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SM ISO690:2012 MALCOVA, Tatiana, NACU, Viorel, ROJNOVEANU, Gheorghe, BIRGIT, Andree, HILFIKER, Andres. Successful decellularization of porcine aorta for acellular scaffold generation with the purpose of blood vessels tissue engineeing. In: Cercetarea în biomedicină și sănătate: calitate, excelență și performanță, Ed. 1, 20-22 octombrie 2021, Chişinău. Chișinău, Republica Moldova: 2021, p. 250. ISBN 978-9975-82-223-7 (PDF).. |
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Cercetarea în biomedicină și sănătate: calitate, excelență și performanță 2021 | ||
Conferința "Cercetarea în biomedicină și sănătate: calitate, excelență și performanță" 1, Chişinău, Moldova, 20-22 octombrie 2021 | ||
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Pag. 250-250 | ||
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Rezumat | ||
Background. Based on limited availability of autologous vascular grafts, the development of tissue-engineered blood vessels (TEBVs) has reached significant research interest in the last couples of years. Fabrication of TEBVs from decellulazed scaffolds seems to be an attractive research strategy. Objective of the study. To develop a method for the generation of cell-free vascular scaffolds by combining a detergent-based decellularization (DC) strategy with subsequent nuclease treatment. Material and Methods. Cryopreserved porcine aortae were treated with detergents for 48h under rotation (24h exposure to 0.5% SDS and 0.5% SDC, followed by 24h exposure to 1% TritonX-100) with/without following enzymatic digestion (48h exposure to 300 U/ml DNase I solution). The efficacy of DC was evaluated by 4’,6-Diamidino-2-phenylindole (DAPI) and hematoxylin and eosin (H&E) stainings. Results. H&E staining revealed no persisting cells in all groups, including the samples which were not treated with DNase. The DAPI stain of those specimens, however, uncovered substantial amounts of residual DNA. Additional enzymatic treatment with DNase led to such a reduction of residual DNA that DAPI positive structures could not be detected. Conclusion. A direct stain of DNA, e.g. DAPI, is much more sensitive measure for the presence of cell debris than the H&E stain. In addition, detergent-based decellularization technique per se is not sufficient for complete cell and debris removal and has to be combined with a DNase digestion. |
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Cuvinte-cheie blood vessel, tissue-engineering, decellularization, detergents, DNase, vas sanguin, inginerie tisulară, decelularizare, detergenti, DNaza |
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