Abstract
Aortic dysfunctions (aneurysm, aortitis) lead to the most serious conditions related to aortic wall with life-threatening complications. The most common modality of management for such conditions is replacement (diseased part) of aorta by a larger diameter stent (reconstructive vascular surgery) which in itself is a big trial. The most natural way is to use a re-endothelized scaffold. Developing a scaffold with biomimetic properties is an experimental aim for most of the scientists and surgeons. We aim to structure a strategy to overcome the well-known problems associated with aorta. In this study, we plan to remold a larger diameter blood vessel such as aorta from xenogeneic origin using different protocols to decellularize and comparing them with normal aorta. The chemicals and enzymes used for bovine aorta decellularization are 1% SDS (group II), 70% ethanol + 0.25% trypsin (group III), 70% ethanol (group IV), and 0.25% trypsin (group V). Group I served as control (without decellularization). Histology and SEM study were conducted for cellular presence/absence in all scaffolds. Later, the scaffolds were coated with the fibrin glue (FG) and endothelial cells were proliferated over them. 3D images were taken showing the remolding of the endothelial cells on FG-coated surfaces. The re-endothelization was confirmed by lectin and vWF+/+ expression. Graft elasticity and burst pressure were confirmed by biomechanical tensile testing. Further, the absence of host tissue DNA and presence of cellular DNA after re-endothelialization were confirmed by PicoGreen assay. The acceptability for metabolically active cellular proliferation on scaffolds and its non-toxicity were proved by cell viability assay. Current findings accomplish that larger diameter aorta extracellular matrix scaffold (group II) can be fabricated and re-endothelialized to develop non-thrombotic surfaces with improved graft patency with promising results compared to other fabricated scaffold groups.
Highlights
The aorta is the largest blood vessel in the body
The samples in groups I, II, IIA and IIB (n = 5) were placed in lysis buffer for 15 min and macerated. 30 μL of each sample was obtained in 4 mL cuvette and 2.96 mL of distilled water was added followed by DNA samples incubation for 10 min
The absorbance was measured at 260 nm with dsDNA levels quantified by a PicoGreen fluorescent assay to confirm the occurence of residual tissue ds-DNA in decellularized group
Summary
The aorta is the largest blood vessel in the body. It arises from the left ventricle of the heart and is divided into four parts: the ascending aorta, aortic arch, thoracic and abdominal part of descending aorta. It is almost a few centimeters (cm) long and has a circumferential diameter of approximately 2.54 cm. The intima, the innermost layer (basal lamina) provides a smooth surface for blood to flow across. The media, the middle layer with smooth muscles comprising elastic fibers that allows the aorta to expand and contract
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