Electrochemical method for isolation of chitinous 3D scaffolds from cultivated Aplysina aerophoba marine demosponge and its biomimetic application
Krzysztof Nowacki, Izabela Stępniak, Tomasz Machałowski, Marcin Wysokowski, Iaroslav Petrenko, Christoph Schimpf, David Rafaja, Enrico Langer, Andreas Richter, J. Ziêtek, Snežana Pantović, Alona Voronkina, Valentine Kovalchuk, Viatcheslav N. Ivanenko, Yuliya Khrunyk, Roberta Galli, Yvonne Joseph, Michael Gelinsky, Teofil Jesionowski, Hermann Ehrlich
Abstract
Abstract Three-dimensional (3D) biopolymer-based scaffolds including chitinous matrices have been widely used for tissue engineering, regenerative medicine and other modern interdisciplinary fields including extreme biomimetics. In this study, we introduce a novel, electrochemically assisted method for 3D chitin scaffolds isolation from the cultivated marine demosponge Aplysina aerophoba which consists of three main steps: (1) decellularization, (2) decalcification and (3) main deproteinization along with desilicification and depigmentation. For the first time, the obtained electrochemically isolated 3D chitinous scaffolds have been further biomineralized ex vivo using hemolymph of Cornu aspersum edible snail aimed to generate calcium carbonates-based layered biomimetic scaffolds. The analysis of prior to, during and post-electrochemical isolation samples as well as samples treated with molluscan hemolymph was conducted employing analytical techniques such as SEM, XRD, ATR–FTIR and Raman spectroscopy. Finally, the use of described method for chitin isolation combined with biomineralization ex vivo resulted in the formation of crystalline (calcite) calcium carbonate-based deposits on the surface of chitinous scaffolds, which could serve as promising biomaterials for the wide range of biomedical, environmental and biomimetic applications.