Development of a human 3D tissue-engineered blood vessel model for the study of haemostasis

Musa, Faiza

Abstract

In this project, we have designed, constructed, and validated a human tissue-engineered blood vessel construct (TEBV) to assess whether it could be used as a new model system in which to study thrombus formation under physiologically relevant conditions.
A layer-by-layer fabrication technique was adopted to fabricate the TEBV. This allowed for the biological properties of both the medial and intimal layers of the construct to be assessed individually, as well as in combination as the full TEBV. The TEBV model was shown to mimic the anatomical structure and cellular phenotype of the native human artery. In addition, a novel technique for quantitatively assessing the pro- and anti-aggregatory properties of these constructs was developed, utilising real-time measurements of cytosolic Ca2+ signalling to assess real-time human platelet activation when exposed to the tissue-engineered blood vessels constructs. The real-time measurement of cytosolic Ca2+ signalling of human platelets when exposed to the TEBV models was shown to be a sensitive technique to assess the haemostatic properties of the 3-dimensional (3D) TEBV to validate the physiological relevance of the construct. Experiments conducted with this novel methodology, alongside other traditional platelet function assays, demonstrated that our TEBV had an anti-aggregatory intimal layer and a pro-aggregatory medial layer, consistent with the haemostatic functions of this blood vessel layers in vivo.
We have also established an ex vivo ferric chloride (FeCl3) arterial injury model to be used as an alternative to intravital microscopy study of in vivo thrombus formation in mice models. Treatment of the TEBV with FeCl3 elicited a significant increase in platelet aggregation upon the surface of the construct when these cells were perfused over the construct at arterial shear stresses. By using this perfusion system, experiments also provided initial evidence that the use of the general anaesthetic ketamine, in intravital microscopy experiments may interfere with thrombus formation, and therefore could affect the validity of results seen in previous in vivo studies. In conclusion, we have
successfully created a TEBV that is able to replicate the functional properties of the native vessel and which may be useful as a novel model to study human thrombus formation.

Citation

Musa, F. (2018). Development of a human 3D tissue-engineered blood vessel model for the study of haemostasis. (Thesis). Keele University. Retrieved from https://keele-repository.worktribe.com/output/411162

• Thesis Type: Thesis
• Publicly Available Date: May 26, 2023
• Public URL: https://keele-repository.worktribe.com/output/411162
• Award Date: 2018-06