3D Chitin Scaffolds of Marine Demosponge Origin for Biomimetic Mollusk Hemolymph-Associated Biomineralization <i>Ex-Vivo</i>
Marcin Wysokowski | Tomasz Machałowski | Iaroslav Petrenko | Christian Schimpf | David Rafaja | Roberta Galli | Jerzy Ziętek | Snežana Pantović | Alona Voronkina | Valentine Kovalchuk | Viatcheslav N. Ivanenko | Bert W. Hoeksema | Cristina Diaz | Yuliya Khrunyk | Allison L. Stelling | Marco Giovine | Teofil Jesionowski | Hermann Ehrlich
Structure-based tissue engineering requires large-scale 3D cell/tissue manufacture technologies, to produce biologically active scaffolds. Special attention is currently paid to naturally pre-designed scaffolds found in skeletons of marine sponges, which represent a renewable resource of biomaterials. Here, an innovative approach to the production of mineralized scaffolds of natural origin is proposed. For the first time, a method to obtain calcium carbonate deposition ex vivo, using living mollusks hemolymph and a marine-sponge-derived template, is specifically described. For this purpose, the marine sponge <i>Aplysin aarcheri</i> and the terrestrial snail <i>Cornu aspersum</i> were selected as appropriate 3D chitinous scaffold and as hemolymph donor, respectively. The formation of calcium-based phase on the surface of chitinous matrix after its immersion into hemolymph was confirmed by Alizarin Red staining. A direct role of mollusks hemocytes is proposed in the creation of fine-tuned microenvironment necessary for calcification ex vivo. The X-ray diffraction pattern of the sample showed a high CaCO<sub>3</sub> amorphous content. Raman spectroscopy evidenced also a crystalline component, with spectra corresponding to biogenic calcite. This study resulted in the development of a new biomimetic product based on ex vivo synthetized ACC and calcite tightly bound to the surface of 3D sponge chitin structure.
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