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Alobaidan, Alsobaie, Yang, and Musa: Investigation of perfused platelet activation using a 3D tissue-engineered medial layer

Introduction: Haemostasis is a highly regulated physiological process of which platelets play a critical role in forming a blood clot, to prevent haemorrhage. The prothrombotic properties of the vasculature facilitate platelet activation upon damage whereas its antithrombotic properties prevent platelet activation, adhesion and aggregation.

Objectives: To fabricate a 3-dimensional (3D) medial layer by seeding rat dermal fibroblast cells (RDFCs) in collagen hydrogel to mimic smooth muscle cells in collagen hydrogel and a blood flow chamber to enable investigation of the effect of culture time period and shear stress on platelet activation.

Materials and methods: RDFCs were cultured within collagen type hydrogel, to mimic a component of the blood vessel construct, the tunica media. A blood flow chamber was constructed using polydimethylsiloxane (PDMS) mould to fit with the construct (dimensions 7.3 × 3 × 1.5 cm), forming a rhombic-shaped gasket. This chamber provided physiologically based flow regimes and enabled examination of platelet activation under flow conditions. Washed human platelets were perfused into the closed perfusion system containing the medial layer cultured for 2 and 7 days.

Results: RDFCs seeded in collagen hydrogel favourably survived within the 3D layer construct. Platelet aggregation is culture time dependent since perfused platelets aggregate quicker upon exposure to 7 days cultured RDFCs in collagen than 2 days culture.

Conclusions: Medial construct containing RDFCs were shown to activate platelets highlighting the release of neocollagen. This in vitro perfusion chamber is a potential tool for haemostasis and platelet activation studies.

Acknowledgements: Ying Yang and Faiza Musa for supervising and support. Dr Waleed Alshaqha for supervising the project trip.




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