Abstract
Quantitative assessment of functional perfusion capacity and vessel architecture is critical when validating biomaterials for regenerative medicine purposes and requires high-tech analytical methods. Here, combining two clinically relevant imaging techniques, (magnetic resonance imaging; MRI and microcomputed tomography; MicroCT) and using the chorioallantoic membrane (CAM) assay, we present and validate a novel functional and morphological three-dimensional (3D) analysis strategy to study neovascularization in biomaterials relevant for bone regeneration. Using our new pump-assisted approach, the two scaffolds, Optimaix (laminar structure mimicking entities of the diaphysis) and DegraPol (highly porous resembling spongy bone), were shown to directly affect the architecture of the ingrowing neovasculature. Perfusion capacity (MRI) and total vessel volume (MicroCT) strongly correlated for both biomaterials, suggesting that our approach allows for a comprehensive evaluation of the vascularization pattern and efficiency of biomaterials. Being compliant with the 3R-principles (replacement, reduction and refinement), the well-established and easy-to-handle CAM model offers many advantages such as low costs, immune-incompetence and short experimental times with high-grade read-outs when compared to conventional animal models. Therefore, combined with our imaging-guided approach it represents a powerful tool to study angiogenesis in biomaterials.
Highlights
Tissue engineering often utilizes suitable biomaterials intended to guide and stimulate the healing process of the body[1,2,3,4]
The use of the chorioallantoic membrane (CAM) as an in vivo bioreactor allows for testing the biocompatibility of biomaterials[28], assessing bone repair in injured bone cylinders from human femoral heads placed on the CAM by MicroCT29, or studying in vivo functional perfusion capacity of tissue-engineered constructs (TECs) by MRI9,14 while keeping cells and organs alive
This was evident in the vascular architecture, which had developed from a simple structure with a few branches to a more complex network of vessels
Summary
Tissue engineering often utilizes suitable biomaterials intended to guide and stimulate the healing process of the body[1,2,3,4]. Most of the studies using MRI and MicroCT for vascular imaging and quantification were performed in large (e.g. sheep25) and small (e.g. rabbit[27] and rats24) animal models. Their application is linked to high costs, animal burden and often very time-consuming experiments. The use of the CAM as an in vivo bioreactor allows for testing the biocompatibility of biomaterials[28], assessing bone repair in injured bone cylinders from human femoral heads placed on the CAM by MicroCT29, or studying in vivo functional perfusion capacity of TECs by MRI9,14 while keeping cells and organs alive
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