Mónica A. Fernández Lorenzo de Mele

Critical discussion of the results from different corrosion studies of Mg and Mg alloys for biomaterial applications.

The aim of this work was to collect and compare data from different published reports which focused on the description of the influence of different electrochemical setups for the assessment of magnesium corrosion. Based on this, a comparison with our own results, obtained for LAE 442 and AZ 31, was made and discussed. As the collection of data has shown, the reported inconsistencies between in vivo and electrochemical data depend greatly on the electrochemical medium used, on the alloy composition and on surface preparation. Nevertheless, these differences also exist when comparing different in vitro results using different methodologies and even different Mg alloys, and need therefore to be discussed more thoroughly in the future. The simulation of transport conditions of the in vivo interface should become a focus of research interest in order to gain a better understanding of the influence of connecting processes on the degradation of the biomaterials.

Degradation of magnesium and its alloys: Dependence on the composition of the synthetic biological media

Magnesium and its alloys are highly degradable metals that are potentially useful as biomaterials, especially in orthopaedic and cardiovascular applications. However, the in vivo corrosion has proved to be too high. Because of the complexity of in vivo conditions, a careful study of the corrosion of magnesium in synthetic solutions that simulate the in vivo environment is necessary as a first approach to predict the actual in vivo situation. The aim of this work was to evaluate the influence of the electrolyte composition on the corrosion behavior of magnesium and two Mg-alloys in synthetic biological media. Pure magnesium and its alloys (AZ31 and LAE442) were employed in the experiments. Electrochemical potentiodynamic polarization curves were recorded in sodium chloride and PBS electrolytes with different chloride ion and albumin concentration. Optical and SEM observations complemented by EDX analysis were made. The results showed that magnesium corrosion is localized in chloride- and albumin-containing buffer solutions. They also showed that the chloride concentration and the presence of buffer and protein strongly affect the electrochemical behavior of magnesium and magnesium alloys.

Magnesium and its alloys as degradable biomaterials: corrosion studies using potentiodynamic and EIS electrochemical techniques

Magnesium is potentially useful for orthopaedic and cardiovascular applications. However, the corrosion rate of this metal is so high that its degradation occurs before the end of the healing process. In industrial media the behaviour of several magnesium alloys have been probed to be better than magnesium performance. However, the information related to their corrosion behaviour in biological media is insufficient. The aim of this work is to study the influence of the components of organic fluids on the corrosion behaviour of Mg and AZ31 and LAE442 alloys using potentiodynamic, potentiostatic and EIS techniques. Results showed localized attack in chloride containing media. The breakdown potential decreased when chloride concentration increased. The potential range of the passivation region was extended in the presence of albumin. EIS measurements showed that the corrosion behaviour of the AZ31 was very different from that of LAE442 alloy in chloride solutions.

Metallic dental material biocompatibility in osteoblastlike cells

Ions released from metallic dental materials used in orthodontic appliances could induce undesirable effects on cells and tissues. This study evaluates the biocompatibility of two of the most labile components of metallic dental alloys on osteoblastlike cells. The influence of protein and ions on metal dissolution properties is also investigated using different electrolyte solutions. Morphological alterations, cell growth, and differentiation of osteoblasts were assessed after exposure to pure metals (Ag, Cu, Pd, Au) and Ni−Ti alloy and correlated with the kinetics of elements released into the culture media. Results showed that Cu and Ag were the most cytotoxic elements and the other metals were biocompatible with the osteoblasts. The parameters of biocompatibility were correlated with the levels of ions detected into the culture media. Metal ions induced cell death through early mitosis arrest, apoptotic phenomena, and necrotic processes. Voltammograms showed that anions and proteins interfered in the corrosion process. Fetal bovine serum (FBS) strongly affected the electrochemical process, decreasing the oxidation rate of the metals. In conclusion, copper and silver ions showed a time-dependent low biocompatibility, which correlated with the concentration of released ions. The dissolution of the metallic materials was dependent on the composition of the simulated biological media.

Influence of the nano-micro structure of the surface on bacterial adhesion

Biomaterials failures are frequently associated to the formation of bacterial biofilms on the surface. The aim of this work is to study the adhesion of non motile bacteria streptococci consortium and motile Pseudomonas fluorescens. Substrates with micro and nanopatterned topography were used. The influence of surface characteristics on bacterial adhesion was investigated using optical and epifluorescence microscopy, scanning electron microscopy (SEM) and atomic force microscopy (AFM). Results showed an important influence of the substratum nature. On microrough surfaces, initial bacterial adhesion was less significant than on smooth surfaces. In contrast, nanopatterned samples showed more bacterial attachment than the smooth control. It was also noted a remarkable difference in morphology, orientation and distribution of bacteria between the smooth and the nanostructured substrate. The results show the important effect of substratum nature and topography on bacterial adhesion which depended on the relation between roughness characteristics dimensions and bacterial size.

Submicron trenches reduce the Pseudomonas fluorescens colonization rate on solid surfaces.

