Abstract
BackgroundTissue engineering has emerged as a promising alternative for small-diameter vascular grafts. The aim of this study was to determine the feasibility of using decellularized aortae of fetal pigs (DAFPs) to construct tissue-engineered, small-diameter vascular grafts and to test the performance and application of DAFPs as vascular tissue-engineered scaffolds in the canine arterial system.MethodsDAFPs were prepared by continuous enzymatic digestion. Canine vascular endothelial cells (ECs) were seeded onto DAFPs in vitro and then the vascular grafts were cultured in a custom-designed vascular bioreactor system for 7 days of dynamic culture following 3 days of static culture. The grafts were then transplanted into the common carotid artery of the same seven dogs from which ECs had been derived (two grafts were prepared for each dog with one as a backup; therefore, a total of 14 tissue-engineered blood vessels were prepared). At 1, 3, and 6 months post-transplantation, ultrasonography and contrast-enhanced computed tomography (CT) were used to check the patency of the grafts. Additionally, vascular grafts were sampled for histological and electron microscopic examination.ResultsTissue-engineered, small-diameter vascular grafts can be successfully constructed using DAFPs and canine vascular ECs. Ultrasonographic and CT test results confirmed that implanted vascular grafts displayed good patency with no obvious thrombi. Six months after implantation, the grafts had been remodeled and exhibited a similar structure to normal arteries. Immunohistochemical staining showed that cells had evenly infiltrated the tunica media and were identified as muscular fibroblasts. Scanning electron microscopy showed that the graft possessed a complete cell layer, and the internal cells of the graft were confirmed to be ECs by transmission electron microscopy.ConclusionsTissue-engineered, small-diameter vascular grafts constructed using DAFPs and canine vascular ECs can be successfully transplanted to replace the canine common carotid artery. This investigation potentially paves the way for solving a problem of considerable clinical need, i.e., the requirement for small-diameter vascular grafts.
Highlights
Tissue engineering has emerged as a promising alternative for small-diameter vascular grafts
Identification of canine vascular endothelial cells (ECs) and morphology of Decellularized aorta of fetal pigs (DAFP) Immunohistochemistry and immunofluorescence showed positive von Willebrand factor (vWF) staining in most cells (Fig. 2)
Histological staining of DAFPs showed that the extracellular matrix remained intact; no nuclei or intact cells were present (Fig. 3A-b)
Summary
Tissue engineering has emerged as a promising alternative for small-diameter vascular grafts. The aim of this study was to determine the feasibility of using decellularized aortae of fetal pigs (DAFPs) to construct tissueengineered, small-diameter vascular grafts and to test the performance and application of DAFPs as vascular tissueengineered scaffolds in the canine arterial system. Tissue engineering has emerged as a promising alternative for producing small-diameter vascular grafts [4]. Compared with synthetic polymer-based scaffolds, natural polymers present a biologically active environment to cells and promote excellent cell adhesion and growth [7]. Decellularized tissue-engineered vascular grafts have been widely used as natural scaffolds to produce arterial conduits that provide ideal biomechanical properties and cell compatibility [10, 11]. Böer et al showed that intensified decellularization of equine carotid arteries generated highly suitable matrix scaffolds for vascular tissue engineering [12]. The in vivo application of tissue-engineered vascular grafts has not been widely investigated
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