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Figures
- Fig. 1.
3D printed hydroxyapatite scaffold with defined macroporosity. Scale bars: 0.5 cm (A), 500 µm (B) and 5 µm (C).
- Fig. 2.
Cell attachment and proliferation on 3D-printed HA scaffold under static versus dynamic conditions. (A) MTT staining after 18 hours of cell seeding under static (left) and dynamic (right) conditions. (B) Cell number based on DNA content of cells seeded on untreated or oxygen-plasma treated (OPT) scaffolds for up to 28 days of culture under static or dynamic conditions. (C) SEM images of MG-63 cells cultivated on 3D-printed HA scaffolds after 18 h, 7 and 28 days of culture in static (left column) and dynamic conditions (middle column) and of hBMSCs cultivated under dynamic conditions (right column), scale bar: 10 μm. Lower panels show histological H&E stained sections of corresponding cell-seeded scaffolds after 28 days (scale bars: 500 μm).
- Fig. 3.
Differentiation of MG-63 human osteoblast-like cells cultured on 3D-printed porous HA scaffolds under static versus dynamic conditions. Real-time PCR analysis of (A) ALP (left) and ALP activity (right); (B) Collagen I and (C) Osteocalcin expressed by MG-63 cells under static and dynamic conditions after 3, 7, 14 and 28 days of culture, and corresponding immunofluorescence staining after 28 days of culture under static and dynamic conditions. Scale bars: 20 µm.
- Fig. 4.
Real-time PCR analysis of bone-associated genes expressed by hBMSC cells cultured in dynamic condition on 3D-porous HA scaffolds. (A) ALP. (B) Collagen I. (C) Osteocalcin. Cultured in proliferation medium or differentiation medium (DI).
- Fig. 5.
Real-time PCR analysis angiogenesis-related genes expressed by hBMSC cells cultured under dynamic condition on 3D-porous HA scaffolds and typical angiogenesis assay. The CAM assay allows in ovo vascularistion of biomaterials planted on the surface. (A–C) Manifold gene expression of (A) VEGF, (B) CD31and (C) eNOS cultured in proliferation medium (grey bars) or osteogenic differentiation medium (DI, black bars). (D) Windowed egg at incubation day ID7 when HA scaffolds were onplanted (left) and vascularized HA scaffold coated with ECM from osteoinductive medium at ID14 after 1 week on the CAM (right). (E) Relative relaxation rates as assessed in the MRI of HA scaffolds decorated with ECM produced by cells cultivated in osteogenic medium with differentiation induction (black bars) or in proliferative medium (light grey bars) and of uncoated HA scaffolds (dark grey bars). For the precise composition of the two culture media, see Materials and Methods section. A scheme (on top) represents the different regions of the scaffold (surface, middle and interface) of an egg.
- Fig. 6.
Overview of experimental design. Steps to tailor the 3D-printed ceramic scaffold (left) and methods used, as well as the parameters assessed at each step (right).
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