiOI ( n = 3) mice. (B) Quantification showing the percentage reduction of α1(I), α2(I), total collagen type I, and the Col1a1 / Col1a2 protein ratio in hiOI relative to WT ulnae. (C) Genotype, Col1a1 / Col1a2 mRNA ratio, serum P1NP levels, and total collagen type I (normalized to tissue weight) in ulnae. (D) A positive correlation between serum P1NP levels and total collagen type I measured by western blot is shown. Statistical significance indicated by * for p ≤ .05.

Article Snippet: The serum levels of procollagen type I N-terminal propeptide (P1NP) (P1NP EIA kit, AC-33F1, Immunodiagnostic systems), a bone formation marker, were assessed in 8-wk-old and 24-wk-old mice by ELISA according to the manufacturer’s protocol.

Techniques: Expressing, Western Blot

Journal: BioMed Research International

Article Title: The Effects of Neonatal Zingerone Administration and Adolescent Alcohol Exposure on Bone Health Markers and Morphometry in Male Sprague–Dawley Rats

doi: 10.1155/bmri/7060593

Figure Lengend Snippet: Effects of neonatally administered zingerone (40 mg/kg/day) and adolescent alcohol (1 g/kg) on plasma concentrations of (A) BALP, (B) OC, and (C) P1NP as bone health markers. Data were presented as mean ± standard deviation. BALP, OC, and P1NP levels were elevated in the alcohol‐treated groups with or without zingerone. Statistically significant if ∗ p ≤ 0.05, ∗∗ p ≤ 0.01, ∗∗∗ p ≤ 0.001, ∗∗∗∗ p ≤ 0.0001 between treatment groups. Group 1 ( C ) received no treatment in the neonatal phase and during adolescence; Group 2 ( Z + W ) was treated with zingerone (40 mg/kg) in the neonatal phase and water during adolescence; Group 3 ( A + A ) treated with alcohol during both the neonatal and adolescent phase (1 g/kg ethanol); Group 4 ( Z + A ) treated with zingerone in the neonatal phase and alcohol in adolescence; Group 5 (ZA + A ) treated with a combination of zingerone–alcohol in the neonatal phase, followed by alcohol in adolescence. A —alcohol; C —control; W —water; Z —zingerone; ZA—zingerone‐alcohol; BALP—bone‐specific alkaline phosphatase, OC—osteocalcin, P1NP—procollagen Type 1 N‐terminal propeptide.

Article Snippet: The plasma activity of BALP (Catalog No. ER0761, FineTest, Wuhan, China), OC (Catalog No. ER1205, FineTest, Wuhan, China), and P1NP (Catalog No. E‐EL‐R1414, Elabscience, Houston, Texas, United States) were determined in rat plasma as described by the manufacturer′s instructions.

Techniques: Clinical Proteomics, Standard Deviation, Control

Journal: Bone Research

Article Title: Spatially resolved osteoblast-traced transcriptomics uncovers TGF-β as a combination target with sclerostin in osteoporosis

doi: 10.1038/s41413-026-00521-9

Figure Lengend Snippet: Lineage tracing reveals that TGF-β regulates osteoblast-BLC conversion and reactivation. a Experimental timeline of tamoxifen pulse and TGF-β/TGFβ-Ab administration in Dmp1 -CreERt2:mTmG mice. b Representative confocal images of femoral periosteum showing GFP (green), tdTomato (red) signals, and DAPI (blue). Scale bar = 20 μm. c Violin plot of GFP+ cell thickness. Each point represents individual cells. The Wilcoxon rank-sum test was used for statistics. d Bar plot of GFP+ cells per unit length. Cell numbers were determined from three comparable sections per mouse, with eight fields (400× magnification) analyzed per section. The Wilcoxon rank-sum test was used for statistics. e Box plot of serum P1NP levels. A t -test was used for statistics. Group sizes: 8 weeks ( n = 6), 9 weeks ( n = 5), TGF-β ( n = 4), Inactive (12 weeks control + 13 weeks control; n = 6), Scl-Ab+TGF-β ( n = 3), Scl-Ab ( n = 3), TGFβ-Ab ( n = 4), and Scl-Ab+TGFβ-Ab ( n = 5). * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.000 1; ns not significant

Article Snippet: Serum levels of P1NP, CTx1, TRAcP, OPG, RANKL, and TGF-β1 were measured via an enzyme-linked immunosorbent assay (ELISA) performed using ELISA kit for mouse P1NP (AC-33F1, Immunodiagnostic Systems, UK), mouse CTx1 (AC-06F1, Immunodiagnostic Systems, UK), mouse TRAcP 5b (SB-TR103, Immunodiagnostic Systems, UK), mouse OPG (MOP00, R&D systems, USA), mouse RANKL (MTR00, R&D systems, USA), and mouse TGF-β1 (DY1679-05 and DY007B, R&D systems, USA) according to the manufacturer’s instruction.

