Isolation, Characterization and Biological Evaluation of Jellyfish Collagen for Use in Biomedical Applications

Fibrillar collagens are the more abundant extracellular proteins. They form a metazoan-specific family, and are highly conserved from sponge to human. Their structural and physiological properties have been successfully used in the food, cosmetic, and pharmaceutical industries. On the other hand, the increase of jellyfish has led us to consider this marine animal as a natural product for food and medicine. Here, we have tested different Mediterranean jellyfish species in order to investigate the economic potential of their collagens. We have studied different methods of collagen purification (tissues and experimental procedures). The best collagen yield was obtained using Rhizostoma pulmo oral arms and the pepsin extraction method (2–10 mg collagen/g of wet tissue). Although a significant yield was obtained with Cotylorhiza tuberculata (0.45 mg/g), R. pulmo was used for further experiments, this jellyfish being considered as harmless to humans and being an abundant source of material. Then, we compared the biological properties of R. pulmo collagen with mammalian fibrillar collagens in cell cytotoxicity assays and cell adhesion. There was no statistical difference in cytotoxicity (p > 0.05) between R. pulmo collagen and rat type I collagen. However, since heparin inhibits cell adhesion to jellyfish-native collagen by 55%, the main difference is that heparan sulfate proteoglycans could be preferentially involved in fibroblast and osteoblast adhesion to jellyfish collagens. Our data confirm the broad harmlessness of jellyfish collagens, and their biological effect on human cells that are similar to that of mammalian type I collagen. Given the bioavailability of jellyfish collagen and its biological properties, this marine material is thus a good candidate for replacing bovine or human collagens in selected biomedical applications

Isolation, Characterization and Biological Evaluation of Jellyfish Collagen for Use in Biomedical Applications

Fibrillar collagens are the more abundant extracellular proteins. They form a metazoan-specific family, and are highly conserved from sponge to human. Their structural and physiological properties have been successfully used in the food, cosmetic, and pharmaceutical industries. On the other hand, the increase of jellyfish has led us to consider this marine animal as a natural product for food and medicine. Here, we have tested different Mediterranean jellyfish species in order to investigate the economic potential of their collagens. We have studied different methods of collagen purification (tissues and experimental procedures). The best collagen yield was obtained using Rhizostoma pulmo oral arms and the pepsin extraction method (2–10 mg collagen/g of wet tissue). Although a significant yield was obtained with Cotylorhiza tuberculata (0.45 mg/g), R. pulmo was used for further experiments, this jellyfish being considered as harmless to humans and being an abundant source of material. Then, we compared the biological properties of R. pulmo collagen with mammalian fibrillar collagens in cell cytotoxicity assays and cell adhesion. There was no statistical difference in cytotoxicity (p > 0.05) between R. pulmo collagen and rat type I collagen. However, since heparin inhibits cell adhesion to jellyfish-native collagen by 55%, the main difference is that heparan sulfate proteoglycans could be preferentially involved in fibroblast and osteoblast adhesion to jellyfish collagens. Our data confirm the broad harmlessness of jellyfish collagens, and their biological effect on human cells that are similar to that of mammalian type I collagen. Given the bioavailability of jellyfish collagen and its biological properties, this marine material is thus a good candidate for replacing bovine or human collagens in selected biomedical applications

Biochemical and biological characterizations of jellyfish collagens

Ce travail a été entrepris dans le but de caractériser biochimiquement et biologiquement les collagènes de méduse. l’objectif de notre travail a été d’évaluer ce matériel en vue de sa valorisation pour l’élaboration de biomatériaux à usage médical. dans un premier temps, nous avons optimisé les techniques d’extractions des collagènes de quatre espèces de méduses : rhizostoma pulmo, cotylorhiza tuberculata, pelagia noctiluca et aurelia aurita. l’étude de la stabilité thermique par dichroïsme circulaire des collagènes nous a montré que la température de dénaturation des collagènes de rhizostoma pulmo était de 28,9°c. nous avons choisi de réaliser la réticulation des collagènes de méduse par la méthode des carbodiimides (edc/nhs). cette méthode nous a permis d’augmenter la température de dénaturation jusqu’à 33°c (versus 28,9°c). l’étude de l’interaction entre les cellules d’origine humaine (mg63 et fibroblastes) et les collagènes de méduses, nous a permis de démontrer qu’elles étaient capables d’adhérer aux collagènes de méduses natifs ou dénaturés. des immunomarquages de la vinculine des cellules mg63 et des fibroblastes sur collagènes de méduse natifs ou dénaturés nous ont permis de caractériser les adhésions mises en places par ces deux types cellulaires. l’analyse des milieux de culture des cellules mg63 et des fibroblastes par zymographie sur collagènes de méduse natifs ou dénaturés nous a permis de mettre en évidence que des mmps d’origine humaine, seraient capables de dégrader les collagènes de méduses. nous pouvons dans ce cas estimer que le collagène de méduse serait un bon substrat pour l’élaboration de biomatériaux résorbablesThis work was undertaken to characterize biochemically and biologically jellyfish collagen. the aim of our study was to evaluate the material for the development of biomaterials for medical use. initially, we optimized the technical extraction of collagen from four species of jellyfishes: rhizostoma pulmo, cotylorhiza tuberculata, pelagia noctiluca and aurelia aurita. the study of thermal stability by circular dichroism of collagen has shown that the denaturation temperature of rhizostoma pulmo collagens was 28.9 ° c. we chose to achieve crosslinking of collagen of jellyfish by the method of carbodiimides (edc/nhs). this method allowed us to increase the denaturation temperature to 33 ° c (vs. 28.9 ° c). the study of the interaction between human cells (mg63 and fibroblasts) and jellyfish collagen, demonstrated that they were able to adhere native or denatured jellyfish collagen. the immunostains of vinculin of mg63 cells and fibroblasts seeded on native or denatured jellyfish collagen, allowed us to characterize the focal adhesions of these two cell types. analysis of culture media of mg63 cells and fibroblasts by collagen zymography on native or denatured jellyfish collagen gels, allowed us to demonstrate that mmps of human origin, are able to degrade jellyfish collagen. we estimate in this case that the jellyfish collagen is a good substrate for the development of resorbable biomaterial