Marine sponges as natural scaffolds : decellularization by supercritical fluid technology and cellularization with osteoblasts for tissue engineering applications

摘要

Marine sponges possess remarkable structures, with highly interconnectedpores, making them excellent candidates as nature made scaffoldswith potential use in tissue engineering strategies. In additionthey also exhibit interesting high swelling degree and a skeleton composedmainly of collagen fibres, in some species with embedded siliceousor carbonate spicules. In the present study, several marinesponges (Dysidea avara, Chondrosia remiformis, Axinella damicornis, Petrosia ficiformis, Agelas oroides, Sarcotragus spinosulus, and Psammociniasp.) were decellularized by supercritical fluids, for further use asnatural scaffolds for osteoblasts. Supercritical fluid extraction was performedat 200 bar and 40 °C for 6 hours, in the presence of ethanol(40%), which was used as a co-solvent. The morphology of sponges,namely porosity interconnectivity and mean pore size, was analysed bySEM and micro-CT. To evaluate decellularization, sponges before andafter treatment were analysed in sections stained with Hematoxylin-Eosin. Cellularization experiments were performed in S. spinosulus,using Saos-2 cell line. Cells were seeded on the sponge, with furtherevaluation of cell morphology by SEM, cell viability by Live/dead cellstaining together with phalloidin.and cell proliferation by DNA quantification.The results of in vitro tests demonstrate the success of the proposedmethodology for the production of natural scaffolds from marinesponges, namely for Sarcotragus spinosulus.