Ultrasound-Based Three-Dimensional Microangiography for Repeated Noninvasive Imaging of Neovascularization

Fredrik Karl Erik Guldbæk-Svensson, Mathias Thygesen, Teresa Haugaard Nielsen, Trine Werenberg Mikkelsen, Ming Ding, Michael Pedersen, Henrik Lauridsen*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

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Neovascularization plays an essential part in the progression of several diseases as well as recovery processes such as angiogenesis guided neurogenesis after a spinal cord injury. To visualize and quantify neovascularization in an experimental setup it is desirable to apply a non-invasive, repeatable and harmless technique. Here we apply a newly developed ultrasound based technique to construct angiographies and quantify neovascularization in the regenerating spinal cord of a regeneration competent animal model, the Mexican axolotl. We measured vessel volume fraction prior to the induction of contusion and transection spinal cord injury and repeated this measurement directly after injury and at 3, 6, 12 and 63 days after injury. Whereas neither of the injury types resulted in statistically significant differences in vessel volume fraction relative to sham operated animals there was a statistically significant increase in neovascularization over time in all groups. Additionally, vessel volume fraction at the final time point (63 days after injury) was quantified with micro-CT imaging after vascular perfusion with a contrast agent, confirming no statistically significant difference in neovascularization between injury types. Ex vivo vessel volume fraction measured by micro-CT was significantly different from the in vivo ultrasound based measurement at the same time point. This is likely a result of incomplete vascular perfusion with the contrast agent before micro-CT imaging, which was supported by subsequent histological evaluation. In summary, the results suggest that the ultrasound based angiographic procedure we demonstrate here is applicable to visualize and quantify neovascularization in a non-invasive and harmless fashion in longitudinal experiments circumventing the limitations of contrast agent dependent techniques.
Original languageEnglish
Article number3396080
JournalContrast Media & Molecular Imaging
Publication statusPublished - Jan 2023


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