Effect of S53P4 bone substitute on staphylococcal adhesion and biofilm formation on other implant materials in normal and hypoxic conditions.

To study the effect of bioactive glass bone substitute granules (S53P4) on bacterial adhesion and biofilm formation on other simultaneously used implant materials and the role of the hypoxic conditions to the adhesion. Bacterial and biofilm formation were studied on materials used both in middle ear prostheses and in fracture fixtures (titanium, polytetrafluoroethylene, polydimethylsiloxane and bioactive glass plates) in the presence or absence of S53P4 granules. The experiments were done either in normal atmosphere or in hypoxia simulating atmospheric conditions of middle ear, mastoid cavity and sinuses. We used two collection strains of Staphylococcus aureus and Staphylococcus epidermidis. In the presence of bioglass and hypoxic conditions the adhesion of the planktonic bacterial cells was decreased for most of the materials. The biofilm formation was decreased for S. epidermidis on titanium and polydimethylsiloxane in both atmospheric conditions and on bioglass plates in normoxia. For S. aureus the biofilm formation was decreased on bioglass plates and polytetrafluoroethylene in normoxia. Hypoxia produces a decrease in the biofilm formation only for S. aureus on polytetrafluoroethylene and for S. epidermidis on bioglass plates. However, in none of the cases bioactive glass increased the bacterial or biofilm adhesion. The presence of bioglass in normoxic and hypoxic conditions prevents the bacterial and biofilm adhesion on surfaces of several typical prosthesis materials in vitro. This may lead to diminishing postoperative infections, however, further in vivo studies are needed.

Effects of S53P4 bioactive glass on osteoblastic cell and biomaterial surface interaction.

To study the effect of bioactive glass bone substitute granules (S53P4) and hypoxic atmospheric conditions on human osteoblastic cell adhesion on different biomaterials. Cellular adhesion and cytoskeletal organization were studied on titanium, polytetrafluoroethylene, polydimethylsiloxane and S53P4 plates in the presence or absence of S53P4 granules. Cells used were human osteoblast-like SaOS-2 cells. The experiments were done either in normal atmospheric conditions or in hypoxia which simulates conditions prevailing in chronically infected bone or bone cavities. Vinculin-containing focal adhesions, organization of actin cytoskeleton and nuclear staining of cells on biomaterial surfaces were studied at 4.5 h, 2 and 4 days. In normoxic conditions S53P4 granules alkalinized the cell culture medium but cellular adhesion and cytoskeletal organization were usually not affected by their presence. Hypoxic conditions associated with lower pH and impaired cellular adhesion, vinculin-containing focal adhesion formation and rearrangement of the actin filaments to actin cytoskeleton. On most materials studied in hypoxic conditions, however, S53P4 granules prevented this impairment of cellular adhesion and cytoskeletal reorganization. The S53P4 granules promote the adhesion of SaOS-2 cells to various biomaterial surfaces especially in hypoxic conditions, in which S53P4 granules increase pH. The presence of S53P4 granules may protect biomaterial surface from bacterial colonization and promote osteointegration of implants used together with S53P4 granules for fixation and weight bearing.

Effect of surface roughness and sterilization on bacterial adherence to ultra-high molecular weight polyethylene.

Sterilization with ethylene oxide (EO) and gas plasma (GP) are well-known methods applied to ultra-high molecular weight polyethylene (UHMWPE) surfaces in the belief that they prevent major material changes caused by gamma irradiation. However, the influence of these surface sterilization methods on bacterial adherence to UHMWPE is unknown. UHMWPE samples with various degrees of roughness (0.3, 0.8 and 2.0 µm) were sterilized with either GP or EO. The variations in hydrophobicity, surface free energy and surface functional groups were investigated before and after sterilization. Sterilized samples were incubated with either Staphylococcus aureus or Staphylococcus epidermidis in order to study bacterial adherence to these materials. Fewer bacteria adhered to UHMWPE after sterilization with EO than after sterilization with GP, especially to the smoothest surfaces. No changes in chemical composition of the UHMWPE surface due to sterilization were observed using X-ray photoemission spectroscopy analysis. The decreased bacterial adherence to UHMWPE found at the smoothest surfaces after sterilization with EO was not directly related to changes in chemical composition. Increased bacterial adherence to rougher surfaces was associated with increased polar surface energy of EO-sterilized surfaces.

Detection of lfrA and tap efflux pump genes among clinical isolates of non-pigmented rapidly growing mycobacteria.

This study was performed to detect LfrA and Tap efflux pumps among clinical isolates of non-pigmented rapidly growing mycobacteria (NPRGM). Gene detection was performed using polymerase chain reaction (PCR) with specific primers designed for each gene. Susceptibility of the strains to doxycycline, tigecycline and ciprofloxacin was analysed using the broth microdilution reference technique. In total, 166 clinical isolates were included in the study. The lfrA gene was detected in four strains (2.4%), comprising two strains of Mycobacterium chelonae (6.7% of this species), one Mycobacterium fortuitum (1.1%) and one Mycobacterium mucogenicum (14.3%). The tap gene was detected in 109 strains (65.7%), comprising 3 Mycobacterium abscessus (33.3%), 12 M. chelonae (40%), 75 M. fortuitum (84.3%), 2 Mycobacterium mageritense (40%), 15 Mycobacterium peregrinum (68.2%), 1 Mycobacterium alvei and 1 Mycobacterium porcinum; no strains of M. mucogenicum were tap-positive. No differences between tap-positive and -negative strains were observed for resistance to doxycycline (Fisher's exact test, P=0.055). lfrA is rare among clinical isolates of NPRGM, whilst tap is found more commonly. No correlation was detected between the presence of the efflux pumps and resistance to quinolones or tetracyclines.

In-vitro evaluation of the adhesion to polypropylene sutures of non-pigmented, rapidly growing mycobacteria.

The ability of non-pigmented, rapidly growing mycobacteria (NPRGM) to attach to polypropylene sutures was evaluated using an in-vitro assay. Thirty clinical isolates and five culture collection strains of NPRGM, together with Staphylococcus epidermidis ATCC 35983, were tested. Mycobacterium fortuitum and Mycobacterium chelonae showed the highest attachment ability, which differed significantly from the results obtained with Mycobacterium peregrinum. According to these results, NPRGM are able to attach to polypropylene sutures, and the species implicated most frequently in human infection showed increased levels of attachment in comparison with the other mycobacteria studied.

New method for coating tympanostomy tubes to prevent tube occlusions.

OBJECTIVE: tympanostomy tube insertion is currently the most common surgical procedure requiring general anesthesia performed on children. Occlusion of the tube and prolonged otorrhea through the tube are typical problems associated with the use of middle-ear ventilation tubes. In this study, a new method for coating ventilation tubes is introduced that prevents occlusion of the tube lumen by granulation tissue, blood clot or pus. METHODS: human serum albumin (HSA) was used to coat standard tympanostomy tubes of different materials. Fibronectin, a typical protein in serum and exudates and one of the most adhesive glycoproteins, was used as a model representative of exudates of the ear. RESULTS: when compared with the binding on uncoated tubes, the binding of fibronectin on HSA-coated tubes was inhibited from 59 to 85%, depending on the tube material used. CONCLUSIONS: HSA-coating markedly reduced the binding of fibronectin on tube surfaces in vitro. The study shows the potential role of HSA-coating in preventing the adherence of foreign material to tympanostomy tubes and reducing tube occlusions.