Abstract
Bacterial infection of biomaterials is a major concern in medicine, and different kinds of antimicrobial biomaterial have been developed to deal with this problem. To test the antimicrobial performance of these biomaterials, the airborne bacterial assay is used, which involves the formation of biohazardous bacterial aerosols. We here describe a new experimental set-up which allows safe handling of such pathogenic aerosols, and standardizes critical parameters of this otherwise intractable and strongly user-dependent assay. With this new method, reproducible, thorough antimicrobial data (number of colony forming units and live-dead-stain) was obtained. Poly(oxonorbornene)-based Synthetic Mimics of Antimicrobial Peptides (SMAMPs) were used as antimicrobial test samples. The assay was able to differentiate even between subtle sample differences, such as different sample thicknesses. With this new set-up, the airborne bacterial assay was thus established as a useful, reliable, and realistic experimental method to simulate the contamination of biomaterials with bacteria, for example in an intraoperative setting.
Highlights
Antimicrobial in-vitro testing is crucial for the design, development and in-vivo performance prediction of biomaterials
It turns out that these apparently trivial modifications are crucial for the data quality. We have evaluated this new set-up with antimicrobial polymer coatings, with poly(oxonorbornene)-based synthetic mimics of antimicrobial peptides (SMAMPs) [8,9,10,11,12,13]
The following requirements had to be fulfilled: N obtaining a homogenous bacterial suspension with a welldefined optical density and number of colony forming units; N reproducible spraying of a constant amount of the bacterial suspension; N multiple specimens of the material should be sprayed in each experiment; N the bacterial aerosols formed must not enter the laboratory; N after the experiment, the entire contaminated set-up should be autoclavable without damage to any of its components
Summary
Antimicrobial in-vitro testing is crucial for the design, development and in-vivo performance prediction of biomaterials. To get meaningful data, it is important to perform these antimicrobial assays with clinically relevant bacterial strains, and to simulate realistic infection scenarios. The sources of contamination found within operating theaters are diverse. They comprise, among others, microbial transport through moving medical personnel, locally formed aerosols (including sneezing/ coughing), or building-related sources such as ventilation and air condition [1,2,3,4]. All in all, ‘Sterile implant surgery may be considered a myth’ [2]. In this context, the airborne infection pathway is of particular relevance, and there is a need for an airborne antimicrobial assay that simulates non-contact microbial infection
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