Journal: Bioactive Materials

Article Title: A motion-activated skeletal “Pacemaker” for systemic bone remodeling via oral piezoelectric microspheres

doi: 10.1016/j.bioactmat.2026.05.051

Figure Lengend Snippet: In vitro evaluation of the osteogenic-promoting capability of BTO@ALN. (A–D) Immunofluorescence staining of osteoblasts differentiated from primary mouse bone marrow–derived mesenchymal stem cells (BMSCs) (red: actin; green: RUNX2 and ATF4) and corresponding quantitative analysis of fluorescence intensity. (E–H) Alkaline phosphatase (ALP) and Alizarin Red S (ARS) staining of osteoblasts derived from primary mouse BMSCs and quantitative analysis of staining intensity.

Article Snippet: Osteogenic differentiation was induced by culturing BMSCs in osteogenic induction medium (Procell, Wuhan, China).

Techniques: In Vitro, Immunofluorescence, Staining, Derivative Assay, Fluorescence

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: Diagram of preparation and function design of bone tissue scaffolds. (a) Schematic illustration of the formation mechanism of the self-assembled process of porous CPH/rGO-3/0.6 (CS/PVA/HA/rGO) composite scaffolds with continuous graphene surface. (b) Mechanism of cell adhesion and migration on the surface of the CPH/rGO-3/0.6 scaffolds and (c) the osteogenic differentiation and biomineralization of MSCs on the modified rGO surface in the porous CPH/rGO-3/0.6 scaffolds. (d) Schematic illustration of the process of CPH/rGO-3/0.6 artificial bone implantation and the rapid ingrowth of new bone.

Article Snippet: The osteogenic induction medium was prepared with α-minimum essential medium (α-MEM, Gibco, USA) with 10 % fetal bovine serum (FBS, Gibco, USA), 1 % antimicrobial of penicillin, 8 nM β-Glycerol phosphate disodium (≧98 %, Solarbio, Beijing, China), 1 × 10 −8 dexamethasone (≧98 %, Solarbio, Beijing, China) and 2 × 10 −4 Vitamin C (≧ 99 %, Solarbio, Beijing, China), and osteogenic medium was changed every 2 days.

Techniques: Migration, Modification

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: Calcium deposition capacity of rGO/CS substrate and CPH/rGO-3/0.6 scaffold. (a) Crystallization on the surfaces of glass coverslip, rGO and rGO/CS. (b) Calcium nodules generated by hMSC on rGO and rGO/CS surfaces after 21 days of osteogenic induction. SEM images and EDS mapping of calcium nodules (c) on the surface of rGO/CS plate, (d) on the surface of hMSC and (e) in the hMSC cultured on the rGO/CS surface after 21 days of induction. (f) TEM images of calcium nodules generated by hMSCs on rGO and rGO/CS after 21 days of induction and the HRTEM image of calcium nodules generated by hMSCs and its SAED pattern. (g) SEM images of hMSCs on CPH/rGO-3/0 and CPH/rGO-3/0.6 scaffolds after osteogenic induction for 7, 14 and 21 days and corresponding content of element Ca on 21 days. (h) SEM images of calcium deposition of hMSC on CPH/rGO-3/0.6 scaffolds after osteogenic induction for 21 days and corresponding C, O, Ca and P elemental mapping. (i) SEM images of calcium deposition of hMSC on CPH/rGO-3/0.6 scaffolds after osteogenic induction for 28 days and its corresponding C, O, Ca and P elemental mapping.

Article Snippet: The osteogenic induction medium was prepared with α-minimum essential medium (α-MEM, Gibco, USA) with 10 % fetal bovine serum (FBS, Gibco, USA), 1 % antimicrobial of penicillin, 8 nM β-Glycerol phosphate disodium (≧98 %, Solarbio, Beijing, China), 1 × 10 −8 dexamethasone (≧98 %, Solarbio, Beijing, China) and 2 × 10 −4 Vitamin C (≧ 99 %, Solarbio, Beijing, China), and osteogenic medium was changed every 2 days.

