competitive cgmp elisa kit  (Cell Signaling Technology Inc)

Journal: Bioactive Materials

Article Title: Shape memory bone screws loading L-arginine and Ca 2+ propagate mechanical stimulation, energize bone cells and augment bone regeneration

doi: 10.1016/j.bioactmat.2025.07.007

Figure Lengend Snippet: Schematics showing the design and application of SMP bone screw. (A) Fabrication and shape memory function of SMP bone screw; (B) Schematic illustration of preparation process of SMP-Ca/ArgX (X = 0.5, 1, 2) bone screw; (C) Dopamine-assisted coating of Ca 2+ and Arg's SMP-Ca/Arg screw and subsequent implantation into the bone defect site; (D) SMP-Ca/Arg-P bone screw (an SMP bone screw coated with PDA, Ca 2+ , and 2.0 mg/mL of Arg, and then compressed) can release mechanical force and Arg/Ca 2+ for coupled osteogenesis–angiogenesis in bone repair; (E) Schematic illustration of the forces activate the mechanosensitive transient receptor potential vanilloid 4 (TRPV4) protein and piezo type mechanosensitive ion channel component 2 (PIEZO2). This activation boosts adenosine triphosphate (ATP) generation and enhancing cellular metabolism, facilitating the influx of Ca 2+ and Arg released by the SMP-Ca/Arg bone screws through activated calcium channels (e.g., TRPV4 and PIEZO2) and cell membrane proteins (e.g., cationic amino acid transporter (CAT)). The elevated Ca 2+ level further upregulates calmodulin (CaM) and increase the activity of nitric oxide synthase (NOS) for Arg decomposition to regenerate nitric oxide (NO). This subsequently activates the NO-cGMP pathway, promoting coupled osteogenesis and angiogenesis. The calcium signaling pathway is visually represented by blue arrows, the NO-cGMP pathway is represented by red arrows, and the forces exerted by the SMP-Ca/Arg-P bone screw is indicated by orange arrows.

Article Snippet: Similarly, on day 3, the cGMP ELISA kit (Bioss, China) was used to measure the amount of cGMP.

Techniques: Activation Assay, Membrane, Activity Assay

Journal: Bioactive Materials

Article Title: Shape memory bone screws loading L-arginine and Ca 2+ propagate mechanical stimulation, energize bone cells and augment bone regeneration

doi: 10.1016/j.bioactmat.2025.07.007

Figure Lengend Snippet: In vitro osteogenesis and angiogenesis potential of SMP bone screw. (A) Schematic illustration showing the released Ca 2+ and Arg synergistically activate the NO-cGMP pathway, ultimately promoting both osteogenesis and angiogenesis. Release profiles of (B) Ca 2+ and (C) Arg from different SMP bone screws. (D) Representative ALP staining images (with enlarged images inserted) and (E) quantitative analysis of ALP activity of rBMSCs on day 3 and day 7. (F) Representative ARS images and (G) quantitative analysis of calcium deposition of rBMSCs on day 7 and day 14. (H) Representative fluorescence images showing the endothelial network formation in HUVECs after co-culture for 6 and 12 h. (I-K) Relative expression of angiogenic-related gene expression including eNOS, nitrite levels, and cGMP concentration of rBMSCs and HUVECs on day 3. Data were presented as mean ± SD and analyzed by one-way ANOVA (n = 3 for each sample, ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001). These results suggest that Ca 2+ and Arg synergistically activate the NO-cGMP signaling pathway to promote osteogenesis and angiogenesis.

Article Snippet: Similarly, on day 3, the cGMP ELISA kit (Bioss, China) was used to measure the amount of cGMP.

Techniques: In Vitro, Staining, Activity Assay, Fluorescence, Co-Culture Assay, Expressing, Gene Expression, Concentration Assay

Journal: Bioactive Materials

Article Title: Shape memory bone screws loading L-arginine and Ca 2+ propagate mechanical stimulation, energize bone cells and augment bone regeneration

doi: 10.1016/j.bioactmat.2025.07.007

Figure Lengend Snippet: Bioinformatic analysis of gene expression of rMSCs on different bone screws. (A) Principal component analysis of gene expression profile of rMSCs grown in the presence of cSMP screw or SMP-Ca/Arg2-P screw; (B) Venn diagram illustration of the significant DEGs between the SMP-Ca/Arg2-P group and the cSMP group; (C) Volcano plots of transcriptomic analysis of DEGs in cSMP versus SMP-Ca/Arg2-P; (D) Up-regulated enriched Kyoto Encyclopedia of Genes and Genomes pathways of cSMP versus SMP-Ca/Arg2-P; (E) Heatmap evaluation of up-regulated DEGs involved in mechanosensitive calcium, PI3K-Akt and cGMP-PKG signaling pathways; (F-G) Relative messenger RNA expression evaluation of targeted genes via qRT-PCR; (H) Schematic illustration of potential SMP-Ca/Arg2-P-mediated activation of the mechanosensitive and NO-cGMP signaling pathway for coupling osteogenesis-angiogenesis. Sample size n = 3 for all experiments. Data were presented as mean ± SD and analyzed by one-way ANOVA ( n = 6 for each sample, ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001).

Article Snippet: Similarly, on day 3, the cGMP ELISA kit (Bioss, China) was used to measure the amount of cGMP.

Techniques: Gene Expression, Protein-Protein interactions, RNA Expression, Quantitative RT-PCR, Activation Assay