rat anti osteocalcin antibody  (TaKaRa)

Journal: Bioactive Materials

Article Title: Dual-function thermoresponsive antibiotic-loaded hydrogel with antimicrobial and osteogenic properties for implant-related infection control

doi: 10.1016/j.bioactmat.2026.02.044

Figure Lengend Snippet: In vivo immunohistochemical analysis (n = 6). (A) Representative immunolabeling images of the Control, PNVCL, and PNVCL/TC 25 mg/mL groups showing osteopontin (OPN) and osteocalcin (OCN) expression (immunopositive cells indicated by black arrows). (B) Mean scores (0–3) ± standard deviation for OPN immunostaining. (C) Mean scores (0–3) ± standard deviation for OCN immunostaining. Bars indicate statistically significant differences between groups (p < 0.05; Tukey HSD test).

Article Snippet: Endogenous peroxidase activity was quenched by incubation with 3% hydrogen peroxide for 1 h, followed by blocking of nonspecific binding sites with 1% bovine serum albumin for 12 h. The sections were then incubated with goat anti-osteopontin and goat anti-osteocalcin primary antibodies (sc-21742 and sc-30044, respectively; Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: In Vivo, Immunohistochemical staining, Immunolabeling, Control, Expressing, Standard Deviation, Immunostaining

Journal: Bioactive Materials

Article Title: MSC-mimicking nanovesicle embedded bio-adhesive hydrogel for dual immunomodulation and osteogenesis to promote maxillofacial bone regeneration

doi: 10.1016/j.bioactmat.2026.02.032

Figure Lengend Snippet: In vivo evaluation of bone regeneration after hydrogel implantation in the mandibular bone defect mouse model. a) Schematic illustration of the mandibular bone defect mouse model. b) Micro-CT 3D reconstruction images of the mandibular bone samples at 4 and 8 weeks post-surgery. Scale bar = 1 mm. c) Semi-quantitative analysis of BV/TV, bone surface, Tb.N and Tb.sp (n = 6) in mouse mandibular bone defects implanted with different hydrogels at 8 weeks post-surgery. d) H&E staining and Masson trichrome staining of tissue sections of mandibular defects at 8 weeks post-surgery. Scale bar = 100 μm. e, f) Immunofluorescent staining images and corresponding semi-quantitative analysis of the expression levels of RUNX2 and OCN in mandibular bone defect areas at 4 and 8 weeks post-surgery (n = 3). Scale bar = 50 μm. P-values are calculated using one-way ANOVA with Tukey's test, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, n.s. not significant ( a was created with bioRender. com).

Article Snippet: Briefly, tissue sections underwent antigen retrieval (37 °C, 30 min), blocked by 5% bovine serum albumin (BSA, RT, 1 h), then sequentially incubated (4 °C) with lineage-specific probes: macrophage marker F4/80 (1:200, Cat. sc-52664, Santa Cruz Biotechnology) and CD68 (1:250, Cat. 14-0681-81, ThermoFisher, USA), CD206 (1:500, Cat. 24595T, Cell Signaling Technology, USA) and Arg-1 (1:250, Cat. 82975, Proteintech, China) for a M2 marker, iNOS (1:500, Cat. ab178945, Abcam, UK) and CD86 (1:300, Cat. DF6332, Affinity, China) for a M1 marker, RUNX2 (1:150, Cat. sc390351, Santa Cruz Biotechnology, USA) and OCN (1:150, Cat. sc390877, Santa Cruz Biotechnology, USA) for osteogenesis markers, VEGF (1:50, Cat. sc57496, Santa Cruz Biotechnology, USA) and CD31 (1:50, Cat. sc20071, Santa Cruz Biotechnology, USA) for angiogenesis markers, and CD146 (1:200, Cat. Ab75769, Abcam, UK) for stem cell surface marker, overnight at 4 °C.

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

Journal: Bioactive Materials

Article Title: MSC-mimicking nanovesicle embedded bio-adhesive hydrogel for dual immunomodulation and osteogenesis to promote maxillofacial bone regeneration

doi: 10.1016/j.bioactmat.2026.02.032

Figure Lengend Snippet: The PEG-pp@nMSC@MT hydrogel effectively promotes BMMSCs' osteogenesis in vitro . a) Schematic illustration of co-culture BMMSCs with hydrogels. b) ALP staining of BMMSCs co-cultured with different scaffolds after 7 days. Scale bar = 500 μm. c) Semi-quantitative analysis of ALP staining (n = 3). d) ARS staining of BMMSC co-cultured with different scaffolds after 21 days. Scale bar = 500 μm. e) Semi-quantitative analysis of ARS staining (n = 3). f) mRNA expression of osteogenic genes (BMP2, OCN, and RUNX2) of BMMSCs treated with different hydrogels in the MMP condition after 5 and 10 days (n = 3). g) Western blot analysis of osteogenic protein (BMP2 and RUNX2) of BMMSCs treated with different hydrogels in the MMP condition after 7 and 14 days. P-values are calculated using one-way ANOVA with Tukey's test, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, n.s. not significant ( a was created with bioRender. com).

Article Snippet: Briefly, tissue sections underwent antigen retrieval (37 °C, 30 min), blocked by 5% bovine serum albumin (BSA, RT, 1 h), then sequentially incubated (4 °C) with lineage-specific probes: macrophage marker F4/80 (1:200, Cat. sc-52664, Santa Cruz Biotechnology) and CD68 (1:250, Cat. 14-0681-81, ThermoFisher, USA), CD206 (1:500, Cat. 24595T, Cell Signaling Technology, USA) and Arg-1 (1:250, Cat. 82975, Proteintech, China) for a M2 marker, iNOS (1:500, Cat. ab178945, Abcam, UK) and CD86 (1:300, Cat. DF6332, Affinity, China) for a M1 marker, RUNX2 (1:150, Cat. sc390351, Santa Cruz Biotechnology, USA) and OCN (1:150, Cat. sc390877, Santa Cruz Biotechnology, USA) for osteogenesis markers, VEGF (1:50, Cat. sc57496, Santa Cruz Biotechnology, USA) and CD31 (1:50, Cat. sc20071, Santa Cruz Biotechnology, USA) for angiogenesis markers, and CD146 (1:200, Cat. Ab75769, Abcam, UK) for stem cell surface marker, overnight at 4 °C.

