Journal: Polymers

Article Title: 3D Printed Scaffold Based on Type I Collagen/PLGA_TGF-β1 Nanoparticles Mimicking the Growth Factor Footprint of Human Bone Tissue

doi: 10.3390/polym14050857

Figure Lengend Snippet: TGF-β1 staining in bone tissue and semi-quantitative analysis ( A – D ): TGF-β1 staining in cortical ( A , B ) and trabecular ( C , D ) bone tissue. (( A , C ) 10× magnification; ( B , D ) 20× magnification); black arrows highlight the protein accumulation in bone ECM. ( E ) Histogram representing area percentage of TGF-β1 staining.

Article Snippet: The experimental data for TGF-β1 expression in human bone ECM were analysed by GraphPad Prism 7 software ( https://www.graphpad.com/scientific-software/prism/ , accessed on 10 September 2018).

Techniques: Staining

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Decellularized vascularized bone grafts: A preliminary in vitro porcine model for bioengineered transplantable bone shafts

doi: 10.3389/fbioe.2022.1003861

Figure Lengend Snippet: Mechanical Testing bone ECM. (A–C) : Bending test performed on radial and ulnar shaft. Visualization of the fracture (A) of the rod after successful bending test. Radius (B) and ulna (C) 3-pt bending curves (red = native, blue = decellularized) achieved while plotting in a graph the force (N) versus vertical displacement (mm). (D–F) Hardness test on radial and ulnar bone segments. Visualization of the tip print on the mounted sample (D). Radius (E) and ulna (F) hardness measures in native (red) and decellularized bone ECM (blue), Hardness values (HV) were expressed in Vicker units ± SD (n = 3 each; **** p < 0.0001)

Article Snippet: Approximately 5 × 10 5 cells were seeded on each decellularized bone ECM under two conditions (n = 3 for each) including: 1) bone ECM + proliferation medium (PM = DMEM containing, 2 mM l -glutamine and supplemented with 1% P/S and 2.5 μg/ml of amphotericin-B 1 ml/ml); 2) bone ECM + differentiation medium (DM = PM + 1 mM dexamethasone (D4902, Sigma-Aldrich), 50 ng/ml of sodium ascorbate (A4034, VWR), and 36 mg/ml of sodium dihydrogen phosphate monohydrate (1.06346, Sigma-Aldrich)).

Techniques:

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Decellularized vascularized bone grafts: A preliminary in vitro porcine model for bioengineered transplantable bone shafts

doi: 10.3389/fbioe.2022.1003861

Figure Lengend Snippet: Cytocompatibility study of periosteum and bone ECM with fibroblastic cell line. (A–C) : Live/Dead staining of NIH3T3 seeded periosteum ECM (top) and bone ECM (bottom) discs after 3 (A) , 7 (B) and 16 (C) days of culture, respectively, at 2.5 x gross magnification. All scale bars = 1,000 µm. (D–E) : H&E staining of NIH-3T3 seeded periosteum ECM (top) and bone ECM (bottom) after 3 (D) , 7 (E) and 16 (F) days of culture, respectively, revealing adherent fibroblasts (black arrows) at 40x gross magnification. All scale bars = 50 µm. (G) Cell viability analyzed with four different Live/Dead images at ×2.5 gross magnification of seeded periosteum and culture control wells after seven culture days (n = 3 for each group; p = 0.7987, ns = non-significant). (H) Cell proliferation rate analyzed using Prestoblue Assay during 16 days of culture (n = 3 × 2 ECM; ns = non-significant).

Article Snippet: Approximately 5 × 10 5 cells were seeded on each decellularized bone ECM under two conditions (n = 3 for each) including: 1) bone ECM + proliferation medium (PM = DMEM containing, 2 mM l -glutamine and supplemented with 1% P/S and 2.5 μg/ml of amphotericin-B 1 ml/ml); 2) bone ECM + differentiation medium (DM = PM + 1 mM dexamethasone (D4902, Sigma-Aldrich), 50 ng/ml of sodium ascorbate (A4034, VWR), and 36 mg/ml of sodium dihydrogen phosphate monohydrate (1.06346, Sigma-Aldrich)).

Techniques: Staining, Prestoblue Assay

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Decellularized vascularized bone grafts: A preliminary in vitro porcine model for bioengineered transplantable bone shafts

doi: 10.3389/fbioe.2022.1003861

Figure Lengend Snippet: Recellularization of decellularized bone-ECM with pAMSC and osteogenic differentiation. (A–D) : Alizarin red staining performed directly in the control culture wells seeded with pAMSC after 7 days of culture with DM (A) or PM (B), as well as in the culture wells with pAMSC seeded on bone ECM cultured with DM (C) or PM (D) after 22 culture days showed the formation of calcified micro-nodules (*) in all conditions excepting the seeded cells cultured with PM only. All scale bars = 200 μm. (E–G) IHC staining of respectively GFP (E) , osteocalcin (F), and negative control (G) of seeded pAMSC on bone ECM cultured with either DM (left) or PM (right) at 22 days of culture. Black arrows showed the positive osteocalcin staining corresponding to the GFP-pAMSC (All scale bars = 100 µm).

Article Snippet: Approximately 5 × 10 5 cells were seeded on each decellularized bone ECM under two conditions (n = 3 for each) including: 1) bone ECM + proliferation medium (PM = DMEM containing, 2 mM l -glutamine and supplemented with 1% P/S and 2.5 μg/ml of amphotericin-B 1 ml/ml); 2) bone ECM + differentiation medium (DM = PM + 1 mM dexamethasone (D4902, Sigma-Aldrich), 50 ng/ml of sodium ascorbate (A4034, VWR), and 36 mg/ml of sodium dihydrogen phosphate monohydrate (1.06346, Sigma-Aldrich)).

Techniques: Staining, Cell Culture, Immunohistochemistry, Negative Control