osteogenic medium  (ATCC)

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: Journal of Translational Medicine

Article Title: Engineering a vascularized-osteogenic microenvironment to enhance bone regeneration via a 3D-printed composite scaffold with progressive-release bio-factors

doi: 10.1186/s12967-026-08090-5

Figure Lengend Snippet: Characterization of biocompatibility and osteogenic inductive capacity of scaffold materials. (a) Cytotoxicity assessment of PCL, PHA, and PHL scaffolds against bone marrow mesenchymal stem cells (BMSCs) via Live/Dead staining. Green: Calcein-AM (live cells), Red: Propidium iodide (PI, apoptotic cells) (Scale bar = 200 μm). (b) Proliferation and viability of BMSCs co-cultured with scaffolds for 1, 3, and 5 days, determined by CCK-8 assay. (c) Representative images of scratch wound healing assay (Scale bar = 200 μm). (d) Quantitative analysis of cell migration rate. (e) Alkaline phosphatase (ALP) staining of BMSCs after osteogenic differentiation induction (Scale bar = 200 μm). (f) ALP staining of BMSCs directly co-cultured with scaffolds without osteogenic supplements(Scale bar=1 mm). (g) Alizarin red S (ARS) staining of BMSCs after osteogenic differentiation induction (Scale bar = 200 μm). (h) ARS staining of BMSCs directly co-cultured with scaffolds without osteogenic supplements(Scale bar=1 mm). (i) Quantitative analysis of ALP-positive staining area. (j) Quantitative analysis of ARS-positive mineralized nodule area. All experimental data are expressed as mean ± SD ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: DMEM medium, osteogenic differentiation medium, the Cell Counting Kit-8, and the Live/Dead Cell Kit were purchased from Solarbio.

Techniques: Staining, Cell Culture, CCK-8 Assay, Wound Healing Assay, Migration

Journal: Journal of Translational Medicine

Article Title: Engineering a vascularized-osteogenic microenvironment to enhance bone regeneration via a 3D-printed composite scaffold with progressive-release bio-factors

doi: 10.1186/s12967-026-08090-5

Figure Lengend Snippet: Osteogenic promotion of GV@PHL scaffold in vivo. ( a ) Schematic illustration of critical-size bone defect modeling in SD rats and GV@PHL transplantation. (b) Macroscopic images of rat calvarial bone defects treated with different scaffolds at 6 weeks. (c) Bone defect traces and defect rates of each experimental group after various treatments. (d) Micro-CT 3D reconstruction images of rat calvarial bone defects treated with different scaffolds at 6 weeks post-surgery. e-h) Bone mineral density (BMD), bone volume/tissue volume (BV/TV), bone surface/total volume (BS/TV), and trabecular number (Tb.N) of bone defects treated with different scaffolds. i-j) HE staining and Masson trichrome staining results of each group (black scale bar = 1 mm, red scale bar = 50 μm). All experimental data are expressed as mean ± SD ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: DMEM medium, osteogenic differentiation medium, the Cell Counting Kit-8, and the Live/Dead Cell Kit were purchased from Solarbio.

Techniques: In Vivo, Transplantation Assay, Micro-CT, Staining

Journal: Dose-Response

Article Title: Skeletal Stem Cells Rescue Radiation-Induced Osteogenic Precursor Cell Dysfunction via the Wnt/β-Catenin Signaling Pathway

doi: 10.1177/15593258261440983

Figure Lengend Snippet: Co-culture with SSCs rescues the function of irradiated osteogenic precursor cells. (A, B) Cell apoptosis was analyzed by flow cytometry with Annexin V-PE/7AAD double staining. (A) Representative flow cytometry plots. (B) Quantitative analysis of the apoptotic rate. (C, D) ALP activity was assessed. (C) Representative ALP staining images (Scale bar: 50 μm). (D) Quantitative analysis of the relative ALP activity. (E, F) Mineralization capacity was evaluated using Alizarin Red S staining. (E) Representative staining images of mineralized nodules (Scale bar: 100 μm). (F) Quantitative analysis of the relative mineralization level. (G, H) Cell migration was determined by a migration assay. (G) Representative images of migrated cells (Scale bar: 50 μm). (H) Quantitative analysis of the relative cell migration level. All data are presented as mean ± SD, with statistical significance determined by unpaired two-tailed Student’s t-test (* p < 0.05; ** p < 0.01).

