Quality assurance for antimicrobial devices.

Quality assurance (QA) of products with antimicrobial claims is still a virgin field for medical devices. As the market pushes the development of new devices with antimicrobial properties there is rapidly growing interest in antimicrobial testing. QA systems for antimicrobial device testing are discussed here.

[Nanoparticulate silver. A new antimicrobial substance for bone cement]. / Nanopartikuläres Silber. Ein neues antimikrobielles Agens für Knochenzement.

BACKGROUND: Multiresistant bacteria have become an important problem in prosthetic joint infections. Their frequent resistance against gentamicin, which is commonly used in antibiotic-loaded bone cements, makes a new prophylaxis necessary. METHODS: PMMA-cement was loaded with 1% nanoparticulate silver and its antibacterial activity tested in vitro against gentamicin-resistant MRSE and MRSA strains as well as being compared to the activity of plain and gentamicin-loaded bone cements. A quantitative elution testing was also done to study the potentially cytotoxic effects of NanoSilver cement. RESULTS: Unloaded and PMMA-cement loaded with 2% gentamicin did not exhibit any antibacterial activity against MRSE and MRSA. At 1%, NanoSilver cement completely inhibited the proliferation of MRSA and MRSE. NanoSilver bone cement did not show any significant differences compared to the non-toxic control group. CONCLUSIONS: If these promising in vitro results can be confirmed in vivo, NanoSilver bone cement may be of considerable value in total joint arthroplasty.

Thrombogenicity testing of central venous catheters in vitro.

To date there have been no standard methods for assessing the thrombogenicity of central venous catheters. A procedure for testing the thrombogenicity of intravenous lines such as the silver-impregnated catheter by continuous blood flow in vitro was therefore developed. For this test, fresh blood was drawn from healthy human donors and anti-coagulated with sodium citrate (1:9). All material tested (catheter tubes with and without silver manufactured in the same way, polyethylene tubes and tubes with potentially thrombogenic material) were perfused through their lumen with anticoagulated blood for up to 31 hours. Blood samples were collected at different times from the test system at sites before and after the perfusion of the test catheters. The hemoglobin concentration, erythrocyte, leukocyte and thrombocyte counts and markers for thrombin activation (thrombin-antithrombin III-complex, F1 + 2)-prothrombin fragments) and for hyperfibrinolysis (d-dimers) were determined. No thrombin activation or signs of hyperfibrinolysis were detected in any material tested. Polyethylene tubes were found to cause hemolysis, as shown by a decrease in hemoglobin content from 15 g% to 4.5 g%. Tecothane tubes with and without silver did not induce hemolysis.

The Erlanger silver catheter: in vitro results for antimicrobial activity.

The antimicrobial activity of a silver-impregnated polymer catheter (the Erlanger silver catheter) was demonstrated by determining the microbial adhesion to the surface of the catheter and by measuring the rate of proliferation (viability) of microorganisms at this site. On the surface of a catheter impregnated with silver, according to previously described methods, the bacterial adhesion of Staphylococcus epidermidis is reduced by 28-40%. Bacterial proliferation on the surface of the catheter and biofilm production are also substantially reduced by the elution of free silver ions from the catheter matrix. Bacteriostatic and bactericidal activities can be determined. The antimicrobial efficacy of the silver catheter is not reduced by blood components. There is no loss in antimicrobial activity for weeks after preincubation in water or phosphate buffered saline. The antimicrobial activity depends on the extent of the active silver surface.

Analysis of the acute cytotoxicity of the Erlanger silver catheter.

The Erlanger silver catheter consists of a new form of polyurethane, which contains finely dispersed metallic silver. The aim of this study was to establish the biocompatibility of this intravenous catheter by investigating the acute cytotoxicity of extracts from the Erlanger silver catheter on human fibroblasts and lymphocytes. Extracts of the Erlanger silver catheter were not cytotoxic for MRC-5 human fibroblasts nor for sensitized phytohemagglutinin (PHA)-stimulated human lymphocytes. The addition of silver powder of up to 2% by weight to the basic catheter polyurethane Tecothane led to no increase in acute cytotoxicity in comparison with untreated Tecothane. The Erlanger silver catheter is a new intravenous catheter with good biocompatibility.

A new scoring system for the clinical diagnosis of catheter-related infections.

It is difficult to make the clinical diagnosis of catheter-related infections using the available and established definitions of the HICPAC (Hospital Infection Control Practices Advisory Committee) of the CDC (Centers for Disease Control, definitions of nosocomial infections). The scoring system shown here is a modification of these definitions and has enabled the causal relationship between the catheter and clinical episodes of systemic infections to be quantitatively graded. The scoring system included the following criteria: height and rate of rise of body temperature, attendant shivering, identification of pathogens in blood and/or catheter tip cultures, improvement in the clinical course after catheter removal, signs of catheter exit site inflammation and results of diagnostic tests for other possible sources of infection. These criteria were graded using points and weighted according to their specificity. The comparative evaluation of 65 episodes of systemic infections using the scoring system and the diagnostic criteria of HICPAC showed agreement in 85%. No case was graded "false-negative." In nine of ten false-positive cases additional findings supported the presence of a catheter-associated infection. This scoring system appears, therefore, to be more sensitive than existing diagnostic criteria, without loss of specificity.

Reduced rates of catheter-associated infection by use of a new silver-impregnated central venous catheter.

