Eva Jablonská

Structure, mechanical characteristics and in vitro degradation, cytotoxicity, genotoxicity and mutagenicity of novel biodegradable Zn-Mg alloys.

Zn-(0-1.6)Mg (in wt.%) alloys were prepared by hot extrusion at 300 °C. The structure, mechanical properties and in vitro biocompatibility of the alloys were investigated. The hot-extruded magnesium-based WE43 alloy was used as a control. Mechanical properties were evaluated by hardness, compressive and tensile testing. The cytotoxicity, genotoxicity (comet assay) and mutagenicity (Ames test) of the alloy extracts and ZnCl2 solutions were evaluated with the use of murine fibroblasts L929 and human osteosarcoma cell line U-2 OS. The microstructure of the Zn alloys consisted of recrystallized Zn grains of 12 μm in size and fine Mg2Zn11 particles arranged parallel to the hot extrusion direction. Mechanical tests revealed that the hardness and strength increased with increasing Mg concentration. The Zn-0.8 Mg alloys showed the best combination of tensile mechanical properties (tensile yield strength of 203 MPa, ultimate tensile strength of 301 MPa and elongation of 15%). At higher Mg concentrations the plasticity of Zn-Mg alloys was deteriorated. Cytotoxicity tests with alloy extracts and ZnCl2 solutions proved the maximum safe Zn(2+) concentrations of 120 μM and 80 μM for the U-2 OS and L929 cell lines, respectively. Ames test with extracts of alloys indicated that the extracts were not mutagenic. The comet assay demonstrated that 1-day extracts of alloys were not genotoxic for U-2 OS and L929 cell lines after 1-day incubation.

Highly porous, low elastic modulus 316L stainless steel scaffold prepared by selective laser melting

Recently, porous metallic materials have been extensively studied as candidates for use in the fabrication of scaffolds and augmentations to repair trabecular bone defects, e.g. in surroundings of joint replacements. Fabricating these complex structures by using common approaches (e.g., casting and machining) is very challenging. Therefore, rapid prototyping techniques, such as selective laser melting (SLM), have been investigated for these applications. In this study, we characterized a highly porous (87 vol.%) 316L stainless steel scaffold prepared by SLM. 316L steel was chosen because it presents a biomaterial still widely used for fabrication of joint replacements and, from the practical point of view, use of the same material for fabrication of an augmentation and a joint replacement is beneficial for corrosion prevention. The results are compared to the reported properties of two representative nonporous 316L stainless steels prepared either by SLM or casting and subsequent hot forging. The microstructural and mechanical properties and the surface chemical composition and interaction with the cells were investigated. The studied material exhibited mechanical properties that were similar to those of trabecular bone (compressive modulus of elasticity ~0.15GPa, compressive yield strength ~3MPa) and cytocompatibility after one day that was similar to that of wrought 316L stainless steel, which is a commonly used biomaterial. Based on the obtained results, SLM is a suitable method for the fabrication of porous 316L stainless steel scaffolds with highly porous structures.

Microstructural, mechanical, corrosion and cytotoxicity characterization of the hot forged FeMn30(wt.%) alloy.

An interest in biodegradable metallic materials has been increasing in the last two decades. Besides magnesium based materials, iron-manganese alloys have been considered as possible candidates for fabrication of biodegradable stents and orthopedic implants. In this study, we prepared a hot forged FeMn30 (wt.%) alloy and investigated its microstructural, mechanical and corrosion characteristics as well as cytotoxicity towards mouse L 929 fibroblasts. The obtained results were compared with those of iron. The FeMn30 alloy was composed of antiferromagnetic γ-austenite and ε-martensite phases and possessed better mechanical properties than iron and even that of 316 L steel. The potentiodynamic measurements in simulated body fluids showed that alloying with manganese lowered the free corrosion potential and enhanced the corrosion rate, compared to iron. On the other hand, the corrosion rate of FeMn30 obtained by a semi-static immersion test was significantly lower than that of iron, most likely due to a higher degree of alkalization in sample surrounding. The presence of manganese in the alloy slightly enhanced toxicity towards the L 929 cells; however, the toxicity did not exceed the allowed limit and FeMn30 alloy fulfilled the requirements of the ISO 10993-5 standard.