Techniques: Crystallization Assay, Generated, Cell Culture

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 osteogenic induction medium was prepared with α-minimum essential medium (α-MEM, Gibco, USA) with 10 % fetal bovine serum (FBS, Gibco, USA), 1 % antimicrobial of penicillin, 8 nM β-Glycerol phosphate disodium (≧98 %, Solarbio, Beijing, China), 1 × 10 −8 dexamethasone (≧98 %, Solarbio, Beijing, China) and 2 × 10 −4 Vitamin C (≧ 99 %, Solarbio, Beijing, China), and osteogenic medium was changed every 2 days.

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

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: Regeneration of bone defects with critical size. (a) 3D images reconstructed with Micro-CT and X-ray images of blank, CPH/rGO-3/0, CPH/rGO-3, HA and 3D Printing scaffolds after implantation for 3 months. (b) Statistics of osteogenic parameters based on Micro-CT (n = 6 per group. Data are expressed as mean ± SD. ∗ for p < 0.05; ∗∗ for p < 0.01; ∗∗∗ for p < 0.001). H&E and Masson's staining of (c) entire defect area and (d) the junction between implanted scaffolds and native bone and inside of different scaffolds after implantation for 3 months. (e) Schematic illustrations of the ingrowth of new bone into different scaffolds. (f) H&E staining of CPH/rGO-3/0 and CPH/rGO-3/0.6 scaffold and their crystallization characterized through POM and TEM after implantation for 3 months. (g) SEM images and EDS mapping of the entire implant area and images at high magnification of interface between defect area (D) and natural bone (B), and inside of the scaffolds. (h) SEM images and EDS mapping of interface between CPH/rGO-3/0.6 scaffold and new bone on tissue section. (i) Three-point bending tests of different scaffolds in the femoral hemisection model after implantation for 1 month (n = 5 per group. Data are expressed as mean ± SD. ns, no statistical significance. ∗ for p < 0.05; ∗∗ for p < 0.01; ∗∗∗ for p < 0.001).

Article Snippet: The osteogenic induction medium was prepared with α-minimum essential medium (α-MEM, Gibco, USA) with 10 % fetal bovine serum (FBS, Gibco, USA), 1 % antimicrobial of penicillin, 8 nM β-Glycerol phosphate disodium (≧98 %, Solarbio, Beijing, China), 1 × 10 −8 dexamethasone (≧98 %, Solarbio, Beijing, China) and 2 × 10 −4 Vitamin C (≧ 99 %, Solarbio, Beijing, China), and osteogenic medium was changed every 2 days.

Techniques: Micro-CT, Staining, Crystallization Assay

Journal: Bioactive Materials

Article Title: Sulfated polysaccharide prevents senescent adipocyte-driven osteonecrosis by stem cell fate reprogramming

doi: 10.1016/j.bioactmat.2025.11.039

Figure Lengend Snippet: Continuous intraosseous administration of SCS prevents glucocorticoid-induced bone degeneration. ( A ) Schematic illustration of the glucocorticoid (GC; MPS)-induced bone deterioration and intraosseous SCS treatment. ( B-D ) Representative H&E staining images of the femur at 6 weeks (B). Magnified views of the cortical bone and trabecular bone in the marrow cavity are shown on the right. Solid arrows indicate normal osteocytes, while hollow arrows indicate empty osteocyte lacunae. Quantification of empty lacunae ratios in cortical bone (C) and trabecular bone (D). n = 6 biological replicates. (Scale bars, 500 μm and 25 μm) ( E-H ) Representative immunofluorescence staining of OPN + mature osteoblasts, osteolectin + osteoprogenitors, and VE-cadherin + endothelial cells (ECs) in femur at 6 weeks (E), and corresponding quantifications (F–H). n = 6 biological replicates. (Scale bars, 100 μm and 20 μm) ( I and J ) Representative flow cytometry plots of capillary subtypes in the femur (I), with quantification of CD45 − Ter119 − CD31 hi Emcn hi ECs (J). n = 6 biological replicates. ( K and L ) Flow cytometry plots showing Sca-1 hi CD31 hi arteriolar ECs (K), and corresponding quantification (L). n = 6 biological replicates. ( M and N ) Representative micro-CT 3D images of the femur (M). Quantitative analysis of percent bone volume (BV/TV) (N). n = 6 biological replicates. (Scale bars, 1.5 mm, 600 μm and 545 μm) ( O and P ) ELISA analysis of VEGF (O) and PDGF-BB (P) levels in bone marrow supernatant and peripheral serum from PBS- and SCS-treated groups at week 6. n = 6 biological replicates. ( Q ) ELISA quantification of the osteogenic factor osteocalcin in peripheral serum at week 6. n = 6 biological replicates. Data are presented as mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001; ns, not significant. Statistical significance was determined using one-way ANOVA with Tukey's post hoc test ( C, D, F, G, H, J, L, N, O, P and Q ).

