Gisela M. Luz

Biomimetic design of materials and biomaterials inspired by the structure of nacre

The micro-architecture of nacre (mother of pearl) has been classically illustrated as a ‘brick-and-mortar’ arrangement. It is clear now that hierarchical organization and other structural features play an important role in the amazing mechanical properties of this natural nanocomposite. The more important structural characteristics and mechanical properties of nacre are exposed as a base that has inspired scientists and engineers to develop biomimetic strategies that could be useful in areas such as materials science, biomaterials development and nanotechnology. A strong emphasis is given on the latest advances on the synthetic design and production of nacre-inspired materials and coatings, in particular to be used in biomedical applications.

Chitosan/bioactive glass nanoparticle composite membranes for periodontal regeneration

Barrier membranes are used in periodontal applications with the aim of supporting periodontal regeneration by physically blocking migration of epithelial cells. The present work proposes a combination of chitosan (CHT) with bioactive glass nanoparticles (BG-NPs) in order to produce a novel guided tissue and bone regeneration membrane, fabricated by solvent casting. The CHT/BG-NP nanocomposite membranes are characterized in terms of water uptake, in mechanical tests, under simulated physiological conditions and in in vitro bioactivity tests. The addition of BG-NPs to CHT membranes decreased the mechanical potential of these membranes, but on the other hand the bioactivity improved. The membranes containing the BG-NPs induced the precipitation of bone-like apatite in simulated body fluid (SBF). Biological tests were carried out using human periodontal ligament cells and human bone marrow stromal cells. CHT/BG-NP composite membranes promoted cell metabolic activity and mineralization. The results indicate that the CHT/BG-NP composite membrane could potentially be used as a temporary guided tissue regeneration membrane in periodontal regeneration, with the possibility to induce bone regeneration.

Nanostructured Polymeric Coatings Based on Chitosan and Dopamine‐Modified Hyaluronic Acid for Biomedical Applications

In a marine environment, specific proteins are secreted by mussels and used as a bioglue to stick to a surface. These mussel proteins present an unusual amino acid 3,4-dihydroxyphenylalanine (known as DOPA). The outstanding adhesive properties of these materials in the sea harsh conditions have been attributed to the presence of the catechol groups present in DOPA. Inspired by the structure and composition of these adhesive proteins, dopamine-modified hyaluronic acid (HA-DN) prepared by carbodiimide chemistry is used to form thin and surface-adherent dopamine films. This conjugate was characterized by distinct techniques, such as nuclear magnetic resonance and ultraviolet spectrophotometry. Multilayer films are developed based on chitosan and HA-DN to form polymeric coatings using the layer-by-layer methodology. The nanostructured films formation is monitored by quartz crystal microbalance. The film surface is characterized by atomic force microscopy and scanning electron microscopy. Water contact angle measurements are also conducted. The adhesion properties are analyzed showing that the nanostructured films with dopamine promote an improved adhesion. In vitro tests show an enhanced cell adhesion, proliferation and viability for the biomimetic films with catechol groups, demonstrating their potential to be used in distinct biomedical applications.

Chitosan/bioactive glass nanoparticles composites for biomedical applications

Nanocomposite films based on a chitosan blend with bioactive glass nanoparticles (BG-NPs) with different formulations, namely SiO(2):CaO:P(2)O(5)(mol.%) = 55:40:5 and SiO(2):CaO:P(2)O(5):MgO(mol.%) = 64:26:5:5 were produced in order to develop systems with applicability in guided tissue regeneration. The zeta (ζ)-potential of the BG-NPs containing magnesium was found to be lower than the other formulation and the corresponding composite with chitosan was the most hydrophilic. The bioactive character of the biomaterials was also assessed in vitro by immersion of the materials in simulated body fluid, followed by scanning electron microscopy (SEM) and energy-dispersive x-ray spectroscopy evaluations. SaOs-2 osteoblastic-like cells were seeded on the different nanocomposites and their behavior was followed by SEM observations, cytotoxicity assessments, DNA quantification and alkaline phosphatase analysis. The introduction of the inorganic component in the chitosan matrix had a positive effect on the biological response of the membranes. The developed nanocomposite films are potential candidates for regenerating damaged bone tissue and could be useful in orthopedic and maxillo-facial applications.

Micropatterning of bioactive glass nanoparticles on chitosan membranes for spatial controlled biomineralization

Bioactive glass nanoparticles (BG-NPs) capable of inducing apatite precipitation upon immersion in simulated body fluid (SBF) were patterned on free-standing chitosan membranes by microcontact printing using a poly(dimethylsiloxane) (PDMS) stamp inked in a BG-NPs pad. Formation of the patterns was characterized by scanning electron microscopy (SEM). Mineralization of the bioactive glass patterns was induced in vitro by soaking the samples in SBF over different time points up to 7 days. The confined apatite deposition in the patterned regions with diameters of 50 μm was confirmed by Fourier-transformed infrared spectroscopy (FTIR), energy-dispersive X-ray (EDX) analysis, and SEM. In vitro tests confirmed the preferential attachment and proliferation of L929 cells to the areas printed with BG-NPs of the membranes. This approach permits one to spatially control the properties of biomaterials at the microlevel and could be potentially used in guided tissue regeneration for skin, vascular, articular, and bone tissue engineering and in cellular cocultures or to develop substrates able to confine cells in regions with controlled geometry at the cells length scale.

