In vitro methods for the evaluation of antimicrobial surface designs.

Bacterial adhesion and subsequent biofilm formation on biomedical implants and devices are a major cause of their failure. As systemic antibiotic treatment is often ineffective, there is an urgent need for antimicrobial biomaterials and coatings. The term "antimicrobial" can encompass different mechanisms of action (here termed "antimicrobial surface designs"), such as antimicrobial-releasing, contact-killing or non-adhesivity. Biomaterials equipped with antimicrobial surface designs based on different mechanisms of action require different in vitro evaluation methods. Available industrial standard evaluation tests do not address the specific mechanisms of different antimicrobial surface designs and have therefore been modified over the past years, adding to the myriad of methods available in the literature to evaluate antimicrobial surface designs. The aim of this review is to categorize fourteen presently available methods including industrial standard tests for the in vitro evaluation of antimicrobial surface designs according to their suitability with respect to their antimicrobial mechanism of action. There is no single method or industrial test that allows to distinguish antimicrobial designs according to all three mechanisms identified here. However, critical consideration of each method clearly relates the different methods to a specific mechanism of antimicrobial action. It is anticipated that use of the provided table with the fourteen methods will avoid the use of wrong methods for evaluating new antimicrobial designs and therewith facilitate translation of novel antimicrobial biomaterials and coatings to clinical use. The need for more and better updated industrial standard tests is emphasized. STATEMENT OF SIGNIFICANCE: European COST-action TD1305, IPROMEDAI aims to provide better understanding of mechanisms of antimicrobial surface designs of biomaterial implants and devices. Current industrial evaluation standard tests do not sufficiently account for different, advanced antimicrobial surface designs, yet are urgently needed to obtain convincing in vitro data for approval of animal experiments and clinical trials. This review aims to provide an innovative and clear guide to choose appropriate evaluation methods for three distinctly different mechanisms of antimicrobial design: (1) antimicrobial-releasing, (2) contact-killing and (3) non-adhesivity. Use of antimicrobial evaluation methods and definition of industrial standard tests, tailored toward the antimicrobial mechanism of the design, as identified here, fulfill a missing link in the translation of novel antimicrobial surface designs to clinical use.

Fabrication of biopolymer-based staple electrospun fibres for nanocomposite applications by particle-assisted low temperature ultrasonication.

We demonstrate the fabrication of staple polymer-based fibres by the ultrasound-assisted processing of electrospun meshes. Bioabsorbable Poly-L-Lactic Acid (PLLA) was electrospun from organic solvent mixtures, yielding continuous fibres with diameters in the range of 244±78 nm. Subsequently, the obtained fibres were sonicated at low temperatures in the presence of nanoparticles in order to obtain fibres with small aspect ratios. The influence of the dispersion medium, the sonication process parameters (temperature and time) and the dimensions of the particles used on the respective length distribution of the obtained nanofibres was investigated. Hexane was identified as an optimal dispersion medium for the system studied in this work. When a cooling bath temperature of 0°C was used, a slight increase in the obtained fibres' average length and distribution was observed as compared to cooling at -80°C (54±43 µm vs 44±31 µm). Moreover, in the presence of hydroxyapatite and hydrophilic and hydrophobic TiO2 nanoparticles in the dispersion medium longer fibres were obtained (44±31 µm, 63±47 µm, and 51±52 µm). Finally, the application of the obtained PLLA-fibre-hydroxyapatite (HA) nanoparticle precursors for the fabrication of a fibre-reinforced Brushite-based cement with high compressive strength is shown. This method of obtaining nanoscaled fibre-reinforced materials opens up a wide range of perspectives for the fabrication of composites for tissue engineering applications.

Evaluation of biocompatibility using in vitro methods: interpretation and limitations.

The in vitro biocompatibility of novel materials has to be proven before a material can be used as component of a medical device. This must be done in cell culture tests according to internationally recognized standard protocols. Subsequently, preclinical and clinical tests must be performed to verify the safety of the new material and device. The present chapter focuses on the first step, the in vitro testing according to ISO 10993-5, and critically discusses its limited significance. Alternative strategies and a brief overview of activities to improve the current in vitro tests are presented in the concluding section.