Noninvasive Analysis of Synthetic and Decellularized Scaffolds for Heart Valve Tissue Engineering

Microcomputed tomography (mu-CT) is a nondestructive,
high-resolution, three-dimensional method of analyzing objects. The
aim of this study was to evaluate the feasibility of using mu-CT as
a noninvasive method of evaluation for tissue-engineering
applications. The polyurethane aortic heart valve scaffold was
produced using a spraying technique. Cryopreserved/thawed homograft
and biological heart valve were decellularized using a detergent
mixture. Human endothelial cells and fibroblasts were derived from
saphenous vein segments and were verified by immunocytochemistry.
Heart valves were initially seeded with fibroblasts followed by
colonization with endothelial cells. Scaffolds were scanned by a
mu-CT scanner before and after decellularization as well as after
cell seeding. Successful colonization was additionally determined
by scanning electron microscopy (SEM) and immunohistochemistry
(IHC). Microcomputed tomography accurately visualized the complex
geometry of heart valves. Moreover, an increase in the total volume
and wall thickness as well as a decrease in total surface was
demonstrated after seeding. A confluent cell distribution on the
heart valves after seeding was confirmed by SEM and IHC. We
conclude that mu-CT is a new promising noninvasive method for
qualitative and quantitative analysis of tissue-engineering
processes. ASAIO Journal 2013;59:169-177.