Electrochemical method for isolation of chitinous 3D scaffolds from cultivated Aplysina aerophoba marine demosponge and its biomimetic application

Krzysztof Nowacki,Izabela Stępniak,Jerzy Ziętek,Tomasz Machałowski,Snežana Pantović,Andreas Richter,Teofil Jesionowski,Alona Voronkina,Michael Gelinsky,Valentine Kovalchuk,Roberta Galli,Yvonne Joseph,Christoph Schimpf,David Rafaja,Yuliya Khrunyk,Viatcheslav N Ivanenko ,Marcin Wysokowski,Enrico Langer,Iaroslav Petrenko,Hermann Ehrlich

doi:10.1007/s00339-020-03533-2

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.

Highlights

In recent decades, the synthesis and application of 3D biopolymer-based scaffolds represent one of the new trends in environmental science and technology
The photographs of the sample taken right after pretreatment step demonstrated a well-preserved original bio-architecture of A. aerophoba sponge
This image illustrates dark-brown, semitransparent and cellfree chitinous scaffold, the spatial structure of which can be characterized as typical branched network of chitinous tubes of the sponge origin

Summary

Introduction

The synthesis and application of 3D biopolymer-based scaffolds represent one of the new trends in environmental science and technology. Chitin is one of the most abundant polysaccharides of natural origin, obtained mainly from crustaceans, it can be found in representatives of diatoms, sponges, mollusks, tubeworms, insects and arachnids [17,18,19,20,21,22] This biopolymer is composed of β-(1,4)-N-acetyl-dglucosamine units and plays a crucial role in the formation of both soft and mineralized skeletal structures in invertebrates requiring rigidity and mechanical strength [23, 24].

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Conclusion