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Engineering microvasculature by 3D bioprinting of prevascularized spheroids in photo-crosslinkable gelatin

Lise De Moor (UGent) , Jasper Smet, Magaly Plovyt (UGent) , Bieke Bekaert (UGent) , Chris Vercruysse (UGent) , Mahtab Asadian (UGent) , Nathalie De Geyter (UGent) , Sandra Van Vlierberghe (UGent) , Peter Dubruel (UGent) and Heidi Declercq (UGent)
(2021) BIOFABRICATION. 13(4).
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Abstract
To engineer tissues with clinically relevant dimensions by three-dimensional bioprinting, an extended vascular network with diameters ranging from the macro- to micro-scale needs to be integrated. Extrusion-based bioprinting is the most commonly applied bioprinting technique but due to the limited resolution of conventional bioprinters, the establishment of a microvascular network for the transfer of oxygen, nutrients and metabolic waste products remains challenging. To answer this need, this study assessed the potential and processability of spheroids, containing a capillary-like network, to be used as micron-sized prevascularized units for incorporation throughout the bioprinted construct. Prevascularized spheroids were generated by combining endothelial cells with fibroblasts and adipose tissue-derived mesenchymal stem cells as supporting cells. To serve as a viscous medium for the bioink-based deposition by extrusion printing, spheroids were combined with a photo-crosslinkable methacrylamide-modified gelatin (gelMA) and Irgacure 2959. The influence of gelMA encapsulation, the printing process and photo-crosslinking conditions on spheroid viability, proliferation and vascularization were analyzed by live/dead staining, immunohistochemistry, gene expression analysis and sprouting analysis. Stable spheroid-laden constructs, allowing spheroid outgrowth, were achieved by applying 10 min UV-A photo-curing (365 nm, 4 mW cm(-2)), while the construct was incubated in an additional Irgacure 2959 immersion solution. Following implantation in ovo onto a chick chorioallantoic membrane, the prevascular engineered constructs showed anastomosis with the host vasculature. This study demonstrated (a) the potential of triculture prevascularized spheroids for application as multicellular building blocks, (b) the processability of the spheroid-laden gelMA bioink by extrusion bioprinting and (c) the importance of photo-crosslinking parameters post printing, as prolonged photo-curing intervals showed to be detrimental for the angiogenic potential and complete vascularization of the construct post printing.
Keywords
Biomedical Engineering, General Medicine, Biomaterials, Biochemistry, Bioengineering, Biotechnology, spheroids, vascular, endothelial cells, capillary, microvasculature, bioprinting, gelatin, MATRIX METALLOPROTEINASES, DIFFERENTIAL ADHESION, CELL, ANGIOGENESIS, GROWTH, MORPHOGENESIS, CONSTRUCTS, MIGRATION, CULTURE, MODEL, spheroids, vascular, endothelial cells, capillary, microvasculature, bioprinting, gelatin