Adhesive Bioactive Coatings Inspired by Sea Life

Inspired by nature, in particular by the marine mussels adhesive proteins (MAPs) and by the tough brick-and-mortar nacre-like structure, novel multilayered films are prepared in the present work. Organic-inorganic multilayered films, with an architecture similar to nacre based on bioactive glass nanoparticles (BG), chitosan, and hyaluronic acid modified with catechol groups, which are the main components responsible for the outstanding adhesion in MAPs, are developed for the first time. The biomimetic conjugate is prepared by carbodiimide chemistry and analyzed by ultraviolet-visible spectrophotometry. The buildup of the multilayered films is monitored with a quartz crystal microbalance with dissipation monitoring, and their topography is characterized by atomic force microscopy. The mechanical properties reveal that the films containing catechol groups and BG present an enhanced adhesion. Moreover, the bioactivity of the films upon immersion in a simulated body fluid solution is evaluated by scanning electron microscopy coupled with energy dispersive X-ray spectroscopy, Fourier transform infrared spectroscopy, and X-ray diffraction. It was found that the constructed films promote the formation of bonelike apatite in vitro. Such multifunctional mussel inspired LbL films, which combine enhanced adhesion and bioactivity, could be potentially used as coatings of a variety of implants for orthopedic applications.

Poly(ɛ-caprolactone) Electrospun Scaffolds Filled with Nanoparticles. Production and Optimization According to Taguchi's Methodology

Polycaprolactone (PCL) scaffolds were produced by electrospinning. Polymeric solutions in a mix of dichloromethane (DCM) and dimethylformamide were electrospun to form fibers in the sub-micron range. Physical properties of the PCL solutions were characterized with respect to density, viscosity, conductivity and surface tension. Processing was optimized following Taguchis methodology to select the set of processing parameters that resulted in producing fibers with the smallest diameters, minimum number of defects and with the narrowest distribution of fiber diameter. Morphology of electrospun fibers was qualitatively and quantitatively analyzed for the different sets of processing parameters. The optimum conditions found to electrospun PCL were used to process PCL solutions containing nanoparticles of hydroxyapatite (HA) or bioactive glass (BG). Bioactivity of nanocomposite electrospun membranes in simulated body fluid (SBF) was analyzed and biological response was tested by assessing proliferation and viability of MT3C3-E1 preosteoblasts cultured on PCL and its nanocomposite membranes.

A nanotectonics approach to produce hierarchically organized bioactive glass nanoparticles-based macrospheres

Bioactive particles have been widely used in a series of biomedical applications due to their ability to promote bone-bonding and elicit favorable biological responses in therapies associated with the replacement and regeneration of mineralized tissues. In this work hierarchical architectures are prepared by an innovative methodology using SiO(2)-CaO sol-gel based nanoparticles. Inspired by colloidal crystals, spherical aggregates were formed on biomimetic superhydrophobic surfaces using bioactive glass nanoparticles (BG-NPs) able to promote bone regeneration. A highly ordered organization, a common feature of mineralized structures in Nature, was achieved at both nano- and microlevels, being the crystallization degree of the structures controlled by the evaporation rates taking place at room temperature (RT) or at 4 °C. The crystallization degree of the structures influenced the Ca/P ratio of the apatitic film formed at their surface, after 7 days of immersion in SBF. This allows the regulation of bioactive properties and the ability to release potential additives that could be also incorporated in such particles with a high efficiency. Such a versatile method to produce bioactive particles with controlled size and internal structure could open new possibilities in designing new spherical devices for orthopaedic applications, including tissue engineering.

Membranes of poly(dl-lactic acid)/Bioglass® with asymmetric bioactivity for biomedical applications

In guided tissue/bone regeneration, membranes are used as barriers to prevent the faster growing soft tissue cells from entering the defect space and to regenerate periodontal ligament, cementum, and bone. The two sides of the membranes are in contact with distinct biological environments in which one faces a region in which osteointegration should be ideally promoted. Biocompatible and biodegradable composite membranes were produced by combining poly(d,l-lactic acid) and Bioglass® particles featuring an asymmetric bioactivity and a good integration between the polymeric and inorganic fractions. The asymmetric distribution of the osteoconductive particles was produced during the processing of the membrane using a solvent casting methodology. Only the inorganic-rich face promoted the deposition of bone-like apatite after immersing the composite membrane in simulated body fluid for 2 days. The mechanical properties of the membranes were evaluated using dynamic mechanical analysis by analyzing the viscoelastic properties and the glass transition of the samples in both dry and wet states. A clear plasticization effect of water was detected, but the composite membranes were found to be stiffer, at 37ºC, compared with the pure polymer. SaOs-2 cells attached on both the surfaces and proliferated after 7 days in culture.

Development of a bioactive glass-polymer composite for wound healing applications

This study reports the production and characterization of a composite material for wound healing applications. A bioactive glass obtained by sol-gel process and doped with two different metal ions was investigated. Silver (Ag) and cobalt (Co) were chosen due to their antibacterial and angiogenic properties, respectively, very beneficial in the wound healing process. Poly(ε-caprolactone) (PCL) fibers were produced by electrospinning (ES) from a polymeric solution using acetone as a solvent. After optimization of the ES parameters, two main suspensions were prepared, namely: PCL containing bioactive glass nanoparticles (BG-NP) and PCL with Ag2O and CoO doped BG-NP (DP BG-NP), which were processed with different concentrations of BG-NP (0.25%, 0.5% and 0.75wt%). The composite membranes were characterized in terms of morphology, fiber diameter, weight loss, mineralization potential and mechanical performance.