TiO2 nanotube composite layers as delivery system for ZnO and Ag nanoparticles - an unexpected overdose effect decreasing their antibacterial efficacy.

Enhancement of biocompatibility and antibacterial properties of implant materials is potentially beneficial for their practical value. Therefore, the use of metallic and metallic oxide nanoparticles as antimicrobial coatings components which induce minimized antibacterial resistance receives currently particular attention. In this work, TiO2 nanotubes layers loaded with ZnO and Ag nanoparticles were designed for biomedical coatings and delivery systems and evaluated for antimicrobial activity. TiO2 nanotubes themselves exhibited considerable and diameter-dependent antibacterial activity against planktonic Staphylococcus epidermidis cells but favored bacterial adhesion. Loading of nanotubes with moderate amount of ZnO nanoparticles significantly diminished S. epidermidis cell adhesion and viability just after 1.5h contact with modified surfaces. However, an increase of loaded ZnO amount unexpectedly altered the structure of nanoparticle-nanolayer, caused partial closure of nanotube interior and significantly reduced ZnO solubility and antibacterial efficacy. Co-deposition of Ag nanoparticles enhanced the antibacterial properties of synthesized coatings. However, the increase of ZnO quantity on Ag nanoparticles co-deposited surfaces favored the adhesion of bacterial cells. Thus, ZnO/Ag/TiO2 nanotube composite layers may be promising delivery systems for combating post-operative infections in hard tissue replacement procedures. However, the amount of loaded antibacterial agents must be carefully balanced to avoid the overdose and reduced efficacy.

Vascular prostheses with covalently bound gentamicin and amikacin reveal superior antibacterial properties than silver-impregnated ones--an in vitro study.

OBJECTIVE: This study aims to compare the antibacterial activities of vascular prostheses: silver-impregnated and modified with covalently immobilised antibiotics. MATERIALS AND METHODS: Six types of protein-sealed vascular prostheses were modified with amikacin and gentamicin according to the method described in the Polish Patent Office. Their antimicrobial properties were estimated against 14 reference and clinical strains and compared with those of InterGard Silver grafts. Cytotoxicity of the tested grafts was estimated against human skin fibroblasts. RESULTS: Prostheses modified with antibiotics in a stable covalent mode were found to be much more effective against bacterial growth and biofilm formation, as well as in case of methicillin-resistant Staphylococcus aureus (MRSA), than InterGard Silver. They inhibited the bacterial growth for at least 30 days, without losing higher than 10% of the initial amount of its drug content. They were also good, non-toxic matrices for growth of human skin fibroblasts. CONCLUSIONS: Prostheses modified with covalently immobilised antibiotic according to our technique are much more effective than InterGard Silver at protection against bacterial growth. They are also compatible with human skin fibroblasts.

Microbial conversion of daunomycin wastes in unsteril soil inoculated with Bjerkandera adusta R59.

Effect of cooperation between native soil microorganisms and white-rot fungus Bjerkandera adusta R59 on degradation of daunomycin post-production wastes in soil was described. Pure cultures of B. adusta R59 strain were capable to decolorize and decompose that cytostatic xenobiotic in liquid media. Presence of R59 strain in studied daunomycin waste/soil systems increased the rate of the antibiotic conversion. The markers of that process were changes of waste biomass or daunomycin concentration (in pulp) and phenolics level and peroxidase activity (in effluent). It was shown that daunomycin in the wastes may be metabolized even up to 20% of its initial concentration. This effect was conjugated with the propagation of native soil microorganisms (microfungi and bacteria) more significant than in parallel system without R59 strain.

Antibacterial activity of gentamicin-bonded gelatin-sealed polyethylene terephthalate vascular prostheses.

OBJECTIVES: To create an antibiotic-modified vascular prosthesis with a prolonged bactericidal activity, susceptible to endothelialisation. METHODS: We used a covalent method of gentamicin sulphate immobilisation to polyethylene terephthalate prosthesis sealed with gelatin. Antibacterial activity was assayed in Luria-Bertani medium against Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa strains. Prosthesis endothelialisation was performed using bovine aorta endothelial cells (BAEC). RESULTS: Gentamicin was bound to vascular prostheses in the amount of 12g per kg of prosthesis. Ninety-seven percent of antibiotic bound in covalent way and remained on the biomaterial for at least 30 days during shaking in PBS solution. Gentamicin-modified prostheses exerted bactericidal or bacteriostatic effect on growth of clinical and reference bacterial strains, prevented biofilm formation and were highly susceptible to endothelialisation. BAEC viability exceeded 90%, which indicated that gentamicin-vascular prostheses were not toxic for these cells. CONCLUSIONS: Covalent gentamicin immobilisation resulted in effective antibacterial protection of vascular prostheses against clinical and reference strains of S. aureus, E. coli and P. aeruginosa and allowed for a strong adherence of endothelial cells to antibiotic-modified prostheses.

Extracellular enzyme activities of Bjerkandera adusta R59 soil strain, capable of daunomycin and humic acids degradation.

Geotrichum-like strain R59, the anamorphic form of the white-rot fungus, Bjerkandera adusta, was isolated from soil. It was found to completely decolorize and degrade 10% daunomycin post-production effluent during 10 days of incubation at 26 degrees C. Strain R59 produced only low levels of ligninolytic enzymes when grown on wheat straw- or beech sawdust-containing media, but in the presence of humic acids derived from brown coal it synthesized significant amounts of laccase and lipase. This phenomenon was coupled with the fungus entering the idiophase and the appearance of aerial mycelium. B. adusta strain R59 was found to completely decolorize 0.03% humic acids from brown coal and lessive soil and to partially decolorize humic acids isolated from a chernozem during 14 days of growth. This ability of strain R59 could be useful in constructing a new generation of biologically active filters for the purification of humic acids-contaminated drinking waters.

The novel non-glycosylated invertase from Candida utilis (the properties and the conditions of production and purification).

The Candida utilis yeast, which is cultivated in liquid media enriched with saccharose, synthesizes the well-known invertase of 300 kDa (EC 3.2.1.26). This enzyme is present both intracellularly in the periplasmic space and extracellularly in the culture broth. However, it was determined that the same C. utilis strain cultured in certain conditions is simultaneously capable of producing another, still unknown form of invertase with a molecular mass of 60 kDa. The presence of the latter enzymatic form was detected in cells as well as in the liquid culture medium. Both invertase forms were purified using a three-step process (ion-exchange chromatography, affinity chromatography, and preparative column electrophoresis) and named, due to their different migration ratio in polyacrylamide gel electrophoresis, F-form (Fast; 60 kDa) and S-form (Slow; 300 kDa). The F-form of invertase was found to be nonglycosylated as opposed to the well-known S-form of invertase from the same source. The physicochemical properties of the F-form of invertase (isoelectric point, substrate specificity, pH, and temperature optima) were determined and compared with those of the S-form of the enzyme.