A novel high-strength and highly corrosive biodegradable Fe-Pd alloy: Structural, mechanical and in vitro corrosion and cytotoxicity study

Recently, iron-based materials have been considered as candidates for the fabrication of biodegradable load-bearing implants. Alloying with palladium has been found to be a suitable approach to enhance the insufficient corrosion rate of iron-based alloys. In this work, we have extensively compared the microstructure, the mechanical and corrosion properties, and the cytotoxicity of an FePd2 (wt%) alloy prepared by three different routes - casting, mechanical alloying and spark plasma sintering (SPS), and mechanical alloying and the space holder technique (SHT). The properties of the FePd2 (wt%) were compared with pure Fe prepared in the same processes. The preparation route significantly influenced the material properties. Materials prepared by SPS possessed the highest values of mechanical properties (CYS~750-850MPa) and higher corrosion rates than the casted materials. Materials prepared by SHT contained approximately 60% porosity; therefore, their mechanical properties reached the lowest values, and they had the highest corrosion rates, approximately 0.7-1.2mm/a. Highly porous FePd2 was tested in vitro according to the ISO 10993-5 standard using L929 cells, and two-fold diluted extracts showed acceptable cytocompatibility. In general, alloying with Pd enhanced both mechanical properties and corrosion rates and did not decrease the cytocompatibility of the studied materials.

Influence of surface pre-treatment on the cytocompatibility of a novel biodegradable ZnMg alloy.

Degradable zinc-based alloys with an appropriate corrosion rate are promising materials for the preparation of temporary orthopaedic implants. Previously, we prepared and characterised a novel Zn1.5Mg alloy. This paper is focused on the characterisation of this alloy after a surface pre-treatment, which should mimic processes occurring in vivo. The samples of the Zn1.5Mg alloy were immersed in a simulated body fluid (SBF) at 37°C for 14days in order to form a protective layer of corrosion products. Thereafter, these samples were used for the corrosion rate determination, an indirect in vitro cytotoxicity test, as well as for a direct contact test and were compared with the non-treated samples. The protective layer was characterized by SEM and its chemical composition was determined by EDS and XPS analysis. The corrosion rate was significantly decreased after the pre-incubation. The protective layer of corrosion products was rich in Ca and P. The pre-incubated samples exhibited increased cytocompatibility in the indirect test (metabolic activity of L929 cells was above 70%) and we also observed osteoblast-like cell growth directly on the samples during the contact tests. Thus, the pre-incubation in SBF leading to improved cytocompatibility could represent more appropriate model to in vivo testing.

Effect of equal channel angular pressing on in vitro degradation of LAE442 magnesium alloy

Effect of processing by equal channel angular pressing (ECAP) on the degradation behaviour of extruded LAE442 magnesium alloy was investigated in a 0.1M NaCl solution, Kirklands biocorrosion medium (KBM) and Minimum Essential Medium (MEM), both with and without 10% of foetal bovine serum (FBS). Uniform degradation of as extruded and ECAP processed samples in NaCl solution was observed, nevertheless higher corrosion resistance was found in the latter material. The increase of corrosion resistance due to ECAP was observed also after 14-days immersion in all media used. Higher compactness of the corrosion layer formed on the samples after ECAP was responsible for the observed decrease of corrosion resistance, which was proven by scanning electron microscope investigation. Lower corrosion rate in media with FBS was observed and was explained by additional effect of protein incorporation on the corrosion layer stability. A cytotoxicity test using L929 cells was carried out to investigate possible effect of processing on the cell viability. Sufficient cytocompatibility of the extruded samples was observed with no adverse effects of the subsequent ECAP processing. In conclusion, this in vitro study proved that the degradation behaviour of the LAE442 alloy could be improved by subsequent ECAP processing and this material is a good candidate for future in vivo investigation.

