Lester Sampath

Decreased bacterial adherence and biofilm formation on chlorhexidine and silver sulfadiazine-impregnated central venous catheters implanted in swine.

OBJECTIVE To determine if antiseptic central venous catheters impregnated with silver sulfadiazine and chlorhexidine (antiseptic) reduce bacterial adherence and biofilm formation without producing local or systemic toxicity. DESIGN Prospective, randomized, controlled trial. SETTING Experimental laboratory in a university teaching hospital. SUBJECTS Ten outbred New Hampshire pigs. INTERVENTIONS Nonimpregnated (control) and antiseptic-impregnated catheters were inserted intravascularly into swine for 7 days. After explantation, the catheters were assessed for bacterial adherence and biofilm formation, and the surrounding tissue was assessed for signs of toxicity. Before retrieval, systemic concentrations of antimicrobials were determined. MEASUREMENTS AND MAIN RESULTS Sequential roll plate and centrifuging were used to detect moderately and tightly adherent bacteria on the outer and luminal surfaces of the catheter. The presence of biofilm was detected by scanning electron microscopy. Tissues surrounding the catheters were examined histopathologically; systemic concentrations of chlorhexidine, sulfadiazine, and silver were determined by atomic absorption and high-performance liquid chromatography. As compared with the controls, antiseptic catheters had significantly (p < .01) fewer moderately and tightly adherent bacteria on outer and luminal surfaces, and fewer adherent bacteria when outer surfaces alone were examined (p < .01). Scanning electron microscopy showed bacterial biofilm and adherence on the control catheters but not on the antiseptic catheters. There were no abnormal histopathologic changes associated with the test catheter, and serum concentrations of the antibacterial agents were shown to be within nontoxic ranges. CONCLUSION The antiseptic-impregnated catheters prevented bacterial adherence and biofilm formation and produced no local or systemic toxicity.

Development of an infection-resistant LVAD driveline: a novel approach to the prevention of device-related infections

BACKGROUND Infection remains the single most important challenge to extended left ventricular assist device (LVAD) use and often arises from the percutaneous driveline exit site. We evaluated the ability of an LVAD driveline prototype impregnated with chlorhexidine, triclosan, and silver sulfadiazine to resist bacterial and fungal colonization. METHODS The spectrum and duration of antimicrobial activity were evaluated in vitro by daily transfer of driveline segments embedded on agar plates inoculated with 10(8) colony-forming units (CFU) of Staphylococcus aureus (S. aureus), Staphlococcus epidermidis, Enterobacter aerogenes, Psuedomonas aeruginosa, and Candida albicans, and then measuring zones of inhibition around the sample subsequent to 24 hours of incubation at 37 degrees C. Antimicrobial activity was demonstrated against all organisms for greater than 14 days, and for over 21 days for gram-positive bacteria. To demonstrate in vivo efficacy of the treated driveline, 3-cm segments of driveline were implanted in the dorsal and ventral surface of rats. The exit site was inoculated with 10(6) CFU of S. aureus. After 7 days, driveline segments were aseptically explanted and assayed for bacterial colonization and retention of antimicrobial activity. One hundred percent of control segments were colonized (10(5) CFU S. aureus/cm) as against 13% of the test explants (< or = 330 CFU/cm; p < 0.0001). RESULTS Subcultures of the insertion site and driveline pocket tissue resulted in 10(3) to 10(5) CFU per swab culture for control rats and 0 to 10(2) CFU/swab for test animals. Test drivelines retained 80% of anti-S. aureus activity. Gross and histological examination of the driveline and surrounding pocket revealed minimal tissue reactivity with positive signs of tissue ingrowth. CONCLUSION An antimicrobial driveline may prevent early infections and facilitate ingrowth of tissue to provide long-term stability and protection against late infection.

Control of prosthetic bacterial infection: Evaluation of an easily incorporated, tightly bound, silver antibiotic PTFE graft

Despite the use of prophylactic antibiotics in vascular surgery, prosthetic infection rate remains 2-5%. Antibiotics bound to vascular prostheses can control experimentally induced infection but prolonged antibacterial activity has not been achieved. This study evaluates the in vivo efficacy and antibiotic retention of an easily prepared silver-antibiotic prosthesis. Prostheses were prepared by combining silver with oxacillin or amikacin using an organic solvent. After evaporation of the solvent, the graft was left impregnated with the antibiotic complex. In vivo retention studies were performed by implanting PTFE 110Ag-oxacillin prostheses in four canine abdominal aortas. When prostheses were explanted at 1 week, mean antibiotic retention was found to be 20% of original activity, higher than the mean inhibitory concentration for Staphylococcus aureus. In three groups of five dogs, 20 X 7-mm prostheses of PTFE alone, PTFE silver-oxacillin, or PTFE silver-amikacin were implanted in the abdominal aorta and the grafts were inoculated with 10(7) S. aureus of a known bacteriophage type, in a closed retroperitoneal pocket. The animals were sacrificed at 1 week and the prostheses were excised for quantitative bacterial culture. PFTE silver-oxacillin, and PTFE silver-amikacin prostheses had 1.7 X 10(2) and 2.0 X 10(2) colonies, respectively, significantly less (P less than 0.05) than controls (1.3 X 10(6) colonies). These data suggest that antibiotic prostheses can be easily prepared without binding agents. They retain the bound antibiotic for a prolonged period and are effective in reducing graft infection in a stringent direct contamination model.

Combined topical use of silver sulfadiazine and antibiotics as a possible solution to bacterial resistance in burn wounds.

