trap staining instructions  (Beijing Solarbio Science)

Journal: Bioactive Materials

Article Title: Adaptive hydrogel loaded with pre-coordinated stem cells for enhanced osteoarthritis therapy

doi: 10.1016/j.bioactmat.2025.05.018

Figure Lengend Snippet: In vivo anti-inflammatory effects and inhibition of resorption in subchondral bone. (A and B) Immunofluorescence staining for the iNOS and Arg-1. (C) Micro-CT reconstructed images of subchondral bone tissues at week 4 with various treatments in OA modeled rat joints. (D) TRAP staining for osteoclasts. (E) The proportion of TRAP-stained positive cells. (F and G) Semiquantitative analysis of immunofluorescence staining for iNOS and Arg-1. Results are presented as mean ± SD (n ≥ 4). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: The procedure for staining is as described in the TRAP staining instructions (Solarbio, China).

Techniques: In Vivo, Inhibition, Immunofluorescence, Staining, Micro-CT

Journal: Bioactive Materials

Article Title: Adaptive hydrogel loaded with pre-coordinated stem cells for enhanced osteoarthritis therapy

doi: 10.1016/j.bioactmat.2025.05.018

Figure Lengend Snippet: Mg 2+ and DMOG synergize to inhibit osteoclast activation and modulate the cross-talk between chondrocytes and osteoclasts. (A) CLSM images showing F-actin in bone marrow mononuclear cells (BMMCs) after treatment with osteoclastic medium supplemented with Mg 2+ , DMOG, and M + D for 5 days, respectively. (B) Quantitative assay of TRAP activity at days 3 and 5. (C) TRAP staining of BMMCs. (D to F) Relative expression of mRNA (TRAP, MMP9, and Cathepsin K) of BMMCs. (G) SEM images showing the resorption lacunae formed on bovine dental slices at day 5 with different treatments. (H) Bone resorption area calculated based on G using ImageJ software. (I) Schematic illustration of the suppression of BMMCs differentiation into osteoclasts by Mg 2+ and DMOG. (J) Schematic representation of the interaction between osteoclasts and chondrocytes in Transwell culture. (K) TRAP staining of osteoclasts in the upper chambers. (L) Quantification of TRAP activity expressed by osteoclasts. (M) Analysis of the number of osteoclasts based on the staining of TRAP in K. (N) Staining for ALP, COL-X, and COL-II in chondrocytes located in the lower chambers. (O and P) Based on the relative fluorescence intensity distribution of COL-X and COL-II for the single cell in N. Results are presented as mean ± SD (n ≥ 4). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Article Snippet: The procedure for staining is as described in the TRAP staining instructions (Solarbio, China).

Techniques: Activation Assay, Activity Assay, Staining, Expressing, Software, Fluorescence

Journal: Materials Today Bio

Article Title: Self-powered intracellular nanogenerator attenuates inflammatory osteolysis through mitochondrial MRS2/Mg 2+ -mediated macrophage repolarization and osteoclastogenesis inhibition

doi: 10.1016/j.mtbio.2025.102050

Figure Lengend Snippet: pBTO inhibited osteoclastogenesis in vitro. (A) TRAP staining in RANKL-treated RAW264.7 cells stimulated with BTO and pBTO. (B) Relative mRNA levels of genes associated with osteoclastogenesis of RAW264.7 cells in the presence of RANKL, BTO or pBTO. (C) JC-1 staining in RANKL-treated RAW264.7 cells stimulated with BTO and pBTO. (D) Schematic diagram of pBTO inhibit osteoclastogenesis of macrophage (∗: p < 0.05, ∗∗: p < 0.01, ∗∗∗∗: p < 0.001).

Article Snippet: Based on our previous research [ ], RAW 264.7 cells were incubated with 100 μg/mL BTO and pBTO, and stimulated with 50 ng/mL RANKL (peprotech, USA) for 3–5 days and then subjected to TRAP staining (Servicebio, China).

Techniques: In Vitro, Staining

Journal: Materials Today Bio

Article Title: Self-powered intracellular nanogenerator attenuates inflammatory osteolysis through mitochondrial MRS2/Mg 2+ -mediated macrophage repolarization and osteoclastogenesis inhibition

doi: 10.1016/j.mtbio.2025.102050

Figure Lengend Snippet: pBTO improved inflammatory bone loss by modulating macrophage polarization and osteoclastogenesis in vivo. (A) Designed an mice inflammatory bone loss model. (B) Micro-CT scanning and 3D reconstruction of mice calvarial bone. pBTO inhibited LPS-induced inflammatory bone loss. (C) CD68 + Arg-1 + macrophages (arrow) in calvarial bone matrix under confocal microscope. PBTO increased the number of M2 macrophages. (D) CD68 + iNOS + macrophages (arrow) in calvarial bone matrix under confocal microscope. pBTO decreased the number of LPS-induced M1 macrophages. (E) HE staining of mice calvarial bone showed pBTO decreased LPS-induced inflammatory cell infiltration and bone resorption. (F) TRAP staining of mice calvarial bone showed pBTO decreased the number of osteoclasts.

Article Snippet: Based on our previous research [ ], RAW 264.7 cells were incubated with 100 μg/mL BTO and pBTO, and stimulated with 50 ng/mL RANKL (peprotech, USA) for 3–5 days and then subjected to TRAP staining (Servicebio, China).

Techniques: In Vivo, Micro-CT, Microscopy, Staining