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European Cells and Materials Vol. 26. Suppl. 6, 2013 (page 1) ISSN 1473-2262 Sea-urchin inspired wet bioadhesives - a multidisciplinary quest towards biomimicry R Santos Biomedical and Oral Sciences Research Unit (UICOB), School of Dentistry of the University of Lisbon, Portugal INTRODUCTION: Adhesive mechanisms are underwater environment. More recently, beginning to be deciphered in some marine techniques such as proteomics and mass organisms that attach permanently with a cement spectrometry, demonstrated that tube feet are (mussels, barnacles) or use a viscous film to move unique mechano-sensory adhesive organs and and adhere (limpets). However, this is far from highlighted putative adhesive proteins. Work is in being true for sea urchins, that attach strongly but progress to identify the genes coding for these temporarily to the substratum. Sea urchins are adhesive proteins in order to produce recombinant exclusively benthic animals and to withstand analogues or partial adhesive motifs that can be hydrodynamic forces rely on hundreds of used to design new dental bioadhesives inspired on independent adhesive organs, their adoral tube millions of years of optimization through feet, specialized in locomotion and anchoring. biological selection. Tube feet consist of two functional units: a disc, which makes contact with the substratum, and a stem, which connects the disc to the test. Therefore, the tenacity with which an individual can anchor to a surface is determined by the strength of its numerous tube feet which, in turn, Fig. 1: Images of sea urchin tube feet attaching depends on the tensile strength of the stem and the underwater to the surface of a rock (A) and to adhesive power of the disc. The later is determined enamel and dentine in a human tooth slice both in by the chemical properties of an adhesive secretion (B). that is able to functionally assemble underwater and displace water, ions, and weakly bound DISCUSSION & CONCLUSIONS: Adhesives polyions, properties that are rarely achieved by produced by marine attaching organisms have synthetic adhesives. In addition, sea urchins also attracted attention as a paradigm of strong and possess an effective de-adhesive secretion that versatile underwater adhesion. Given the similarity rapidly releases the tube feet in order to begin a between the oral environment and the wave swept new attachment-detachment cycle. This de- seashore it is highly probable that water-efficient adhesive secretion is believed to contain enzymes, marine adhesives can be valuable sources of which catalytic activity promotes breaking of the inspiration to develop new dental adhesives. bonds established between the adhesive and the Although a number of improved dental adhesives disc, thus leaving a circle of adhesive material have become available commercially there are still (footprint) strongly attached to the substrate. a number of unsolved problems, reinforcing the METHODS & RESULTS: More than 10 years of need to develop new biomimetic adhesives based research combining morphological and on the knowledge of the molecular mechanisms biomechanical techniques have provided evidence underlying marine adhesion. that tube feet discs attach with a tenacity (adhesive ACKNOWLEDGEMENTS: Fundação para a force per unit area) similar to other marine and Ciência e Tecnologia for financing; COST Action commercial adhesives, being able to cope with TD0906 (http://www.cost-bioadhesives.org/) and different substrate chemistries and roughness, thus several colleagues and scientists, in particular Dr. ensuring strong adhesion irrespective of Patrick Flammang, for their valuable contribution substratum profile or chemistry. These findings over the years. demonstrated that sea urchins tube feet have evolved to resist hydrodynamic loads and deal with the unpredictability of the substratum, which has not yet been achieved by any synthetic (chemical-based) adhesive when applied in an http://www.ecmjournal.org European Cells and Materials Vol. 26. Suppl. 6, 2013 (page 2) ISSN 1473-2262 Evaluation of synthetic bone substitute engineered with amniotic epithelial cells in bone regeneration A Polimeni1, S Tetè2, EF Gherlone3 1Dept of Oral Science, University “La Sapienza”, Rome, Italy. 2Dept of Medical ,Oral and Biotechnological Science, University “G. d’Annunzio, Chieti, Italy. 