Article Snippet: Serum concentrations of the osteogenic marker osteocalcin (NOVUS, NBP2-68151) were also measured.

Techniques: Staining, Immunofluorescence, Flow Cytometry, Micro-CT, Enzyme-linked Immunosorbent Assay

Journal: Bioactive Materials

Article Title: Sulfated polysaccharide prevents senescent adipocyte-driven osteonecrosis by stem cell fate reprogramming

doi: 10.1016/j.bioactmat.2025.11.039

Figure Lengend Snippet: SCS targets downstream senescent lineage commitment of bone marrow MSCs to mitigate GC-induced bone deterioration. ( A ) Schematic diagram illustrating the experimental design: CD45 − Ter119 − CD31 − LepR + MSCs isolated from mice co-treated with SCS and MPS for 7 days were subjected to in vitro lineage-competitive differentiation, followed by DEX-induced senescence in lineage-mixed cells. These cells were then adoptively transplanted into healthy bone marrow cavity to assess bone deterioration development. ( B ) Representative H&E-stained images of the femur 12 weeks after adoptive transfer. PBS-DEX group: LepR + MSCs from PBS and MPS co-treated mice subjected to in vitro lineage differentiation and DEX-induced senescence, followed by transplantation. SCS-DEX group: LepR + MSCs from SCS and MPS co-treated mice processed similarly. PBS group: solvent control without cell transplantation. Solid arrows indicate intact osteocytes; hollow arrows indicate empty lacunae. (Scale bars, 250 μm and 25 μm) ( C – E ) Quantitative analysis of marrow hypertrophic adipocyte diameter (C), proportion of empty osteocyte lacunae in trabecular bone (D), and adipocyte number (E) in the metaphysis 12 weeks post-transplantation. n = 19 biological replicates (C), n = 6 biological replicates (D), n = 8 biological replicates (E). ( F ) Quantification of empty lacunae in epiphysis at 12 weeks post-transplantation. n = 6 biological replicates. ( G – I ) Representative flow cytometry plots of capillary ECs subtypes in the femur at 12 weeks (G), with quantification of CD45 − Ter119 − CD31 hi Emcn hi ECs (H) and CD45 − Ter119 − CD31 lo Emcn lo ECs (I). n = 6 biological replicates. ( J and K ) Representative flow cytometry plots (J) and corresponding quantification (K) of CD45 − Ter119 − Sca-1 hi CD31 hi arteriolar ECs in the femur at 12 weeks post-transplantation. n = 6 biological replicates. ( L ) Representative micro-CT images of the femur at 12 weeks post-transplantation across different treatment groups. (Scale bars, 1.5 mm and 500 μm) ( M – P ) Quantitative analysis of bone parameters in the metaphysis: bone mineral density (BMD) (M), percent bone volume (BV/TV) (N), trabecular separation (Tb.Sp) (O), and trabecular number (Tb.N) (P). n = 6 biological replicates. ( Q ) Serum ELISA analysis of the osteogenic marker osteocalcin at 12 weeks post-transplantation. n = 6 biological replicates. ( R and S ) ELISA analysis of PDGF-BB (R) and VEGF (S) in both bone marrow supernatant and peripheral serum at 12 weeks post-transplantation. n = 6 biological replicates. Data are presented as mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001; ns, not significant. Statistical significance was determined using one-way ANOVA with Tukey's post hoc test ( C, D, E, F, H, I, K, M, N, O, P, Q, R and S ).

Article Snippet: Serum concentrations of the osteogenic marker osteocalcin (NOVUS, NBP2-68151) were also measured.

Techniques: Isolation, In Vitro, Staining, Adoptive Transfer Assay, Transplantation Assay, Solvent, Control, Flow Cytometry, Micro-CT, Enzyme-linked Immunosorbent Assay, Marker