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

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.

Naturally Formed Chitinous Skeleton Isolated from the Marine Demosponge Aplysina fistularis as a 3D Scaffold for Tissue Engineering

“…Marine sponges are a remarkable donors of renewable biological materials [ 28 , 31 ]. Especially Verongida (Porifera) in particular was established as a good potential source of bioactive compounds (e.g., aeroplysinin-1 or isofistularin-3) [ 4 , 32 , 33 ] and chitinous skeletons [ 34 , 35 , 36 , 37 , 38 ].…”

Electrochemical Approach for Isolation of Chitin from the Skeleton of the Black Coral Cirrhipathes sp. (Antipatharia)

“…Thus, we obtained a membranous organic matrix, with regular pores up to 100 µm large. Calcofluor white staining of this matter ( Figure 7B) allows us to assume, with a high probability, the chitinous nature of the matrix, taking into account previously published results on chitin identification using this broadly applied technique [3][4][5]9,10,13,16,[46][47][48][49][50][51][52][53][54][55][56][57][58]. For indisputable identification of chitin, we used infrared spectroscopy (ATR FT-IR), the chitinase digestion test, and ESI-MS-based analytics as represented below.…”

“…Consequently, its extraction from such biocomposites is fraught with a number of methodological difficulties that must be overcome using different approaches. Nowadays, chitin of marine invertebrate origin is commonly isolated via two types of extraction process: chemical or biological [15,16]. In brief, the chemical treatment requires three main steps, namely deproteinization, demineralization and depigmentation.…”

“…Thus, electrolysis-assisted isolation of chitin has not received much attention over traditionally used extraction procedures. As we recently showed with experiments on chitin from the skeleton of the marine sponge Aplysina aerophoba (Nardo, 1833), the electrolysis process can be very flexible in terms of electrochemical conditions [16]. Thus, there is possible scope for the improvement of this overlooked approach-for example, by the application of different electrolytes.…”

Functionalization of 3D Chitinous Skeletal Scaffolds of Sponge Origin Using Silver Nanoparticles and Their Antibacterial Properties

“…Today, bromotyrosines are recognized as multi-targeted marine drugs with broad fields of applications (i.e., as antitumorigenic and antimetastatic agents [55,56]). In order to isolate pure chitinous scaffolds from verongiid demosponges, diverse methodological approaches based on chemical, electrochemical and enzymatic treatments were reported (see for overview [36,57]). Nonetheless, the alkali treatment of verongiid skeletons is most commonly used [20,36,40] and can be regulated with respect to residual bromine concentration in the chitinous matrices.…”