3Dept of Dentistry, Vita Salute University, San Raffaele Hospital, Milan, Italy. INTRODUCTION: The objective of this study amnions of sheep slaughtered at approximately 3 was to in vitro and in vivo evaluate the efficacy, month of pregnancy. In order to achieve 3D safety and flexibility of an innovative cell-based scaffold loading, 10x106 labeled oAEC were tissue engineering approach for bone regeneration. inseminated on single blocks of synthetic bone Tissue engineering is an area of multidisciplinary substitute (∼1.4 cm3). Bilateral sinus augmentation research aimed at regenerating tissues and procedure are carried out with an extra-oral restoring organ functionality by means of the approach. Animals were euthanatized 45 and 90 transplant of cells and tissues developed in vitro or days after surgery and block section were obtained. by stimulating cellular growth in a synthetic Sinus explants were fixed in 4% paraformaldehyde matrix. The generated vital tissue should ideally be solution and analyzed with the micro immunologically, functionally, structurally and Computerized Tomography (micro-CT) technique mechanically identical to native tissue. In recent in order to evaluate the biomaterial integration into years, the efforts in this area focused on the the host tissue. The fixed explants were also characterization of the regenerative properties of processed for histological and immunohistological different sources of stem cells by studying analyses in order to detect the progression of embryonic and adult stem cells, but both cell types healing at different time. exhibit critical elements that have, until now, RESULTS: The results of this study suggest that limited their clinical application. For this reason it scaffold integration and bone deposition are was necessary to find an alternative source of cells positively influenced by the oAEC loading. for regenerative purpose. Amniotic derived cells Maxillary sinuses treated with the engineered (ADSs) are used in this study. ADSs are an biocomplexes displayed a limited inflammatory intermediate stage between embryonic stem cells response and an accelerated process of and lineage-restricted adult progenitor cells. Their angiogenesis. high proliferation rate together with their differentiation potential into cells of all 3 DISCUSSION & CONCLUSIONS: Transplanted embryonic germ layers (ectoderm, endoderm and oAEC were able to significantly increase the mesoderm) are important advantages over most of quantity of newly deposited bone tissue, as well as the known adult stem cell sources. Our research to stimulate a widespread process of angiogenesis includes the selection of a stem cell niche with and osteogenesis, which represent crucial events characteristics suitable for tissue regeneration, that successfully guiding tissue. In addition, oAEC is the amniotic epithelium, the development of a seem to be also able to directly participate in the suitable synthetic tridimensional scaffolds in order process of bone deposition, as suggested by the to build a biocomplex with stem cells, and the presence of themselves entrapped within the newly evaluation of the osteo-regenerative potential of deposited bone matrix. the obtained biocomplex for pre-clinical REFERENCES: 1 B. Stevens, Y. Yang, A. application. Mohandas, et al (2008) J Biomed Mater Res B. METHODS: Novel cell constructs, made by stem 85:573. 2 S. Hollister (2005) 4:518. 3 U. Deisinger, cells and synthetic tridimensional scaffolds S. Hamisch, M. Schumacher, et al (2008) Key (hydroxiapatite and beta-tricalcium phosphate in a Engineering Materials 361:915. 30/70 ratio or mineralized, solvent-dehydrated ACKNOWLEDGEMENTS: This study was bone allografts), were prepared for in vitro and in partly funded by PRIN 20102ZLNJ5. vivo tests, through the direct rapid prototyping technique dispense-plotting. For preclinical setting, sheep was chosen as animal model, because of its high translational value. The oAEC (ovine amniotic epithelial cells) were collected from http://www.ecmjournal.org European Cells and Materials Vol. 26. Suppl. 6, 2013 (page 3) ISSN 1473-2262 An overview of biomimetic approaches for biomedical adhesives development P Flammang, M Demeuldre, E Hennebert Biology of Marine Organisms and Biomimetics, Research Institute for Biosciences, University of Mons, Belgium from marine organisms, research into the INTRODUCTION: Many natural bioadhesives production of bio-inspired adhesives has perform in ways that man-made products simply developed in two different directions: the cannot match. Marine adhesives, for instance, production of recombinant adhesive proteins and work most effectively underwater and are the incorporation of functional adhesive motifs, universal in their performance to substrates of mostly DOPA, to chemically synthesized polymers varying composition and structure. They could (Fig. 1). Other remarkable properties of natural therefore potentially inspire novel, superior cements, such as the capacity to form complex adhesives for a large range of biomedical coacervates, to self-assemble and to cross-link, applications. Indeed, similarities exist between the have been mimicked for the underwater delivery marine and human physiologic