Article Snippet: After irradiation and corresponding interventions, cells were cultured in osteogenic induction medium (iXCells Biotechnologies, San Diego, CA, Cat. No. MD-0006) for 7 days.

Techniques: Co-Culture Assay, Irradiation, Flow Cytometry, Double Staining, Activity Assay, Staining, Migration, Two Tailed Test

Journal: Dose-Response

Article Title: Skeletal Stem Cells Rescue Radiation-Induced Osteogenic Precursor Cell Dysfunction via the Wnt/β-Catenin Signaling Pathway

doi: 10.1177/15593258261440983

Figure Lengend Snippet: SSCs exert rescue effects via the Wnt/β-catenin signaling pathway. (A) Representative ALP staining images of cells in each group (Scale bar: 50 μm). (B) Quantitative analysis of ALP activity in each group. (C) Representative Alizarin Red S staining images of cells in each group (Scale bar: 100 μm). (D) Quantitative analysis of Alizarin Red S staining in each group. (E) Relative mRNA expression levels of osteogenic marker genes ( Runx2 , Col1a1 , and OCN ) detected by qRT-PCR. GAPDH was used as an internal reference gene. (F) Representative Western blot images showing the expression levels of RUNX2, COL1A1, OCN, and β-catenin in each group. GAPDH was used as a loading control. (G) Quantitative analysis of Western blot results (gray value ratio of target protein to GAPDH) in each group. All data are presented as mean ± SD, with statistical significance determined by unpaired two-tailed Student’s t-test (* p < 0.05; ** p < 0.01; *** p < 0.001).

Article Snippet: After irradiation and corresponding interventions, cells were cultured in osteogenic induction medium (iXCells Biotechnologies, San Diego, CA, Cat. No. MD-0006) for 7 days.

Techniques: Staining, Activity Assay, Expressing, Marker, Quantitative RT-PCR, Western Blot, Control, Two Tailed Test

Journal: Dose-Response

Article Title: Skeletal Stem Cells Rescue Radiation-Induced Osteogenic Precursor Cell Dysfunction via the Wnt/β-Catenin Signaling Pathway

doi: 10.1177/15593258261440983

Figure Lengend Snippet: SSCs alleviate the radiation-induced bone injury in mice. (A–G) Micro-CT analysis of bone microstructure. (A) Representative micro-CT images of femurs. Quantitative analysis of (B) bone mineral density (BMD), (C) bone volume fraction (BV/TV), (D) trabecular thickness (Tb.Th), (E) trabecular number (Tb.N), (F) connectivity density (Conn.D), and (G) trabecular separation (Tb.Sp) at 2- and 4-weeks post irradiation. (H–K) Histological analysis (Scale bar: 100 μm). (H) H&E staining showing steatosis (arrows) and (I) quantitative analysis of steatotic lesions per field. (J) TRAP staining showing osteoclasts (arrows) and (K) quantitative analysis of osteoclast number per field. (L–O) Immunohistochemical staining of osteogenic markers (Scale bar: 100 μm). (L) Osterix staining and (M) quantitative analysis of Osterix-positive area. (N) β-catenin staining and (O) quantitative analysis of β-catenin-positive area. All experiments were conducted in three groups: Control, irradiation (IR), and IR plus SSC (IR+SSC) at 2- and 4-weeks post-irradiation. All data are presented as mean ± SD, with statistical significance determined by unpaired two-tailed Student’s t-test (* p < 0.05; ** p < 0.01; *** p < 0.001)

Article Snippet: After irradiation and corresponding interventions, cells were cultured in osteogenic induction medium (iXCells Biotechnologies, San Diego, CA, Cat. No. MD-0006) for 7 days.

Techniques: Micro-CT, Irradiation, Staining, Immunohistochemical staining, Control, Two Tailed Test