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MLA
De Moor, Lise, et al. “Engineering Microvasculature by 3D Bioprinting of Prevascularized Spheroids in Photo-Crosslinkable Gelatin.” BIOFABRICATION, vol. 13, no. 4, 2021, doi:10.1088/1758-5090/ac24de.
APA
De Moor, L., Smet, J., Plovyt, M., Bekaert, B., Vercruysse, C., Asadian, M., … Declercq, H. (2021). Engineering microvasculature by 3D bioprinting of prevascularized spheroids in photo-crosslinkable gelatin. BIOFABRICATION, 13(4). https://doi.org/10.1088/1758-5090/ac24de
Chicago author-date
De Moor, Lise, Jasper Smet, Magaly Plovyt, Bieke Bekaert, Chris Vercruysse, Mahtab Asadian, Nathalie De Geyter, Sandra Van Vlierberghe, Peter Dubruel, and Heidi Declercq. 2021. “Engineering Microvasculature by 3D Bioprinting of Prevascularized Spheroids in Photo-Crosslinkable Gelatin.” BIOFABRICATION 13 (4). https://doi.org/10.1088/1758-5090/ac24de.
Chicago author-date (all authors)
De Moor, Lise, Jasper Smet, Magaly Plovyt, Bieke Bekaert, Chris Vercruysse, Mahtab Asadian, Nathalie De Geyter, Sandra Van Vlierberghe, Peter Dubruel, and Heidi Declercq. 2021. “Engineering Microvasculature by 3D Bioprinting of Prevascularized Spheroids in Photo-Crosslinkable Gelatin.” BIOFABRICATION 13 (4). doi:10.1088/1758-5090/ac24de.
Vancouver
1.
De Moor L, Smet J, Plovyt M, Bekaert B, Vercruysse C, Asadian M, et al. Engineering microvasculature by 3D bioprinting of prevascularized spheroids in photo-crosslinkable gelatin. BIOFABRICATION. 2021;13(4).
IEEE
[1]
L. De Moor et al., “Engineering microvasculature by 3D bioprinting of prevascularized spheroids in photo-crosslinkable gelatin,” BIOFABRICATION, vol. 13, no. 4, 2021.
@article{8721518,
  abstract     = {{To engineer tissues with clinically relevant dimensions by three-dimensional bioprinting, an extended vascular network with diameters ranging from the macro- to micro-scale needs to be integrated. Extrusion-based bioprinting is the most commonly applied bioprinting technique but due to the limited resolution of conventional bioprinters, the establishment of a microvascular network for the transfer of oxygen, nutrients and metabolic waste products remains challenging. To answer this need, this study assessed the potential and processability of spheroids, containing a capillary-like network, to be used as micron-sized prevascularized units for incorporation throughout the bioprinted construct. Prevascularized spheroids were generated by combining endothelial cells with fibroblasts and adipose tissue-derived mesenchymal stem cells as supporting cells. To serve as a viscous medium for the bioink-based deposition by extrusion printing, spheroids were combined with a photo-crosslinkable methacrylamide-modified gelatin (gelMA) and Irgacure 2959. The influence of gelMA encapsulation, the printing process and photo-crosslinking conditions on spheroid viability, proliferation and vascularization were analyzed by live/dead staining, immunohistochemistry, gene expression analysis and sprouting analysis. Stable spheroid-laden constructs, allowing spheroid outgrowth, were achieved by applying 10 min UV-A photo-curing (365 nm, 4 mW cm(-2)), while the construct was incubated in an additional Irgacure 2959 immersion solution. Following implantation in ovo onto a chick chorioallantoic membrane, the prevascular engineered constructs showed anastomosis with the host vasculature. This study demonstrated (a) the potential of triculture prevascularized spheroids for application as multicellular building blocks, (b) the processability of the spheroid-laden gelMA bioink by extrusion bioprinting and (c) the importance of photo-crosslinking parameters post printing, as prolonged photo-curing intervals showed to be detrimental for the angiogenic potential and complete vascularization of the construct post printing.}},
  articleno    = {{045021}},
  author       = {{De Moor, Lise and Smet, Jasper and Plovyt, Magaly and Bekaert, Bieke and Vercruysse, Chris and Asadian, Mahtab and De Geyter, Nathalie and Van Vlierberghe, Sandra and Dubruel, Peter and Declercq, Heidi}},
  issn         = {{1758-5082}},
  journal      = {{BIOFABRICATION}},
  keywords     = {{Biomedical Engineering,General Medicine,Biomaterials,Biochemistry,Bioengineering,Biotechnology,spheroids,vascular,endothelial cells,capillary,microvasculature,bioprinting,gelatin,MATRIX METALLOPROTEINASES,DIFFERENTIAL ADHESION,CELL,ANGIOGENESIS,GROWTH,MORPHOGENESIS,CONSTRUCTS,MIGRATION,CULTURE,MODEL,spheroids,vascular,endothelial cells,capillary,microvasculature,bioprinting,gelatin}},
  language     = {{eng}},
  number       = {{4}},
  pages        = {{20}},
  title        = {{Engineering microvasculature by 3D bioprinting of prevascularized spheroids in photo-crosslinkable gelatin}},
  url          = {{http://dx.doi.org/10.1088/1758-5090/ac24de}},
  volume       = {{13}},
  year         = {{2021}},
}

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