The antimicrobial effect of calcium-doped titanium is activated by fibrinogen adsorption.

Directly targeting bacterial cells is the present paradigm for designing antimicrobial biomaterial surfaces and minimizing device-associated infections (DAIs); however, such pathways may create problems in tissue integration because materials that are toxic to bacteria can also be harmful to mammalian cells. Herein, we report an unexpected antimicrobial effect of calcium-doped titanium, which itself has no apparent killing effect on the growth of pathogenic bacteria (Pseudomonas aeruginosa, Pa, ATCC 27853) while presenting strong inhibition efficiency on bacterial colonization after fibrinogen adsorption onto the material. Fine X-ray photoelectron spectroscopy and Fourier-transform infrared spectroscopy analyses reported calcium-dependent shifts of the binding energy in nitrogen and oxygen involved groups and wavenumbers in the amide I and II bands of the adsorbent fibrinogen, demonstrating that locally delivered calcium can react with the carboxy-terminal regions of the Aα chains and influence their interaction with the N-termini of the Bß chains in fibrinogen. These reactions facilitate the exposure of the antimicrobial motifs of the protein, indicating the reason for the surprising antimicrobial efficacy of calcium-doped titanium. Since protein adsorption is an immediate intrinsic step during the implantation surgery, this finding may shift the present paradigm on the design of implantable antibacterial biomaterial surfaces.

Critical parameters in cultivation of experimental biofilms using the example of Pseudomonas fluorescens.

Formation and treatment of biofilms present a great challenge for health care and industry. About 80% of human infections are associated with biofilms including biomaterial centered infections, like infections of prosthetic heart valves, central venous catheters, or urinary catheters. Additionally, biofilms can cause food and drinking water contamination. Biofilm research focusses on application of experimental biofilm models to study initial adherence processes, to optimize physico-chemical properties of medical materials for reducing interactions between materials and bacteria, and to investigate biofilm treatment under controlled conditions. Exploring new antimicrobial strategies plays a key role in a variety of scientific disciplines, like medical material research, anti-infectious research, plant engineering, or wastewater treatment. Although a variety of biofilm models exist, there is a lack of standardization for experimental protocols, and designing experimental setups remains a challenge. In this study, a number of experimental parameters critical for material research have been tested that influence formation and stability of an experimental biofilm using the non-pathogenic model strain of Pseudomonas fluorescens. These parameters include experimental time frame, nutrient supply, inoculum concentration, static and dynamic cultivation conditions, material properties, and sample treatment during staining for visualization of the biofilm. It was shown, that all tested parameters critically influence the experimental biofilm formation process. The results obtained in this study shall support material researchers in designing experimental biofilm setups.

How Nanotopography-Induced Conformational Changes of Fibrinogen Affect Platelet Adhesion and Activation.

The conformational state of adsorbed human plasma fibrinogen (HPF) has been recognized as the determinant factor in platelet adhesion and thrombus formation on blood-contacting biomaterials. Studies have highlighted the ability to control the HPF conformation merely by tailoring surface nanotopographical features. However, a clear relationship between the conformational changes of adsorbed HPF and the degree of platelet adhesion and activation achieved with different surface nanotopographies is still unclear. Here, we examined HPF assembly characteristics on nanostructured polybutene-1 (PB-1) surfaces with nanosized lamellar crystals (LCs), needle-like crystals (NLCs), and a nanostructured high-density polyethylene (HDPE) surface with shish-kebab crystals (SKCs), at a biologically relevant HPF concentration. By exposing the nanostructured surfaces with preadsorbed HPF to human platelets, significant differences in platelet response on LCs/SKCs and NLCs were identified. The former presented a uniform monolayer in the advanced stage of activation, whereas the latter exhibited minimal adhesion and the early stage of activation. Distinct platelet response was related to the postadsorption conformational changes in HPF, which were confirmed by topography-dependent shifts of the amide I band in attenuated total reflection-Fourier transform infrared (ATR-FTIR) analysis. Supported by atomic force microscopy (AFM) characterization, we propose that the mechanism behind the nanotopography-induced HPF conformation is driven by the interplay between the aspect ratios of polymeric crystals and HPF. From the biomedical perspective, our work reveals that surface structuring in a nanoscale size regime can provide a fine-tuning mechanism to manipulate HPF conformation, which can be exploited for the design of thromboresistant biomaterials surfaces.

Quantifying the relationship between surfaces' nano-contact point density and adhesion force of Candida albicans.

It has been recently recognized that controlled surface structuring on the nanometer scale is a successful strategy to endow different materials with antimicrobial properties. Despite many studies on bacterial interactions with nanostructured surfaces, a quantitative link between surface topography and bacterial adhesion is still missing. To quantitatively link cell adhesion data with topographical surface parameters, we performed single-cell spectroscopy on chemically identical surfaces with controlled nano-contact point density achieved by immobilization of gold nanoparticles (AuNP) on gold thin films. Such materials surfaces have previously shown antimicrobial (anti-adhesive) efficacy towards Gram-negative Escherichia coli cells. In the current study, the influence of nano-structured surfaces on the surface coverage and adhesion forces of clinically relevant Candida albicans (C. albicans), the fungus primarily associated with implant infections, was investigated to validate their antimicrobial potency against different microbial cells. The adhesion forces of C. albicans cells to nanostructured surfaces showed a decreasing trend with decreasing contact-point density and correlated well with the results of the respective C. albicans cell counts. The surfaces with the lowest contact-point density, 25 AuNP/µm², resulted in an average adhesion force of 5 nN, which was up to 5 times lower compared to control and 61 AuNP/µm² surfaces. Further, detailed analyses of force-distance curves revealed that the work of adhesion, and thus the energy required to remove the C. albicans cell from the surface is up to 10 times lower on 25 AuNP/µm² surfaces compared to unstructured surfaces. These findings show that a controlled tuning of nanostructured surfaces in terms of accessible nano-contact points is crucial to generate surface structures with enhanced antimicrobial properties. The gained knowledge can be further exploited for the design of biomaterials surfaces to prevent adhesion of some most commonly encountered pathogens.