Combinatorial analysis of marine based biomaterials: high-throughput analysis of the effect of nanostructured multilayers on cell behaviour

In a marine environment, specific proteins are secreted by mussels and used as a bioglue to stick to a surface allowing generate irreversible bonding. Adhesive secreted proteins of mussels present an unusual amino acid 3,4-dihydroxyphenylalanine (DOPA). Inspired by the struc- ture and properties of mussel adhesive proteins, layer-by-layer (LbL) coatings based on polymers that contain catechol groups were devel- oped. We used dopamine-modified hyaluronic acid (HA-DN) prepared by carbodiimide chemistry to form thin and surface-adherent dopamine films. The multilayer films were developed by electrostatic interactions using chitosan (CHT) as polycation and HA-DN as polyanion. Multilay- ers films of CHT and HA were used as control. The formation of these films was investigated in-situ by quartz crystal microbalance with dissi- pation monitoring (QCM-D). Afterwards, many combinations of the marine inspired biomaterials were analysed in a high-throughput (HTS) way. Such multilayers were constructed and individually dis- posed on isolated transparent spots, patterned onto biomimetic super- hydrophobic substrates. The adhesion properties of the coatings in the chips were also analyzed. In vitro tests using two distinct cell sources were carried out to evaluate the biological performance of the different combinations of multilayers that could be useful in different biomedical applications, including tissue engineering.