Techniques: Control

Journal: Bone Research

Article Title: Spatially resolved osteoblast-traced transcriptomics uncovers TGF-β as a combination target with sclerostin in osteoporosis

doi: 10.1038/s41413-026-00521-9

Figure Lengend Snippet: Dual inhibition of TGF-β and sclerostin increases bone mass in a hindlimb unloading model. a Experimental timeline of hindlimb unloading using C57B1/6 J mice and treatment with TGFβ-Ab, Scl-Ab, or both. Mice euthanized at week 12. b Representative μCT images of femoral diaphysis. c Representative confocal images of trabecular bone showing calcein (green), alizarin (red) signals, and DAPI (blue). Scale bar = 100 μm. d Representative images of TRAP-stained femur sections; violet = TRAP-positive, green = counterstain. Scale bar = 200 μm. e – h Bar plot of trabecular bone volume/total volume (Tra. BV/TV, e ), trabecular thickness (Tb. Th, f ), trabecular separation (Tb. Sp, g ), and trabecular number (Tb. N, h ) from μCT. Data = mean ± standard deviation. i , j Bar plot of mineral apposition rate (Tra. MAR, i ) and trabecular bone formation rate per bone surface (Tra. BFR/BS, j ) from labeling images. Data = mean ± standard deviation k Box plot of TRAP+ osteoclasts per bone surface (OC.N/BS), measured through ImageJ from TRAP-stained slides. l – n Box plot of serum P1NP ( l ) and TRAP ( m ) levels, and RANKL/OPG ratio ( n ). Group sizes: Control ( n = 5), Un_con ( n = 5), Scl-Ab ( n = 6), TGFβ-Ab ( n = 6), and Scl-Ab+TGFβ-Ab ( n = 7). Each point represents an individual mouse. ANOVA was used for statistics; * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.000 1; ns not significant

Article Snippet: Serum levels of P1NP, CTx1, TRAcP, OPG, RANKL, and TGF-β1 were measured via an enzyme-linked immunosorbent assay (ELISA) performed using ELISA kit for mouse P1NP (AC-33F1, Immunodiagnostic Systems, UK), mouse CTx1 (AC-06F1, Immunodiagnostic Systems, UK), mouse TRAcP 5b (SB-TR103, Immunodiagnostic Systems, UK), mouse OPG (MOP00, R&D systems, USA), mouse RANKL (MTR00, R&D systems, USA), and mouse TGF-β1 (DY1679-05 and DY007B, R&D systems, USA) according to the manufacturer’s instruction.

Techniques: Inhibition, Staining, Standard Deviation, Labeling, Control

Journal: Bone Research

Article Title: Expansion of bone marrow adipocytes in obese mice leads to PD-L1-driven bone marrow immunosuppression and osteoclastogenesis

doi: 10.1038/s41413-026-00509-5

Figure Lengend Snippet: Obesity parameters and metabolic phenotype in male B6 mice. a Body weight (g) measurements over 12 weeks for LFD and HFD-fed male B6 mice ( n = 32-41). b Change in weight (%) (dotted line at 28% represents parameter for LFD and dotted line at 58% represents parameter for HFD). c Fold change in fat mass (g) (dotted line at 1.0 represents parameter for LFD and dotted line at 3.3 represents parameter for obese-HFD). d Pearson correlation analysis showing that the change in weight or fat mass fold change both negatively correlate to trabecular bone loss. e Correlation matrix that shows the change in weight or fat mass fold change both negatively correlate to trabecular bone loss (vertical line and horizontal line represent obesity cutoff). f Glucose tolerance test (GTT) for LFD ( n = 26), obese HFD-fed (OB-HFD; n = 34) and non-obese HFD-fed (NO-HFD; n = 7). g Insulin tolerance test (ITT) for LFD, obese HFD-fed (OB-HFD), and non-obese HFD-fed (NO-HFD). h Serum adiponectin levels ( n = 7). i Serum leptin levels. j Serum procollagen type I N-propeptide (P1NP) levels. k Serum tartrate-resistant acid phosphatase 5b (TRAcP 5b) levels. Analyses for a , f , and g were performed as 2-way ANOVA with Šídák’s multiple comparisons test. Significance for the post-hoc analysis for f , g was defined as: * P < 0.05 LFD vs. OB-HFD, # P < 0.05 LFD vs. NO-HFD-fed, and $ P < 0.05 OB-HFD vs. NO - HFD-fed. Analyses for h , i and k were performed as a Kruskal-Wallis test with Dunn’s multiple comparison. Analysis for j was performed as a One-way ANOVA with Tukey’s multiple comparisons test

Article Snippet: Serum protein analysis of adiponectin (Mouse Adiponectin/Acrp30 Quantikine ELISA, R&D Systems MRP300), leptin (Mouse/Rat Leptin Quantikine ELISA, R&D Systems MOB00B), TRAP (Mousetrap TRAcP 5b ELISA, Immunodiagnostic Systems SB-TR103), CTX-1 (Mouse CTX ELISA Kit, Immunodiagnostic Systems AC-06F1), and P1NP (Rat/Mouse P1NP EIA, Immunodiagnostic Systems AC-33F1) were measured by ELISA according to manufacturer’s instructions.

