Abstract
Angiogenesis plays a pivotal role in development and tissue growth, as well as in pathological conditions such as cancer. Being able to understand the basic mechanisms involved in the vascularization of tissues and angiogenic network formation provides a window to advance the development of in vitro tissue models and enhance tissue engineering applications. In this study, we leveraged a novel microfluidic-based three dimensional (3D) bioprinting technology and alginate-collagen type I (AGC) bioink, to develop a 3D bioprinting strategy to enable the biofabrication of complex angiogenic networks within the 3D structure. These networks were comprised of simian vacuolating virus 40 (SV40) transformed adult rat brain endothelial cell (SV-ARBEC)-laden hydrogel rings. With mechanical properties relevant for vascular tissue engineering applications, these bioprinted constructs formed spontaneous vascular networks, reminiscent of anisotropic tissue-like structures, while retaining high cellular viability. The vascular network formation was accompanied by extracellular matrix (ECM) remodeling, confirming sequential SV-ARBEC mediated collagen type I fiber deposition and reorganization. Treatment with broad spectrum matrix metalloproteinase (MMP) inhibitor supressed SV-ARBEC angiogenic sprouting, highlighting requirements of ECM remodeling in angiogenic network formation. This novel 3D microfluidic bioprinting technology and biocompatible AGC hydrogel fiber rings supported robust SV-ARBEC angiogenesis and corresponding ECM remodeling, allowing us to present a strategy suitable to advancing applications in vascular research and supporting the further development of disease models, novel testing beds for drug discovery and tissue engineering applications.
| Original language | English |
|---|---|
| Article number | e00258 |
| Journal | Bioprinting |
| Volume | 30 |
| DOIs | |
| State | Published - Apr 2023 |
Funding
We would like to thank Claudie Charlebois and Klaudia Baumann for maintaining SV-ARBEC cultures for the bioprinting studies. We would like to thank Melissa Hewitt for providing support in fiber processing and staining. We would like to thank Dr. Robert Monette, Dr. Ana Magalhaes and Dongling Zhang for providing technical support on confocal imaging. We would also like to thank Danielle Abdul Nour for her help with preparing hydrogel samples for mechanical testing. Funding was provided by the NRC's Ideation Programs – New Beginnings Fund. The Aspect Biosystems RX1 Bioprinter was acquired by the NRC through the Built in Canada Innovation Program. We would like to thank the Aspect Biosystems team for providing technical and experimental support. The Biomomentum Mach-1 mechanical tester used in the study was acquired with funds from the Canadian Foundation for Innovation [JPS: CFI-37732; 2020–2022 ] with matching funds from the Ministry of Economic Development, Job Creation and Trade Ontario Research Fund . JN was supported by a Canada Graduate Scholarships – Master's program from the Natural Sciences and Engineering Research Council of Canada (NSERC) , and an Ontario Graduate Scholarship . L.B.M.-G. was supported by the Natural Sciences and Engineering Research Council (Discovery Grant of Canada ( RGPIN-2021-04185 ). JPS was supported by the Natural Sciences and Engineering Research Council Discovery Grant of Canada ( RGPIN-2017-04593 ). The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Jean-Philippe St-Pierre reports financial support and equipment, drugs, or supplies were provided by Canadian Foundation for Innovations. Jean-Philippe St-Pierre reports financial support and equipment, drugs, or supplies were provided by Ontario Ministry of Economic Development Job Creation and Trade. Leila Mostaco-Guidolin reports financial support was provided by Natural Sciences and Engineering Research Council of Canada. Jean-Philippe St-Pierre reports financial support was provided by Natural Sciences and Engineering Research Council of Canada.We would like to thank Claudie Charlebois and Klaudia Baumann for maintaining SV-ARBEC cultures for the bioprinting studies. We would like to thank Melissa Hewitt for providing support in fiber processing and staining. We would like to thank Dr. Robert Monette, Dr. Ana Magalhaes and Dongling Zhang for providing technical support on confocal imaging. We would also like to thank Danielle Abdul Nour for her help with preparing hydrogel samples for mechanical testing. Funding was provided by the NRC's Ideation Programs – New Beginnings Fund. The Aspect Biosystems RX1 Bioprinter was acquired by the NRC through the Built in Canada Innovation Program. We would like to thank the Aspect Biosystems team for providing technical and experimental support. The Biomomentum Mach-1 mechanical tester used in the study was acquired with funds from the Canadian Foundation for Innovation [JPS: CFI-37732; 2020–2022] with matching funds from the Ministry of Economic Development, Job Creation and Trade Ontario Research Fund. JN was supported by a Canada Graduate Scholarships – Master's program from the Natural Sciences and Engineering Research Council of Canada (NSERC), and an Ontario Graduate Scholarship. L.B.M.-G. was supported by the Natural Sciences and Engineering Research Council (Discovery Grant of Canada (RGPIN-2021-04185). JPS was supported by the Natural Sciences and Engineering Research Council Discovery Grant of Canada (RGPIN-2017-04593).
Keywords
- 3D bioprinting
- Alginate
- Angiogenesis
- Bioink
- Collagen
- ECM Remodeling
- Microfluidics
- Vascular networks