In Vivo Simulation of Magnesium Degradability Using a New Fluid Dynamic Bench Testing Approach.

The degradation rate of magnesium (Mg) alloys is a key parameter to develop Mg-based biomaterials and ensure in vivo-mechanical stability as well as to minimize hydrogen gas production, which otherwise can lead to adverse effects in clinical applications. However, in vitro and in vivo results of the same material often differ largely. In the present study, a dynamic test bench with several single bioreactor cells was constructed to measure the volume of hydrogen gas which evolves during magnesium degradation to indicate the degradation rate in vivo. Degradation medium comparable with human blood plasma was used to simulate body fluids. The media was pumped through the different bioreactor cells under a constant flow rate and 37 °C to simulate physiological conditions. A total of three different Mg groups were successively tested: Mg WE43, and two different WE43 plasma electrolytically oxidized (PEO) variants. The results were compared with other methods to detect magnesium degradation (pH, potentiodynamic polarization (PDP), cytocompatibility, SEM (scanning electron microscopy)). The non-ceramized specimens showed the highest degradation rates and vast standard deviations. In contrast, the two PEO samples demonstrated reduced degradation rates with diminished standard deviation. The pH values showed above-average constant levels between 7.4-7.7, likely due to the constant exchange of the fluids. SEM revealed severe cracks on the surface of WE43 after degradation, whereas the ceramized surfaces showed significantly decreased signs of corrosion. PDP results confirmed the improved corrosion resistance of both PEO samples. While WE43 showed slight toxicity in vitro, satisfactory cytocompatibility was achieved for the PEO test samples. In summary, the dynamic test bench constructed in this study enables reliable and simple measurement of Mg degradation to simulate the in vivo environment. Furthermore, PEO treatment of magnesium is a promising method to adjust magnesium degradation.

Development of biomimetic in vitro fatigue assessment for UHMWPE implant materials.

An important research goal in the field of biomaterials lies in the progressive amendment of in vivo tests with suitable in vitro experiments. Such approaches are gaining more significance nowadays because of an increasing demand on life sciences and the ethical issues bound to the sacrifice of animals for the sake of scientific research. Another advantage of transferring the experiments to the in vitro field is the possibility of accurately control the boundary conditions and experimental parameters in order to reduce the need of validation tests involving animals. With the aim to reduce the amount of needed in vivo studies for this cause, a short-time in vitro test procedure using instrumented load increase tests with superimposed environmental loading has been developed at TUD to assess the mechanical long-term durability of ultra-high molecular weight polyethylene (UHMWPE) under fatigue loading in a biological environment.

PEO-generated Surfaces Support Attachment and Growth of Cells In Vitro with No Additional Benefit for Micro-roughness in Sa (0.2-4 µm).

BACKGROUND/AIM: Plasma electrolytic oxidation (PEO), also known as micro-arc oxidation, is a promising electrochemical surface treatment technique for metals which has been used for the generation of various material surfaces and has been the focus of recent biomaterial research. It has been hypothesized that rough PEO surfaces should generally have properties that support cellular attachment and proliferation. However, this has not yet been demonstrated in systematically conducted studies. The present study investigated fibroblast cell proliferation and attachment to ground, electric discharge machining (EDM) and PEO-treated titanium surfaces differing in roughness and porosity. MATERIALS AND METHODS: Three surface variants with 'smoother', 'medium-coarse' and 'rough' surface topographies were generated by PEO and EDM on specimens of titanium alloy (with 6 wt% aluminum and 4 wt% vanadium) for comparison with more smoothly ground specimens. The in vitro effects on cellular attachment and proliferation were determined in 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT), 5-bromo-2'-deoxyuridine (BrdU) and live/dead staining assays with L929 fibroblasts cultivated directly on the metal specimens. Cytocompatibility was determined in accordance with DIN 10993-5/-12 regulations by extract assays. RESULTS: Besides cytocompatibility, all PEO specimens exhibited similar biocompatibility and attachment properties, with vital, spindle-shaped adherent cells growing on the surface, regardless of their surface topology. There were no significant differences in cellular proliferation between the different surfaces and negative controls (cells growing in cell-culture plates). DISCUSSION/CONCLUSION: With no differences in cellular proliferation and attachment between PEO surfaces with different roughness, we find no evidence to support the notion that rougher PEO surfaces are more favorable for cellular growth of fibroblasts in vitro.

Optimized in vitro procedure for assessing the cytocompatibility of magnesium-based biomaterials.

Magnesium (Mg) is a promising biomaterial for degradable implant applications that has been extensively studied in vitro and in vivo in recent years. In this study, we developed a procedure that allows an optimized and uniform in vitro assessment of the cytocompatibility of Mg-based materials while respecting the standard protocol DIN EN ISO 10993-5:2009. The mouse fibroblast line L-929 was chosen as the preferred assay cell line and MEM supplemented with 10% FCS, penicillin/streptomycin and 4mM l-glutamine as the favored assay medium. The procedure consists of (1) an indirect assessment of effects of soluble Mg corrosion products in material extracts and (2) a direct assessment of the surface compatibility in terms of cell attachment and cytotoxicity originating from active corrosion processes. The indirect assessment allows the quantification of cell-proliferation (BrdU-assay), viability (XTT-assay) as well as cytotoxicity (LDH-assay) of the mouse fibroblasts incubated with material extracts. Direct assessment visualizes cells attached to the test materials by means of live-dead staining. The colorimetric assays and the visual evaluation complement each other and the combination of both provides an optimized and simple procedure for assessing the cytocompatibility of Mg-based biomaterials in vitro.