Bacterial adhesion and spreading on biomaterials are considered key features of pathogenicity. Roughness and topography of the substrate have been reported to affect bacterial adhesion, but little is known about their effect on spreading. Submicron row and channel tuning with bacterial diameter (S2) were designed to test bacterial motility on these surfaces. Random nanometer-sized structures (S1) were used as controls. Optical microscopy and AFM were employed to detect biological and surface pattern details in the micro- and nanoscale, respectively. Results showed that motility strategies (flagella orientation, elongation, aggregation in rafts, formation of network structures, and development of a bacterial frontier) were affected by the presence of submicropatterns. Importantly, the rate of bacterial spreading on S2 was significantly reduced and influenced by the orientation of the submicropatterns. Consequently, submicroengineered substrates could be employed as a tool to downgrade bacterial colonization. Such patterns could impact on the design of proper engineered structures to control biofilm spreading on solid surfaces.

Does over-exposure to copper ions released from metallic copper induce cytotoxic and genotoxic effects on mammalian cells?

BACKGROUND A high dissolution of copper from intrauterine devices (IUDs) occurs during the first days after insertion. This work is focused on the assessment of the possible cyto- and genotoxic effects of different concentrations of copper ions released from metallic copper on mammalian cells in vitro. STUDY DESIGN Colorimetric tetrazolium/Trypan blue (TB) tests and Comet assay were used to evaluate potential cytotoxicity and genotoxicity, respectively, in Chinese hamster ovary cells (CHO-K1). RESULTS Reduction of mitochondrial activity by copper ions was observed for extracts at >or=7.42 mg/L concentrations, while TB exclusion test for plasma membrane integrity showed significant decrease in cell viability (close to 90%) for 10.85 mg/L concentration. Additionally, copper-induced DNA damage was detected for 5.67-7.42 mg/L concentration range. CONCLUSION Our results demonstrate cytotoxic and genotoxic effects of copper ions released from metallic copper on CHO-K1 cells and emphasize the importance of reducing the initial copper dissolution from IUD without affecting the contraceptive action.

Cytotoxicity of copper ions released from metal: variation with the exposure period and concentration gradients.

The aim of this work is to contribute to the elucidation of the cytotoxic process caused by the copper ions released from the biomaterials. Clonal cell lines UMR106 were used in the experiments. Copper ions were obtained from two different sources: copper salts and metal dissolution. Experiments carried out with constant ion concentrations (copper salts) were compared with those with concentrations that vary with time and location (dissolution of the metal). Present results and others previously reported could be interpreted through mathematical models that describe: (1) the variation of concentration of copper ions with time and location within a biofilm and (2) the variation of the killing rate with the concentration of the toxic ion and time. The large number of dead cells found near the copper sample with an average ion concentration below the toxic limit could be interpreted bearing in mind that these cells should be exposed to a local concentration higher than this limit. A logarithmic dependence between the number of cells and exposure time was found for nearly constant ion concentrations. Apparent discrepancies, observed when these results and those of different researchers were contrasted, could be explained considering the dissimilar experimental conditions such as the source of the ions and their local concentration at real time.

Organization of Pseudomonas fluorescens on Chemically Different Nano/Microstructured Surfaces

This paper describes bacterial organization on nano/micropatterned surfaces with different chemical properties, which show different interactions with the biological systems (inert, biocompatible, and bactericide). These surfaces were prepared by molding techniques and exposed to Pseudomonas fluorescens (P. fluorescens) cultures. Results from atomic force microscopy and optical imaging demonstrate that the structure of P. fluorescens aggregates is strongly dependent on the surface topography while there is no clear linking with the physical-chemical surface properties (charge and contact angle) of the substrate immersed in abiotic culture media. We observe that regardless of the material when the surface pattern matches the bacterial size, bacterial assemblages involved in surface colonization are disorganized. The fact there is not a relationship between surface chemistry and bacterial organization can be explained by the coverage of the surfaces by adsorbed organic species coming from the culture medium. Viability assays indicate that copper behaves as a toxic substrate despite the presence of adsorbed molecules. The combination of surface traps and biocidal activity could act synergistically as a suitable strategy to limit bacterial spreading on implant materials.

Chlorhexidine delivery system from titanium/polybenzyl acrylate coating: Evaluation of cytotoxicity and early bacterial adhesion

OBJECTIVES The formation of biofilms on titanium dental implants is one of the main causes of failure of these devices. Streptococci are considered early colonizers that alter local environment favouring growing conditions for other colonizers. Chlorhexidine (CHX) is so far the most effective antimicrobial treatment against a wide variety of Gram-positive and Gram-negative organisms as well as fungi. This study was designed to develop a CHX delivery system appropriate for healing caps and abutments, with suitable drug release rate, effective as antimicrobial agent, and free of cytotoxic effects. METHODS Polybenzyl acrylate (PBA) coatings with and without CHX (Ti/PBA and Ti/PBA-CHX, respectively) and different drug loads (0.35, 0.70, and 1.40%, w/w) were assayed. The cytotoxic effect of CHX released from the different substrates on UMR106 cells was tested by alkaline phosphatase specific activity (ALP), and microscopic evaluation of the cells. Non-cytotoxic drug load (0.35%, w/w) was selected to evaluate the antimicrobial effectiveness of the system using a microbial consortium of Streptococcus species. RESULTS The kinetic profile of CHX delivered by Ti/PBA-CHX showed an initial fast release rate followed by a monotonic increase of delivered mass over 48 h. The number of attached bacteria decreased in the following order: Ti>Ti/PBA>Ti/PBA-0.35. CONCLUSIONS PBA-0.35 coating is effective to inhibit the adhesion of early colonizers on Ti without any cytotoxic effect on UMR-106 cells.