Journal: BioMedical Engineering OnLine

Article Title: Effects of immediate loading directionality on the mechanical sensing protein PIEZO1 expression and early-stage healing process of peri-implant bone

doi: 10.1186/s12938-024-01223-1

Figure Lengend Snippet: The comparison of peri-implant bone ECM and PIEZO1 expression between the compressive and tensile sides of peri-implant bone. a Peri-implant bone ECM was observed by VG staining in the compressive and tensile sides. Bone ECM (orange), the implant (black). Scale bar, 100 μm. b The bone ECM-to-implant contact (%) and the peri-implant bone ECM area (%) were quantified at ROIs in the compressive and tensile sides ( n = 9, Mann–Whitney test). c Immunofluorescence staining of PIEZO1 in the compressive and tensile sides. PIEZO1 (green), DAPI (blue). Scale bar, 500 μm. d Qualification of intensity and area fraction of positive PIEZO1 cells at ROIs in the compressive and tensile sides ( n = 9, Mann–Whitney test). All data are represented as mean ± SD. *** p < 0.001. NS no significance

Article Snippet: For bone ECM analysis, the sections were stained with VG (Sigma-Aldrich, USA) and visualized via the inverted microscope (Nikon eclipse, Japan).

Techniques: Comparison, Expressing, Staining, MANN-WHITNEY, Immunofluorescence

Journal: Bio-Design and Manufacturing

Article Title: Biofabrication of nanocomposite-based scaffolds containing human bone extracellular matrix for the differentiation of skeletal stem and progenitor cells

doi: 10.1007/s42242-023-00265-z

Figure Lengend Snippet: Schematic of the composite ink combinations used in this study

Article Snippet: Following 2 h stirring at room temperature, alginate (A, alginic acid sodium salt from brown algae, Sigma, UK) was added to the Laponite-bone-ECM (LB) suspension and homogenized with a spatula for 8–10 min to allow alginate inclusion.

Techniques:

Journal: Bio-Design and Manufacturing

Article Title: Biofabrication of nanocomposite-based scaffolds containing human bone extracellular matrix for the differentiation of skeletal stem and progenitor cells

doi: 10.1007/s42242-023-00265-z

Figure Lengend Snippet: Rheological properties of the nanoclay-based bone-ECM inks. a Viscosity over shear rate study of a series of nanoclay-based materials (a–i) in absence or (a–ii) inclusion of bone-ECM. b LAB gel over rheometer plates showing viscoelastic behavior. (c) Viscosity comparison at a fixed shear rate (10 s –1 ). d Storage and loss moduli of nanoclay-based materials (d–i, d–iii) without and (d–ii, d–iv) when blended with bone-ECM. Statistical significance was assessed by one-way ANOVA. Data are presented as mean±standard deviation, n =3, ∗∗∗∗ p <0.0001. ECM: extracellular matrix; LAB: Laponite-alginate-bone-ECM; ANOVA: analysis of variance

Article Snippet: Following 2 h stirring at room temperature, alginate (A, alginic acid sodium salt from brown algae, Sigma, UK) was added to the Laponite-bone-ECM (LB) suspension and homogenized with a spatula for 8–10 min to allow alginate inclusion.

Techniques: Viscosity, Shear, Comparison, Standard Deviation

Journal: Bio-Design and Manufacturing

Article Title: Biofabrication of nanocomposite-based scaffolds containing human bone extracellular matrix for the differentiation of skeletal stem and progenitor cells

doi: 10.1007/s42242-023-00265-z

Figure Lengend Snippet: Printing fidelity of nanocomposite bone-ECM inks. a Filament fusion test was carried out with (a–i) AB and (a–ii) LAB inks. b Measurements of the filament fusion tests performed with (b–i) 3% and (b–ii) 4% LAP composite inks. c Micrographs of scaffolds printed with (c–i) 3% and (c–ii) 4% LAP-based inks. Scale bar: 1 mm. AB: alginate-bone-ECM; LAB: Laponite-alginate-bone-ECM; LAP: Laponite; fs: fused segment length; ft: filament thickness; fd: filament distance

Article Snippet: Following 2 h stirring at room temperature, alginate (A, alginic acid sodium salt from brown algae, Sigma, UK) was added to the Laponite-bone-ECM (LB) suspension and homogenized with a spatula for 8–10 min to allow alginate inclusion.

