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

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