Techniques: Comparison

Journal: Bone Research

Article Title: Expansion of bone marrow adipocytes in obese mice leads to PD-L1-driven bone marrow immunosuppression and osteoclastogenesis

doi: 10.1038/s41413-026-00509-5

Figure Lengend Snippet: BMAT depletion in obese mice improved BM immunosuppression through a decrease in PD-L1 + myeloid cells. a Number of BM adipocytes within the proximal tibia (BM.Ads/Ma.AR, number/mm 2 ; n = 10-13). b Average size of the BMAT within the proximal tibia (μm 2 ). c Representative H&E images of the BMAT within the proximal tibia (scale bar = 250 μm). d Immune and inflammatory gene expression of BM adipocytes from LFD mice relative to TBP ( n = 8; represented as the mean with individual data points). e Immune and inflammatory gene expression of BM adipocytes from OB-HFD mice relative to TBP ( n = 8; represented as the mean with individual data points). f F4/80 IHC of gWAT (red arrows indicate minimal macrophage infiltration in LFD-fed mice). g Number of PD-L1 + myeloid cells. h Number of PD-1 + osteoclast precursor. i Serum procollagen type I N-propeptide (P1NP) levels ( n = 8). j Serum C-terminal telopeptide of type 1 collagen (CTX-1), a bone resorption marker. k Serum TRAcP 5b levels. Analyses for a , b and g – k were performed as 2-way ANOVA with uncorrected Fisher’s LSD. Analyses for d , e were performed as multiple unpaired t -tests and multiple Mann-Whitney tests, respectively

Article Snippet: Serum protein analysis of adiponectin (Mouse Adiponectin/Acrp30 Quantikine ELISA, R&D Systems MRP300), leptin (Mouse/Rat Leptin Quantikine ELISA, R&D Systems MOB00B), TRAP (Mousetrap TRAcP 5b ELISA, Immunodiagnostic Systems SB-TR103), CTX-1 (Mouse CTX ELISA Kit, Immunodiagnostic Systems AC-06F1), and P1NP (Rat/Mouse P1NP EIA, Immunodiagnostic Systems AC-33F1) were measured by ELISA according to manufacturer’s instructions.

Techniques: Gene Expression, Marker, MANN-WHITNEY

Journal: Journal of Pharmaceutical Analysis

Article Title: MST4 as a key driver of osteoclast activation in osteoporosis

doi: 10.1016/j.jpha.2025.101401

Figure Lengend Snippet: Upregulation of mammalian Sterile 20-like kinase 4 (MST4) expression in peripheral blood mononuclear cells of clinical osteoporosis patients (OP). (A) Quantitative real-time polymerase chain reaction (RT-qPCR) detection of MST4 mRNA levels in peripheral blood mononuclear cells of OP and healthy controls (Control) ( n = 6). (B, C) Western blot analysis of MST4 protein expression levels (B) and corresponding statistics (C) in OP and Control ( n = 6). (D) Bone mineral density (BMD) levels in OP and Control. (E, F) Expression levels of bone formation markers osteocalcin (OC) (E) and procollagen type I N -terminal propeptide (P1NP) (F) in OP and Control. (G, H) Expression levels of bone formation markers β-CrossLaps (β-CTX) and tartrate-resistant acid phosphatase 5b (TRAcP 5b) in OP and Control. (I) Receiver operating characteristic (ROC) curve analysis of MST4 expression for predicting fragility fractures in OP ( n = 60). ∗ P < 0.05 compared between groups, n = 60 per group（OP 60，Control 60); ns: not significant.

Article Snippet: Bone metabolism markers in peripheral blood, such as the bone formation marker procollagen type I N -terminal propeptide (P1NP) (D711084-0048; Sangon Biotech Co., Ltd., Shanghai, China), osteocalcin (OC) (D711187-0048; Sangon Biotech Co., Ltd., Shanghai, China), and bone resorption markers like β-CrossLaps (β-CTX) (D711348; Sangon Biotech Co., Ltd., Shanghai, China), and tartrate-resistant acid phosphatase 5b (TRAcP 5b) (P0332; Beyotime Biotechnology, Shanghai, China), were measured using enzyme-linked immunosorbent assay (ELISA).

Techniques: Sterility, Expressing, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Control, Western Blot