Techniques:

Journal: Bio-Design and Manufacturing

Article Title: Biofabrication of nanocomposite-based scaffolds containing human bone extracellular matrix for the differentiation of skeletal stem and progenitor cells

doi: 10.1007/s42242-023-00265-z

Figure Lengend Snippet: HBMSC viability and proliferation post-printing. Live/dead assay was performed on 3D-printed a – c AB and d – f LAB scaffolds at Days 1, 7, and 21. g Cell viability and h density quantification following ImageJ analysis. j – m ALP staining of 3D bioprinted scaffolds following cultivation in basal (AB, j; LAB, l) and osteogenic (AB, k; LAB, m) media conditioning complete with acellular control (insets). n ALP intensity and o area coverage percentage. Scale bars: a – f 100 µm, j – m 50 µm (samples), 250 µm (acellular controls). Statistical significance was determined using two-way ANOVA. Data are presented as mean±standard deviation, n =3, ∗∗∗∗ p <0.0001. HBMSC: human bone marrow stromal cell; AB: alginate-bone-ECM; LAB: Laponite-alginate-bone-ECM; ALP: alkaline phosphatase; ANOVA: analysis of variance; O: osteogenic; B: basal

Article Snippet: Following 2 h stirring at room temperature, alginate (A, alginic acid sodium salt from brown algae, Sigma, UK) was added to the Laponite-bone-ECM (LB) suspension and homogenized with a spatula for 8–10 min to allow alginate inclusion.

Techniques: Live Dead Assay, Staining, Standard Deviation

Journal: Bio-Design and Manufacturing

Article Title: Biofabrication of nanocomposite-based scaffolds containing human bone extracellular matrix for the differentiation of skeletal stem and progenitor cells

doi: 10.1007/s42242-023-00265-z

Figure Lengend Snippet: Nanoclay-based inks support sustained release of VEGF in the CAM model. Macrographs during a sample implantation and b retrieval: (i) empty, (ii) AB, (iii) LAB, and VEGF-loaded (iv) AB and (v) LAB 3D-printed scaffolds. c Chalkley score of vascularized samples and controls. d – g Histological micrographs of samples stained for (i, ii) Goldner’s Trichrome and (iii, iv) Alcian Blue & Sirius Red. Statistical significance was assessed using one-way ANOVA. Data are presented as mean±standard deviation, n =4, * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001. Scale bars: a , b 10 mm, d – g 100 µm. VEGF: vascular endothelial growth factor; CAM: chick chorioallantoic membrane; AB: alginate-bone-ECM; LAB: Laponite-alginate-bone-ECM; ANOVA: analysis of variance

Article Snippet: Following 2 h stirring at room temperature, alginate (A, alginic acid sodium salt from brown algae, Sigma, UK) was added to the Laponite-bone-ECM (LB) suspension and homogenized with a spatula for 8–10 min to allow alginate inclusion.

Techniques: Staining, Standard Deviation, Membrane

Journal: Bio-Design and Manufacturing

Article Title: Biofabrication of nanocomposite-based scaffolds containing human bone extracellular matrix for the differentiation of skeletal stem and progenitor cells

doi: 10.1007/s42242-023-00265-z

Figure Lengend Snippet: Nanocomposite bone-ECM scaffolds support mineralization ex vivo. a Macro- and micro-graphs of empty control. Implanted and explanted b LAP-free and c LAP-loaded 3D (i) material (drug- and cell-free) control, (ii) BMP-2 loaded, (iii) cell-loaded, and (iv) BMP-2 and cell loaded scaffolds. d Chalkley score of implanted samples and control after 7 d of culture. e Quantitative analysis of afferent vascular supply to implanted scaffolds before extraction. f Micro-CT analysis of implanted scaffolds following 7 d of incubation in a CAM model. Scale bars: a – c 10 mm. Statistical significance was assessed by one-way ANOVA. Data are presented as mean±standard deviation, n =4, * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001. LAP: Laponite; 3D: three-dimensional; BMP-2: bone morphogenetic protein-2; micro-CT: microcomputed tomography; CAM: chick chorioallantoic membrane; ANOVA: analysis of variance; HBMSCs: human bone marrow stromal cells

Article Snippet: Following 2 h stirring at room temperature, alginate (A, alginic acid sodium salt from brown algae, Sigma, UK) was added to the Laponite-bone-ECM (LB) suspension and homogenized with a spatula for 8–10 min to allow alginate inclusion.

Techniques: Ex Vivo, Extraction, Micro-CT, Incubation, Standard Deviation, Tomography, Membrane

Journal: Bio-Design and Manufacturing

Article Title: Biofabrication of nanocomposite-based scaffolds containing human bone extracellular matrix for the differentiation of skeletal stem and progenitor cells

doi: 10.1007/s42242-023-00265-z

Figure Lengend Snippet: CAM implantation of 3D-printed scaffolds containing BMP-2 and HBMSCs. a – d LAP-free and e – h LAP-loaded groups are stained for (i, ii) Goldner’s Trichrome, (iii, iv) Alcian Blue & Sirius Red, and (v, vi) von Kossa. Scale bars: 100 µm. CAM: chick chorioallantoic membrane; 3D: three-dimensional; BMP-2: bone morphogenetic protein-2; HBMSCs: human bone marrow stromal cells; LAP: Laponite

Article Snippet: Following 2 h stirring at room temperature, alginate (A, alginic acid sodium salt from brown algae, Sigma, UK) was added to the Laponite-bone-ECM (LB) suspension and homogenized with a spatula for 8–10 min to allow alginate inclusion.

Techniques: Staining, Membrane