Pseudomonas aeruginosa biofilm formation and slime excretion on antibiotic-loaded bone cement

Background Infection is an infrequent but serious complication of prosthetic joint surgery. These infections will usually not clear until the implant is removed and re-implantation has a high failure rate, especially when Pseudomonas aeruginosa is involved. Material and methods We examined Pseudomonas aeruginosa biofilm formation on plain and gentamicin-loaded bone cement with confocal scanning laser microscopy (CSLM). Two different stains were applied in order to visualize and quantify the distribution of bacterial cells and extracellular polymeric substances (slime) from the bone cement surface to the top of the biofilm. Staining with LIVE/DEAD viability stain differentiated between live and dead bacteria within the biofilm, and slime production was evaluated after staining with Calcofluor white. Results CSLM showed that the biofilm was a nonuniform structure of variable thickness, with differences in local bacterial cell and slime densities. Incorporation of gentamicin in bone cement resulted in a 44% reduction in bacterial viability, while the slime density increased significantly. In addition, conventional plate counting showed the development of small-colony variants on gentamicin-loaded bone cement with a decreased sensitivity for gentamicin (MIC: 8 mg/L), as compared with normal-sized colonies taken from plain and gentamicin-loaded bone cement (MIC: 3 mg/L). The enhanced slime production on antibiotic-loaded bone cement, together with the formation of small-colony variants, resulted in decreased susceptibility to antibiotics-probably concomitant with the onset of persistent and relapsing infections. Interpretation In the clinical situation, our findings help to explain the frequent re-implantation failure of joint replacements infected with P. aeruginosa when the procedure has been performed using antibiotic-loaded bone cement

Translational and rotational dynamics of a large buoyant sphere in turbulence

We report experimental measurements of the translational and rotational dynamics of a large buoyant sphere in isotropic turbulence. We introduce an efficient method to simultaneously determine the position and (absolute) orientation of a spherical body from visual observation. The method employs a minimization algorithm to obtain the orientation from the 2D projection of a specific pattern drawn onto the surface of the sphere. This has the advantages that it does not require a database of reference images, is easily scalable using parallel processing, and enables accurate absolute orientation reference. Analysis of the sphere’s translational dynamics reveals clear differences between the streamwise and transverse directions. The translational autocorrelations and PDFs provide evidence for periodicity in the particle’s dynamics even under turbulent conditions. The angular autocorrelations show weak periodicity. The angular accelerations exhibit wide tails, however without a directional dependence

The role of small-colony variants in failure to diagnose and treat biofilm infections in orthopedics

Biomaterial-related infection of joint replacements is the second most common cause of implant failure, with serious consequences. Chronically infected replacements cannot be treated without removal of the implant, as the bio film mode of growth protects the bacteria against antibiotics. This review discusses bio film formation on joint replacements and the important clinical phenomenon of small-colony variants (SCVs). These slow-growing phenotypic variants often remain undetected or are misdiagnosed using hospital microbiological analyses due to their unusual morphological appearance and biochemical reactions. In addition, SCVs make the infection difficult to eradicate. They often lead to recurrence since they respond poorly to standard antibiotic treatment and can sometimes survive intracellularly

Concepts for increasing gentamicin release from handmade bone cement beads

Background and purpose Commercial gentamicin-loaded bone cement beads (Septopal) constitute an effective delivery system for local antibiotic therapy. These beads are not available in all parts of the world, and are too expensive for frequent use in others. Thus, orthopedic surgeons worldwide make antibiotic-loaded beads themselves. However, these beads are usually not as effective as the commercial beads because of inadequate release kinetics. Our purpose was to develop a simple, cheap, and effective formulation to prepare gentamicin-loaded beads with release properties and antibacterial efficacy similar to the commercially ones. Methods Acrylic beads were prepared with variable monomer content: 100% (500 μL/g polymer), 75%, and 50% to increase gentamicin release through creation of a less dense polymer matrix. Using the optimal monomer content, different gel-forming polymeric fillers were added to enhance the permeation of fluids into the beads. Polyvinylpyrrolidone (PVP) 17 was selected as a suitable filler; its concentration was varied and the antibiotic release and antibacterial efficacy of these beads were compared with the corresponding properties of the commercial ones. Results Gentamicin release rate and the extent of release from beads prepared with 50% monomer increased when the PVP17 content was increased. Beads with 15 w/w% PVP17 released 87% of their antibiotic content. This is substantially more than the gentamicin release from Septopal beads (59%). Acrylic beads with 15 w/w% PVP17 reduced bacterial growth by up to 93%, which is similar to the antibacterial properties of the commercial ones. Interpretation A simple, cheap, and effective formulation and preparation process has been described for hand-made gentamicin-releasing acrylic beads, with better release kinetics and with antibacterial efficacy similar to that of the commercial ones

De verbintenisrechtelijke bescherming van de kleine opdrachtnemer

Dit boek gaat over het verbintenisrechtelijke beschermingsniveau van de kleine opdrachtnemer ten aanzien van de thema’s loon, aansprakelijkheid en opzegging. Op de verhouding tussen deze zelfstandig ondernemer en een niet-particuliere opdrachtgever zijn in principe geen beschermende rechtsregels van toepassing. Dit uitgangspunt wordt ten aanzien van een aantal opdrachtnemers als een steeds urgenter maatschappelijk probleem gezien, terwijl er ook een heel arsenaal is aan opdrachtnemers die niet of minder in een kwetsbare positie (lijken te) verkeren. Hierdoor is het vinden van regels voor opdrachtnemers ten aanzien van zowel het huidige als toekomstige recht uiterst complex. In dit kader analyseert de auteur per thema: de regeling inzake de opdracht, de bijzondere rechtsregels die van toepassing zijn verklaard op de overeenkomst van opdracht, de (open) normen die uit het algemene verbintenissenrecht voortvloeien en (ter inspiratie) de bepalingen die gelden voor de werknemer, aannemer, handelsagent en huurder. De resultaten van dit onderzoek bieden niet alleen een inkijk in het verbintenisrechtelijke beschermingsniveau van de kleine opdrachtnemer, maar maken ook een gefundeerde afweging mogelijk ten aanzien van de vraag of dit beschermingsniveau wenselijk en gerechtvaardigd is