Novel Approach in the Use of Plasma Spray: Preparation of Bulk Titanium for Bone Augmentations

Thermal plasma spray is a common, well-established technology used in various application fields. Nevertheless, in our work, this technology was employed in a completely new way; for the preparation of bulk titanium. The aim was to produce titanium with properties similar to human bone to be used for bone augmentations. Titanium rods sprayed on a thin substrate wire exerted a porosity of about 15%, which yielded a significant decrease of Young′s modulus to the bone range and provided rugged topography for enhanced biological fixation. For the first verification of the suitability of the selected approach, tests of the mechanical properties in terms of compression, bending, and impact were carried out, the surface was characterized, and its compatibility with bone cells was studied. While preserving a high enough compressive strength of 628 MPa, the elastic modulus reached 11.6 GPa, thus preventing a stress-shielding effect, a generally known problem of implantable metals. U-2 OS and Saos-2 cells derived from bone osteosarcoma grown on the plasma-sprayed surface showed good viability.

Titania sol-gel coatings containing silver on newly developed TiSi alloys and their antibacterial effect

New materials with appropriate mechanical properties and an antibacterial effect are constantly being sought for orthopedic and dental applications. The aim of this study was to investigate newly developed TiSi alloys coated with titania sol-gel containing silver. Titanium alloys with 5 or 10wt% of silicon were prepared by vacuum arc remelting and dip-coated with titania sol containing either AgNO3 or Ag3PO4 in two concentrations. The size and distribution of the particles in the layer were evaluated, as well as layer compactness (SEM). The antibacterial effect (against E. coli and S. epidermidis) and cytotoxicity (towards L929 and U-2 OS cell lines) of these materials were then tested. Despite cracking of the coatings after firing, the coatings demonstrated very good antibacterial effects against both E. coli and S. epidermidis after 24h of interaction. None of the tested materials were toxic to both cell lines. Collectively, our results suggest that these materials are promising candidates for orthopedic applications.

Corrosion behaviour and cell interaction of Ti-6Al-4V alloy prepared by two techniques of 3D printing

3D printing seems to be the technology of the future for the preparation of metallic implants. For such applications, corrosion behaviour is pivotal. However, little is published on this topic and with inconsistent results. Therefore, we carried out a complex study in which we compared two techniques of the 3D printing technology - selective laser melting and electron beam melting. The corrosion behaviour was studied in physiological solution by standard electrochemical techniques and susceptibility to localised corrosion was estimated too. All samples showed typical passive behaviour. Localised corrosion was shown to be possible on the original as-printed surfaces. Corrosion experiments were repeated tree times. To reveal possible negative effects of 3D printing on cytocompatibility, direct in vitro tests were performed with U-2 OS cells. The cells showed good viability and proliferation, but their growth was impeded by surface unevenness. Our results suggest that both techniques are suitable for implants production. Statistical evaluation was performed by ANOVA followed by Tukeys test.

Titania sol-gel coatings with silver on non-porous titanium and titanium alloys

The objective of the work was to prepare and characterize titania sol-gel coatings on non-porous titanium and newly developed titanium alloys. Basic titania sol contained two forms of silver. Titania sol without silver was used as a reference sample. Coatings were prepared by dip-coating technique during stirring and fired. Coatings after firing were characterized by scanning electron microscopy. All titania coatings were measured to determine their adhesive and bactericidal properties. Adhesion of the coatings to the substrate was measured by tape test. Gram-negative bacteria E. coli was used for the bactericidal test. Coated substrates were immersed into suspension of E. coli in physiological solution for 24 hours. The in vitro cytotoxicity test was performed after one day. The bactericidal effect without toxicity was confirmed for selected coatings.