A central venous catheter with a new form of silver impregnation of the internal and external surfaces was investigated for antimicrobial activity and tolerance in patients in a controlled comparative, prospective and randomized clinical study. Commercially available catheters with no antimicrobial activity were used as controls. One hundred sixty-five catheters were included in the final evaluation. All catheters were percutaneously inserted for the first time with a duration of > or = 5 days and a microbiological examination of the catheter tip. Catheter location (> 90% internal jugular vein), mean duration of catheterization (8-9 days), patients' age and diagnosis were comparable in both groups. Silver-impregnated catheter tips showed an incidence of colonization in 14.2/1000 catheter days and control catheters in 22.8/1000 catheter days. This represents a reduction of 37.7%. Catheter-associated infections were diagnosed in the silver group in 5.26/1000 catheter days and 18.34/1000 catheter days in the control group, indicating a reduction rate of 71.3% (P < 0.05, chi 2-test). No complications or side effects were documented in either group.

Reducing catheter-associated infections with silver-impregnated catheters in long-term therapy of children.

Central venous long-term catheters offer reliable, large-lumen vascular access with high flow rates for delivery of nutrition or for cell-containing infusions and perfusions. Catheter-associated infections (CAI) pose the greatest threat to such vascular access, despite existing preventive measures. In this article one prospective and one retrospective study of CAI in pediatric therapy are presented. Study I: A retrospective investigation from 1990 through 1995 of 60 conventional long-term catheters in 50 patients. The total number of days in which the catheters were in place was 11,818. The calculated CAI incidence was 1 per 1,000 days of catheter insertion. Bacteriologically demonstrated CAI (identical isolate on the catheter tip and in a blood culture) occurred in three instances (5%). Five cases (8.3%) were diagnosed with a therapy-resistant, septic clinical picture. Study II: A prospective, randomized comparison of long-term silver-impregnated (Erlanger silver catheters) and control catheters (Quinton Instrument Co.) was made with 41 patients (20 with a silver catheter, 21 with a Quinton catheter). To date, the silver catheters have been distinguished by sterile bacteriological findings, whereas three cases of CAI have been demonstrated with the comparative catheters. One patient recently underwent intensive care after becoming unstable with signs of septic shock and demonstrable Pseudomonas aeruginosa, and two other patients manifested coagulase-negative staphylococci on the catheter tips. In three of nine control catheters an incidence of 1.18 per 1,000 days of indwelling catheters was found, whereas no CAI has occurred with the eight microbiologically tested silver catheters.

All 16 centromere DNAs from Saccharomyces cerevisiae show DNA curvature.

All 16 centromere DNA regions of Saccharomyces cerevisiae including 90 bp framing sequences on either side were cloned. These 300 bp long centromere regions were analysed by native polyacrylamide gel electrophoresis and found to display a reduced mobility indicative of DNA curvature. The degree of curvature is centromere dependent. The experimental data were confirmed by computer analysis of the 3-dimensional structure of the CEN DNAs. Altogether these data provide further evidence for a model for budding yeast centromeres in which CEN DNA structure could be important for the assembly, activity and/or regulation of the centromere protein-DNA complex.

IgA1 protease production by bacteria colonizing the upper respiratory tract.

Thirty-eight clinical isolates of Haemophilus influenzae and ten clinical isolates of Streptococcus pneumoniae were examined for IgA1 protease production. A suspension of surface material of each individual strain was incubated with human secretory IgA; IgA1 cleavage products were detected by SDS-PAGE and immunoblotting. The high incidence of IgA1 protease-positive strains (68.4% of the examined H. influenzae and 100% of the examined S. pneumoniae strains) confirms that IgA1 protease activity is a frequent characteristic of these two species. Yet the presence of this enzyme is, if at all, only a minor decisive factor for the induction of symptomatic infections of the upper respiratory tract by IgA1 protease-positive bacteria.

Human 170 kDa and 180 kDa topoisomerases II bind preferentially to curved and left-handed linear DNA.

The binding activities of the 170 kDa and the 180 kDa human topoisomerases II (topo II alpha and topo II beta) to linear DNA fragments with different degrees of curvature were characterized. In gel retardation experiments it was shown that both forms of the enzyme bind preferentially to a curved 287 bp fragment, forming a detectable stable complex. The affinity for straight DNA fragments of similar length is significantly lower. Both a commercially available topo II alpha, isolated from placenta, and topo II alpha and topo II beta purified from nuclear extracts of the Namalwa lymphoma tissue culture line gave similar results. The effects of double-stranded poly[d(A-T)], poly[d(G-C)], supercoiled plasmid DNA and linear Z-DNA on the topo II-complex with curved DNA were analyzed in competition experiments. The hierarchy of affinities of the 180 kDa topo II beta for these DNAs has the order: linear left-handed DNA > supercoiled DNA > or = curved DNA >> poly[d(A-T)] > poly[d(G-C)]. The 170 kDa topo II alpha binds with similar affinity to curved DNA and linear Z-DNA > or = supercoiled DNA >> linear B-DNA. The data imply that human topoisomerase II binding is more sensitive to DNA secondary structure than to DNA sequence per se. The ability of the enzyme to preferentially recognize a wide variety of sequences in unusual secondary structures suggests a mode of targeting the enzyme in vivo to regions of high negative supercoiling.