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Alternatives to Laboratory Animals - ATLA

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An In Vitro Perfusion System to Examine the Responses of Endothelial Cells to Simulated Flow and Inflammatory Stimulation

Natalia Y. Cockcroft, Oluwatobiloba Oke, Fiona Cunningham, Emma Bishop, Ian M. Fearon, Roman Zantl and Marianna D. Gaça

Atherosclerosis is a disease process which develops at the arterial branches and curvatures of medium to large arteries. Local haemodynamic flow patterns in these vessels play an essential role in the formation of atherosclerotic lesions. To simulate pro-atherogenic blood flow patterns, we have developed a perfusion system with the ability to simulate in vivo patterns of blood flow in vitro. In this system, human umbilical vein endothelial cells were seeded in y-shaped microslides, in which they were exposed to a variety of flow patterns. Besides being able to reproduce the disturbed flow involved in the development of pro-atherogenic events within the arterial wall, the system also permitted the assessment of the pre-conditioning/priming effect of oscillatory flow on endothelial cells. The system was further capable of integrating multi-endpoint assays relevant to cardiovascular disease. We show that oscillatory flow primed endothelial cells, making them more sensitive to subsequent treatments. The treatment of oscillatory flowprimed cells with TNFα resulted in the detection of enhanced levels of pro-inflammatory and chemoattractant factors such as IL-8 and MCP-1. These measurements were facilitated by the small volumes of medium circulating within the perfusion system. Oscillatory flow also altered the characteristics of monocyte adhesion to the endothelial layer. In summary, this system allows the monitoring of multiple endpoints and biomarkers, and provides an alternative to the use of in vivo and ex vivo models of cardiovascular disease.

Full text pdf 37(6), 657–669