Abstract
Bone tissue is highly vascularized. The crosstalk of vascular and osteogenic cells is not only responsible for the formation of the strongly divergent tissue types but also for their physiological maintenance and repair. Extrusion-based bioprinting presents a promising fabrication method for bone replacement. It allows for the production of large-volume constructs, which can be tailored to individual tissue defect geometries. In this study, we used the all-gelatin-based toolbox of methacryl-modified gelatin (GM), non-modified gelatin (G) and acetylated GM (GMA) to tailor both the properties of the bioink towards improved printability, and the properties of the crosslinked hydrogel towards enhanced support of vascular network formation by simple blending. The vasculogenic behavior of human dermal microvascular endothelial cells (HDMECs) and human adipose-derived stem cells (ASCs) was evaluated in the different hydrogel formulations for 14 days. Co-culture constructs including a vascular component and an osteogenic component (i.e. a bone bioink based on GM, hydroxyapatite and ASCs) were fabricated via extrusion-based bioprinting. Bioprinted co-culture constructs exhibited functional tissue-specific cells whose interplay positively affected the formation and maintenance of vascular-like structures. The setup further enabled the deposition of bone matrix associated proteins like collagen type I, fibronectin and alkaline phosphatase within the 30-day culture.
Highlights
By HDMECs and ASCs, and regarding their suitability for extrusion-based bioprinting
The optimized bioinks allowed the bioprinting of co-culture hydrogels including HDMECs and ASCs for the vascular compartment, and ASCs for the osteogenic compartment
Cellular crosstalk led to further differentiation of the ASCs in the osteogenic compartment, which was observed by expression of bone-specific proteins Col I, OPN and FN, and a self-guided, stabilized assembly of capillary-like networks in the vascular compartment
Summary
By HDMECs and ASCs, and regarding their suitability for extrusion-based bioprinting. The production of GM2 (in this study named GM) and GM2A8 (in this study named GMA) is described in[32,39], with the subscript numbers indicating the mode of methacrylation or acetylation, respectively. The average degrees of modification are 0.32 mmol g−1 methacryloyl per GM2, 0.38 mmol g−1 methacryloyl per GM2A8, 0.4 mmol g−1 acetyl per GM2A8. The following formulations of bioinks were prepared by solubilizing the biopolymers in phosphate buffered saline (PBS) at 60 °C for at least 60 mi
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