The aortic biosheet valve extracted specimens were fixed with 10 % formalin, embedded in paraffin, sliced into longitudinal sections, and finally stained with hematoxylin–eosin (H&E), Elastica-van Giessen, Masson’s trichrome (collagen stain), and von Kossa (calcium stain). In addition, a few sections of the biosheet were also stained for α-smooth muscle actin (α-SMA) using immunohistochemical techniques; the protein was detected using monoclonal antibodies (Abcam, Cambridge, UK). The histological analyses were not blinded. Only one pathologist performed the pathological evaluation.

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