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In-Vitro Functionality Assessment of a New Tissue-Engineered Valve Substitute
Catherine Tremblay1, Jean Ruel1, Véronique Laterreur1, Karine Vallières2, Jean-Michel Bourget2, Maxime Y. Tondreau2, Dan Lacroix2, Lucie Germain2, François A. Auger2.
1Université Laval, Quebec City, QC, Canada, 2Centre LOEX, Quebec City, QC, Canada.

BACKGROUND: Aortic valve replacement surgeries are usually performed using mechanical prostheses or tissue valves. While those prostheses have greatly evolved in the past decades, many serious drawbacks, such as lifelong anticoagulant therapies or accelerated calcification of the substitutes, often lower the life quality of the patients. Moreover, they still remain foreign materials to the body and are not able to remodel with their environment. The search for an entirely biological substitute that would eliminate those drawbacks is still relevant. Therefore, the purpose of this study is to develop a construction technique using the self-assembly method for a new biological heart valve substitute and to assess its functionality in a bioreactor.
METHODS: Dermal fibroblasts were cultured for 26 days in medium supplemented with ascorbic acid in order to create thin manipulable sheets. Cell sheets were stacked to create a thick plane tissue. Custom-made sets of templates were designed to help with the construction of the valve in its three-dimensional configuration. The valve was then placed in a custom built bioreactor for three days to assess its functionality under a large variety of pulsed flow.
RESULTS: The shape of the 23 mm diameter tissue-engineered valve resembled that of a native aortic valve. The three leaflets presented a symmetrical configuration and adequately touched each other to create a tight and continuous contact surface. The valve was progressively exposed to pulsed flow with frequency ranging from 0.33 Hz to 1 Hz. Flowrates were also modulated from very mild conditions to physiological-like flow rates. The valve maintained its shape throughout the whole experiment. Histological analysis of the valve at all steps of the construction technique presented a dense extracellular matrix and well-distributed cells.
CONCLUSIONS: The self-assembly method seems a promising approach for the construction of a new heart valve substitute. The use of different sets of templates helped to establish a repeatable construction technique able to create a complex three-dimensional structure with only a plane tissue to begin with. In short-term tests, the new heart valve substitute was able to sustain a large variety of flow and pressure waves while maintaining its structural integrity.

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