Effect of three decellularisation protocols on the mechanical behaviour and structural properties of sheep aortic valve conduits |
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| Affiliation: | 1. Pediatric Urology Research Center, Section of Tissue Engineering and Stem Cells Therapy, Children''s Hospital Medical Center, Tehran University of Medical Sciences, Tehran, Iran;2. Cancer Research Center, Iranian Cancer Institute, Department of Pathology, Tehran University of Medical Sciences, Tehran, Iran;1. Department of Clinical Laboratory Diagnostics, Medical University of Bialystok, Bialystok, Poland;2. Department of Family Medicine and Community Nursing, Medical University of Bialystok, Bialystok, Poland;3. Department of Hematology, Clinical Hospital of the Medical University of Bialystok, Bialystok, Poland;1. I Department of Cardiology, Invasive Electrocardiology and Hypertension, Jagiellonian University Medical College, University, Krakow, Poland;2. Department of Invasive Cardiology, Electrotherapy and Angiology, Nowy Sacz Hospital, Nowy Sacz, Poland;1. Heart Institute, Faculty of Medicine, University of Pécs, Pécs, Hungary;2. Institute of Public Health and Health Promotion, Faculty of Health Sciences, University of Pécs, Pécs, Hungary;3. Department of Cardiac Surgery, Zala County Hospital, Hungary;4. Klinik für Kinder und Jugendliche, University of Erlangen-Nürnberg, Nürnberg, Germany;1. Department of Gastroenterology and Internal Medicine, Medical University of Bialystok, Bialystok, Poland;2. Department of Medical Pathomorphology, Medical University of Bialystok, Bialystok, Poland;3. Department of Gastroenterology and Internal Medicine, Regional Hospital, Bialystok, Poland |
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| Abstract: | PurposeTo determine the best method for decellularisation of aortic valve conduits (AVCs) that efficiently removes the cells while preserving the extracellular matrix (ECM) by examining the valvular and conduit sections separately.Material/methodsSheep AVCs were decellularised by using three different protocols: detergent-based (1% SDS + 1% SDC), detergent and enzyme-based (Triton + EDTA + RNase and DNase), and enzyme-based (Trypsin + RNase and DNase) methods. The efficacy of the decellularisation methods to completely remove the cells while preserving the ECM was evaluated by histological evaluation, scanning electron microscopy (SEM), hydroxyproline analysis, tensile test, and DAPI staining.ResultsThe detergent-based method completely removed the cells and left the ECM and collagen content in the valve and conduit sections relatively well preserved. The detergent and enzyme-based protocol did not completely remove the cells, but left the collagen content in both sections well preserved. ECM deterioration was observed in the aortic valves (AVs), but the ultrastructure of the conduits was well preserved, with no media distortion. The enzyme-based protocol removed the cells relatively well; however, mild structural distortion and poor collagen content was observed in the AVs. Incomplete cell removal (better than that observed with the detergent and enzyme-based protocol), poor collagen preservation, and mild structural distortion were observed in conduits treated with the enzyme-based method.ConclusionsThe results suggested that the detergent-based methods are the most effective protocols for cell removal and ECM preservation of AVCs. The AVCs treated with this detergent-based method may be excellent scaffolds for recellularisation. |
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| Keywords: | Aorta Heart valve Aortic conduit Acellular Sheep |
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