Techniques: In Vitro, Co-Culture Assay, Staining, Cell Culture, Expressing, Western Blot

Journal: Bioactive Materials

Article Title: A continuous adhesion-enhanced osteogenic pathway in artificial scaffold drives cellular infiltration and condensed mineralization for rapid bone regeneration

doi: 10.1016/j.bioactmat.2026.02.026

Figure Lengend Snippet: In vitro study of osteogenic capacity and mechanisms of the CPH/rGO-3/0.6 scaffold (a) Fluorescent staining of hMSCs grown on the surface of Blank, CPH/rGO-3/0 and CPH/rGO-3/0.6 scaffolds for 7, 14 and 21 days and intensity statistics of osteocalcin (OCN) on 21 days (Cell nuclei of hMSCs were visualized using DAPI (blue); Cytoskeleton was stained with Phalloidin-FITC (green); OCN proteins were stained with Alexa Fluor 594 (red)) (n = 16, 12, 15 for Blank, CPH/rGO-3/0 and CPH/rGO-3/0.6 groups respectively. Data are expressed as mean ± SD. ∗ for p < 0.05; ∗∗ for p < 0.01; ∗∗∗ for p < 0.001). (b) Fluorescent staining of MSCs grown on the surface of CPH/rGO-3/0.6 scaffold for 28 days. (c) Osteogenesis related genes expression of MSCs including alkaline phosphatase ( ALP ), type I collagen (COL-I), runt-related transcription factor 2 ( Runx2 ), SP7 transcription factor ( SP7 ), Bone sialoprotein ( BSP ), dentin matrix acidic phosphoprotein 1( DMP1 ), OCN and osteopontin ( OPN ) after 7, 14 and 21 days' incubation on CPH/rGO-3/0, CPH/rGO-3/0.6 scaffolds and Blank (n = 3 per group. Data are expressed as mean ± SD. ∗ for p < 0.05; ∗∗ for p < 0.01; ∗∗∗ for p < 0.001). (d) OD value obtained from the ALP reagent of sample Blank, CPH/rGO-3/0 and CPH/rGO-3/0.6 scaffolds after osteogenic induction of hMSC for 4, 8 and 12 days (n = 3 per group. Data are expressed as mean ± SD. ∗ for p < 0.05; ∗∗ for p < 0.01; ∗∗∗ for p < 0.001). (e) Volcano map and (f) GO enrichment analysis of differentially expressed genes in hMSCs cultured on rGO/CS vs rGO and on CPH/rGO-3/0.6 vs CPH/rGO-3/0. (g) Hotmap of differentially expressed genes between rGO/CS and rGO samples, CPH/rGO-3/0.6 and CPH/rGO-3/0 scaffolds. (h) Western blot images of KCNN3 , Integrin β1 , ANK3 , FAK , MAPK , OCN , and BSP following 14 days of osteogenic induction co-culture of hMSCs with rGO, rGO/CS, Blank. (i) Schematic diagram of osteogenic gene pathways mediated by CPH/rGO-3/0.6.

Article Snippet: The staining of OCN was performed with polyclonal antibody to osteocalcin (Proteintech, USA) and goat anti-rabbit IgG H&L (Abcam, USA) after the blocking of bovine serum albumin (BSA, BioFroxx, Germany).

Techniques: In Vitro, Staining, Expressing, Incubation, Cell Culture, Western Blot, Co-Culture Assay

Journal: Bioactive Materials

Article Title: Ribbon-shaped microgels as bioinks for 3D bioprinting of anisotropic tissue structures

doi: 10.1016/j.bioactmat.2025.12.040

Figure Lengend Snippet: μRB bioinks modulate MSC morphology, osteogenesis and bone formation in a stiffness-dependent manner. (A) Live/dead staining of MSCs after extrusion (Day 0) or after 28 days of culture in osteogenic medium. Green: live cells, Red: dead cells. (B) Metabolic activity of MSCs measured by PrestoBlue assay at day 0 and day 14 after bioprinting (n = 6 per group). (C) DNA content per scaffold measured by PicoGreen assay at day 28 (n = 3 per group). (D) Alizarin red S (ARS) staining for mineralized bone matrix, (E) Aniline Blue staining for total collagen, and (F) immunostaining of Osteocalcin (OCN), a mature bone marker, at day 14 and day 21 of osteogenesis (n = 4 per group). (G–I) Quantification of ARS and Aniline Blue percent positive area and OCN mean fluorescence intensity (MFI). Scale bar = 100 μm. Values are presented as mean ± S.D. and p-values were determined by one-way analysis of variance (ANOVA) with Tukey's multiple comparisons test; ∗p ≤ 0.05, ∗∗p ≤ 0.01, ∗∗∗p ≤ 0.005, ∗∗∗∗p ≤ 0.001.

Article Snippet: Primary antibody for osteocalcin (1:200, 23418-1-AP, Proteintech), GFP (1:100, 50430-2-AP, Proteintech) was diluted in 1 % BSA with 0.1 % Triton X-100 and incubated overnight at 4 °C.

Techniques: Staining, Activity Assay, Prestoblue Assay, Picogreen Assay, Immunostaining, Marker, Fluorescence