Production, Characterization and Biocompatibility of Marine Collagen Matrices from an Alternative and Sustainable Source: The Sea Urchin Paracentrotus lividus

Abstract

Collagen has become a key-molecule in cell culture studies and in the tissue engineering field. Industrially, the principal sources of collagen are calf skin and bones which, however, could be associated to risks of serious disease transmission. In fact, collagen derived from alternative and riskless sources is required, and marine organisms are among the safest and recently exploited ones. Sea urchins possess a circular area of soft tissue surrounding the mouth, the peristomial membrane (PM), mainly composed by mammalian-like collagen. The PM of the edible sea urchin Paracentrotus lividus therefore represents a potential unexploited collagen source, easily obtainable as a food industry waste product. Our results demonstrate that it is possible to extract native collagen fibrils from the PM and produce suitable substrates for in vitro system. The obtained matrices appear as a homogeneous fibrillar network (mean fibril diameter 30–400 nm and mesh < 2 μm) and display remarkable mechanical properties in term of stiffness (146 ± 48 MPa) and viscosity (60.98 ± 52.07 GPa·s). In vitro tests with horse pbMSC show a good biocompatibility in terms of overall cell growth. The obtained results indicate that the sea urchin P. lividus can be a valuable low-cost collagen source for mechanically resistant biomedical devices.

Caves Biodiversity in the Marine Area of Riviera d'Ulisse Regional Park, Italy: Grotta del Maresciallo Overview

“…the fish Apogon imberbis; the sponges Agelas oroides, Petrosia ficiformis, Chondrosia reniformis [9,12] and permits to identify species with great interest for their edibility as the fishes, Scorpaena notate, Phycis phycis, Conger conger, Sciaena umbra, Epinephelus marginatus, Epinephelus costae; the mollusca, Stramonita haemastoma, Arca noae; the crustacea Scyllarus arctus, Palaemon serratus and Parapandalus narval. Furthermore, we detected in the "Grotta del Maresciallo" cave a lot of species with various economic utility: against human breast cancer (MCF-7) and human neuroblastoma (SH-SY5Y) cell lines as for the spiny starfish, Marthasterias glacialis [44]; potential unexploited collagen source, easily obtainable as a food industry waste product, as for the edible sea urchin Paracentrotus lividus [45]; source of original natural substances belonging to two families of guanidine alkaloids, namely crambescins and crambescidins, which exhibit cytotoxic and antiviral activities, as for the orange-red encrusting sponge, Crambe crambe [46] and natural source of nanoparticles for the transdermal drug delivery of 17beta-estradiol-hemihydrate in hormone replacement therapy, as for the kidney-shaped sponge, Chondrosia reniformis [47]. Furthermore, in the "Grotta del Maresciallo" can be found species where were isolated actinobacterial strains and screened for antagonistic activity against various bacterial and fungal pathogens, as for the bath sponge, Spongia officinalis [48] and were isolated and purified acetylcholinesterase inhibitor isolated, as from the yellow encrusting anemone, Parazoanthus axinellae [49].…”

Deformation and fracture of echinoderm collagen networks

“…Sea urchin and starfish collagen were extracted from the PM and ABW, respectively, as previously described [24] with only slight modifications for the latter (see below). Briefly, both starfish ABW and sea urchin PM were dissected in small pieces, rinsed in artificial sea water (ASW), left in a hypotonic buffer (10 mM Tris, 0,1% EDTA) for 12 h (RT) and then in a decellularizing solution (10 mM Tris, 0,1% SDS) for 12 h (RT).…”

“…Membranes of the three different collagen types were prepared as previously described for sea urchin collagen matrices [24]. The collagen suspensions were centrifuged for 10 minutes at 50x g and then for 20 minutes at 2000x g. The pellet was re-suspended in autoclaved dH2O (final concentration 2mg/mL) and 8,7mL of this suspension were placed in 5cm x 3 cm rubber silicone molds and left dry overnight at +37 • C. The so obtained collagen membranes were then immersed in a EDC/NHS cross-linker solution (EDC 30 mM/NHS 15 mM in MES buffer 100 mM pH 5,5) for 4 h and then washed with PBS, dH2O and ethanol 70%.…”

“…In particular, collagen barrier-membranes of mammalian origin are typically used to facilitate proper tissue regrowth in anatomically separated compartments during guided tissue regeneration [25]. Marine collagen and MCTs in particular could provide an innovative and safer alternative to mammalian ones [26,24,27]. Moreover, other new intriguing applications could be derived from the use of collagen derived from echinoderms: for example to investigate cellular contractility as a biomechanics-related readout for phenotypic drug assessment [28] In this paper, we characterize the elastic tensile deformation and failure of cross-linked collagen networks derived from three echinoderms ( Fig.…”

Interfibrillar stiffening of echinoderm mutable collagenous tissue demonstrated at the nanoscale

“…A notable exception is the mutable collagenous tissue (MCT) of echinoderms (e.g., starfish, sea urchins, sea cucumbers), which undergoes rapid changes in stiffness under the control of the nervous system via ATP-independent mechanisms (10)(11)(12). MCT is ubiquitous in echinoderms (12), for example, in the dermis (skin) of sea cucumbers (13,14), in the compass depressor ligament (CDL) of sea urchins (15)(16)(17), and in the arms of feather stars (18). The presence of MCT enables functionally diverse behaviors; for example in starfish, MCT enables body wall stiffening during feeding on prey and it also enables irreversible body wall softening before arm autotomy as a defense against predation (12).…”