Biological evaluation of selective laser melted magnesium alloy powder.

PURPOSE: The current study examined magnesium alloy AZ31B specimens manufactured with Additive Manufacturing method (selective laser melting - SLM) to investigate the applicability of this technology for the production of medical devices. METHODS: Osteoblast cells and bacterial biofilm growth ability on specimen was examined and the effect of surface state on corrosion resistance was evaluated by electrochemical and immersion methods. RESULTS: High survival of hFOB cells, as well as a strong tendency for Pseudomonas aeruginosa and Staphylococcus aureus biofilm proliferation on the surface of the tested specimens were shown. SLM-processed AZ31B alloy has a higher corrosion resistance in 0.9% NaCl solution and in a multi-electrolyte saline solution than the material in a conventional form of a rolled sheet. CONCLUSIONS: It has been demonstrated that the strong development of the surface of as-built processed specimens results in a significantly increased corrosion rate, which hinders the production of complex structures in tissue engineering products that support cell ingrowth.

Effect of Scanning and Support Strategies on Relative Density of SLM-ed H13 Steel in Relation to Specimen Size.

Standard experimental research works are aimed at optimization of Selective Laser Melting (SLM) parameters in order to produce material with relative density over 99% and possibly the highest scanning speed. Typically, cuboidal specimens with arbitrarily selected dimensions are built. An optimum set of parameters, determined on such specimens, is used for building parts with variable cross-section areas. However, it gives no guarantee that the density of variable-section parts produced with so selected parameters will be as high as that of the specimens measured during the parameters optimization process. The goal of this work was to improve the process of SLM parameter selection according to the criterion of maximum relative density, based on the example of AISI H13 tool steel (1.2344). A selection method of scanning strategy ensuring relative density of parts over 99%, irrespective of their dimensions, was determined. The specimens were produced using several variants of support structures. It was found that proper selection of the support strategy prevents development of columnar pores.

Application of Ti6Al7Nb Alloy for the Manufacture of Biomechanical Functional Structures (BFS) for Custom-Made Bone Implants.

Unlike conventional manufacturing techniques, additive manufacturing (AM) can form objects of complex shape and geometry in an almost unrestricted manner. AM’s advantages include higher control of local process parameters and a possibility to use two or more various materials during manufacture. In this work, we applied one of AM technologies, selective laser melting, using Ti6Al7Nb alloy to produce biomedical functional structures (BFS) in the form of bone implants. Five types of BFS structures (A1, A2, A3, B, C) were manufactured for the research. The aim of this study was to investigate such technological aspects as architecture, manufacturing methods, process parameters, surface modification, and to compare them with such functional properties such as accuracy, mechanical, and biological in manufactured implants. Initial in vitro studies were performed using osteoblast cell line hFOB 1.19 (ATCC CRL-11372) (American Type Culture Collection). The results of the presented study confirm high applicative potential of AM to produce bone implants of high accuracy and geometric complexity, displaying desired mechanical properties. The experimental tests, as well as geometrical accuracy analysis, showed that the square shaped (A3) BFS structures were characterized by the lowest deviation range and smallestanisotropy of mechanical properties. Moreover, cell culture experiments performed in this study proved that the designed and obtained implant’s internal porosity (A3) enhances the growth of bone cells (osteoblasts) and can obtain predesigned biomechanical characteristics comparable to those of the bone tissue.

The chemical digestion of Ti6Al7Nb scaffolds produced by Selective Laser Melting reduces significantly ability of Pseudomonas aeruginosa to form biofilm.

In our previous work we reported the impact of hydrofluoric and nitric acid used for chemical polishing of Ti-6Al-7Nb scaffolds on decrease of the number of Staphylococcus aureus biofilm forming cells. Herein, we tested impact of the aforementioned substances on biofilm of Gram-negative microorganism, Pseudomonas aeruginosa, dangerous pathogen responsible for plethora of implant-related infections. The Ti-6Al-7Nb scaffolds were manufactured using Selective Laser Melting method. Scaffolds were subjected to chemical polishing using a mixture of nitric acid and fluoride or left intact (control group). Pseudomonal biofilm was allowed to form on scaffolds for 24 hours and was removed by mechanical vortex shaking. The number of pseudomonal cells was estimated by means of quantitative culture and Scanning Electron Microscopy. The presence of nitric acid and fluoride on scaffold surfaces was assessed by means of IR and rentgen spetorscopy. Quantitative data were analysed using the Mann-Whitney test (P ≤ 0.05). Our results indicate that application of chemical polishing correlates with significant drop of biofilm-forming pseudomonal cells on the manufactured Ti-6Al-7Nb scaffolds ( p = 0.0133, Mann-Whitney test) compared to the number of biofilm-forming cells on non-polished scaffolds. As X-ray photoelectron spectroscopy revealed the presence of fluoride and nitrogen on the surface of scaffold, we speculate that drop of biofilm forming cells may be caused by biofilm-supressing activity of these two elements.

Microbial biofilms are able to destroy hydroxyapatite in the absence of host immunity in vitro.

PURPOSE: It is widely thought that inflammation and osteoclastogenesis result in hydroxyapatite (HA) resorption and sequestrum formation during osseous infections, and microbial biofilm pathogens induce the inflammatory destruction of HA. We hypothesized that biofilms associated with infectious bone disease can directly resorb HA in the absence of host inflammation or osteoclastogenesis. Therefore we developed an in vitro model to test this hypothesis. MATERIALS AND METHODS: Customized HA discs were manufactured as a substrate for growing clinically relevant biofilm pathogens. Single-species biofilms of Streptococcus mutans, Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans and mixed-species biofilms of C albicans plus S mutans were incubated on HA discs for 72 hours to grow mature biofilms. Three different non-biofilm control groups also were established for testing. HA discs were then evaluated by means of scanning electron microscopy, micro-computed tomography metrotomography, x-ray spectroscopy, and confocal microscopy with planimetric analysis. In addition, quantitative cultures and pH assessment were performed. Analysis of variance was used to test for significance between treatment and control groups. RESULTS: All investigated biofilms were able to cause significant (P < .05) and morphologically characteristic alterations in HA structure as compared with controls. The highest number of alterations observed was caused by mixed biofilms of C albicans plus S mutans. S mutans biofilm incubated in medium with additional sucrose content was the most detrimental to HA surfaces among single-species biofilms. CONCLUSIONS: Our findings suggest that direct microbial resorption of bone is possible in addition to immune-mediated destruction, which has important translational implications for the pathogenesis of chronic bone infections and for targeted antimicrobial therapeutics.

The ability of S.aureus to form biofilm on the Ti-6Al-7Nb scaffolds produced by Selective Laser Melting and subjected to the different types of surface modifications.

The Gram-positive coccus, Staphylococcus aureus, is the leading etiologic agent of limb and life-threatening biofilm-related infections in the patients following the orthopaedic implantations. The aim of the present paper is to estimate the ability of S. aureus to form biofilm on titanium alloy (Ti-6Al-7Nb) scaffolds produced by Selective Laser Melting (SLM) and subjected to the different types of surface modifications, including ultrasonic cleaning and chemical polishing. The results obtained indicate significantly the decreased ability of S.aureus to form biofilm on the surface of scaffolds subjected to the chemical polishing in comparison to the scaffolds cleaned ultrasonically. The data provided can be useful for future applications of the SLM technology in production of Ti-6Al-7Nb medical implants.