Oceans are a vast source of natural substances. In them, we find various compounds with wide biotechnological and biomedical applicabilities. The exploitation of the sea as a renewable source of biocompounds can have a positive impact on the development of new systems and devices for biomedical applications. Marine polysaccharides are among the most abundant materials in the seas, which contributes to a decrease of the extraction costs, besides their solubility behavior in aqueous solvents and extraction media, and their interaction with other biocompounds. Polysaccharides such as alginate, carrageenan and fucoidan can be extracted from algae, whereas chitosan and hyaluronan can be obtained from animal sources. Most marine polysaccharides have important biological properties such as biocompatibility, biodegradability, and anti-inflammatory activity, as well as adhesive and antimicrobial actions. Moreover, they can be modified in order to allow processing them into various shapes and sizes and may exhibit response dependence to external stimuli, such as pH and temperature. Due to these properties, these biomaterials have been studied as raw material for the construction of carrier devices for drugs, including particles, capsules and hydrogels. The devices are designed to achieve a controlled release of therapeutic agents in an attempt to fight against serious diseases, and to be used in advanced therapies, such as gene delivery or regenerative medicine.

Layer-by-layer (LbL) was first introduced as a surface modification technique based on the sequential spontaneous adsorption of at least two distinct materials onto planar substrates. In the last two decades, this technique has been expanded to the coating of more convoluted geometries with high levels of tailored functionalization or with structural purposes. In this review, the potential uses of LbL films in biomedical engineering based mainly on the assembly of polyelectrolytes are reviewed. Examples of recent developments are provided, from the modification of substrates to improve their biointegration or to add specialized properties, to the three-dimensional extrapolation of this technique to more complex structures for cell seeding, drug delivery devices, biosensors and customizable microreactors. Future strategies and opportunities are compared with current medical and laboratorial methodologies. Through them, it is expected that LbL will contribute greatly to the development of new functional devices with high perspectives of return for the administration of active agents, supports for cells in regenerative medicine and tissue engineering, biosensing and construction of microtissues and disease models in the laboratory.

Nanostructured three-dimensional constructs combining layer-by-layer technology (LbL) and template leaching were processed and evaluated as possible support structures for cartilage tissue engineering. Multilayered constructs were formed by depositing the polyelectrolytes chitosan (CHT) and chondroitin sulphate (CS) on either bidimensional glass surfaces or 3D packet of paraffin spheres. 2D CHT/CS multi-layered constructs proved to support the attachment and proliferation of bovine chondrocytes (BCH). The technology was transposed to 3D level and CHT/CS multi-layered hierarchical scaffolds were retrieved after paraffin leaching. The obtained nanostructured 3D constructs had a high porosity and water uptake capacity of about 300%. Dynamical mechanical analysis (DMA) showed the viscoelastic nature of the scaffolds. Cellular tests were performed with the culture of BCH and multipotent bone marrow derived stromal cells (hMSCs) up to 21 days in chondrogenic differentiation media. Together with scanning electronic microscopy analysis, viability tests and DNA quantification, our results clearly showed that cells attached, proliferated and were metabolically active over the entire scaffold. Cartilaginous extracellular matrix (ECM) formation was further assessed and results showed that GAG secretion occurred indicating the maintenance of the chondrogenic phenotype and the chondrogenic differentiation of hMSCs.

Medical adhesives and sealants often require that long-term adhesiveness is achieved. In this work, nanostructured coatings consisting of chitosan and the adhesive bacterial exopolysaccharide levan are fabricated using layer-by-layer (LbL) assembly. Taking advantage of the electrostatic self-assembly mechanism of LbL, the charges of both chitosan and a phosphonate-derivatized levan (Ph-levan) are measured and the feasibility of constructing hybrid films is monitored and confirmed using a quartz crystal microbalance with dissipation monitoring (QCM-D). The adhesive properties between two identical bonded films with a total of 100 layers are compared to control films in which Ph-levan is replaced by alginate, revealing that the detachment force of the former is about 3 times higher than the control. Scanning electron microscopy of the films surface shows that the surface of Ph-levan films is smooth and homogeneous. Cell adhesion tests were conducted using a L929 cell line. Early cell adhesion is significantly higher in chitosan/Ph-levan films when compared to chitosan/alginate controls. These findings establish levan derivatives as bioinspired ingredients for conceiving medical adhesive devices that allow achieving enhanced mechanical and biological performance.