The superior efficacy of quinolones (norfloxacin, pefloxacin, and enoxacin) in controlling burn wound infections signals the discovery of new topical agents. However, there are a few reports on the emergence of resistant mutants to quinolones. Since attempts to develop AgSD resistant strains in vitro were unsuccessful and the emergence of AgSD resistance in vivo is a rare occurrence, we decided to investigate if the combined use of AgSD with other effective antibiotics, especially quinolones, would minimize the development of resistant bacteria. Our in vitro results indicate that when Ps. aeruginosa cultures were serially transferred 10 times through subinhibitory concentrations of norfloxacin, pefloxacin, etc., the MIC increased 40 times while when the cultures were passed through a combination of AgSD and these quinolones, the MIC of quinolones increased only tenfold. In vivo, when burned mice infected with either AgSD sensitive or resistant Ps. aeruginosa strains were treated with a topical cream containing 10mM silver sulfadiazine and 5mM norfloxacin or 5mM pefloxacin, the mortality was much lower than that of 10mM silver sulfadiazine alone or 5mM quinolones alone. Thus, combined use of silver sulfadiazine and quinolones appears to diminish the ability of Ps. aeruginosa strains to form resistant mutants. Furthermore, when the combination is used as a topical agent in burn wounds, lesser amounts of the individual drug are needed to control infection thereby reducing the toxic effects, if any, associated with these drugs. This combination does not in any way interfere with the antifungal or antibacterial properties of these individual drugs.

PTFE graft treated with silver norfloxacin (AgNF): Drug retention and resistance to bacterial challenge

Norfloxacin (NF) and silver norfloxacin (AgNF) were used to coat polytetrafluoroethylene (PTFE) vascular prostheses by using triododecylmethylammonium chloride as a cationic surfactant. The relative retention of the drug on the graft after subjecting it to the biological environment for 3 weeks and antibacterial activity against coagulase-positive Staphylococcus aureus and Escherichia coli were determined. In addition, the ability of AgNF-coated PTFE grafts to sterilize perigraft tissue after mixed bacterial contamination was studied and compared with control PTFE grafts in 10 dogs. At the end of 21 days, 15 and 18% of AgNF were retained on the grafts tested in in vitro and in vivo experiments, respectively. During the same time AgNF-coated grafts from in vitro experiments exhibited significant antibacterial activity (25 mm zone of inhibition (z.i.)), but antibacterial activity was negligible (10 mm z.i.) in the grafts from in vivo experiments. NF retention on the grafts could not be determined because of spectral interference by blood. The antibacterial activity of NF-coated grafts significantly declined after 24 hr in in vivo experiments, hence further evaluation of NF-coated grafts was not done. Seven perigraft sites surrounding AgNF-coated grafts were sterile, but only one perigraft site surrounding control grafts was sterile (P less than 0.05). Cultures from six of nine perigraft infections surrounding control grafts yielded heavy bacterial growth. There was only one wound infection at the site of AgNF graft while there were seven severe wound infections at control graft sites. AgNF-coated grafts exhibited prolonged antibacterial activity compared to NF-coated grafts and offered significant protection against infection from local bacterial contamination.

A topical cream containing a zinc gel (allergy guard) as a prophylactic against latex glove-related contact dermatitis.

Background: Many health care workers are sensitized to the proteins in natural rubber latex and get contact dermatitis as a result of wearing latex gloves. Objective: The aim of the study was to evaluate a topical formulation containing a zinc gel (Allergy Guard) as a prophylactic against latex glove‐related contact dermatitis. Methods: The study was carried out with volunteers who exhibited mild to moderate contact dermatitis (type IV) after wearing latex gloves as per the protocol. Results: Allergy Guard significantly reduced skin irritation in volunteers who exhibited type IV hypersensitivity when exposed to latex gloves. Allergy Guard also exhibited a barrier effect as shown by the permeation of chlorophyllin dye into the skin of volunteers. Allergy Guard prevented dermal irritation induced by sodium lauryl sulfate. Conclusion: Topical formulations containing a zinc gel may be used to delay or prevent latex sensitivity, especially among health care professionals.

Rapid Inactivation of Infectious Pathogens by Chlorhexidine-Coated Gloves

OBJECTIVE Gloves containing chlorhexidine gluconate in an instant-release matrix on their inner surface (CHG gloves) were tested to determine their ability to rapidly inactivate infectious pathogens that may permeate or leak through the latex surface. DESIGN CHG gloves were exposed for 1 to 10 minutes to blood or media containing infectious pathogens (e.g., bacteria, fungi, parasites, and viruses) as well as to lymphocytes and macrophages that are known to be the primary carriers of human immunodeficiency virus (HIV). Inactivation of pathogens was determined either by in vitro assay or in vivo infectivity. Stressed control and CHG glove fingers were submerged in a viral pool (retrovirus or bacteriophage) and after a set time, the glove interiors were checked for presence of permeated virions. RESULTS CHG gloves rapidly inactivate all the pathogens tested including retrovirus and hepatitis B virus (90% to 100%). In the stressed glove fingers, live virus was detected in 26% of the control group but not in any of the CHG group. CONCLUSIONS The use of CHG gloves may reduce the risk of exposure to infectious fluid-borne pathogens should the integrity of the latex barrier be compromised by overt failure or by permeation of viruses. Rapid destruction of lymphocytes and macrophages may facilitate inactivation of HIV associated with these cells. Tests have shown that CHG coating does not alter physical properties of the glove, and, furthermore, CHG gloves do not show potential for dermal irritation or sensitization.

Safety and efficacy of an improved antiseptic catheter impregnated intraluminally with chlorhexidine.

The safety and efficacy of a second-generation improved antiseptic catheter impregnated with silver sulfadiazine and increased levels of chlorhexidine on its outer surface and chlorhexidine alone on its luminal surfaces was compared in vitro and in vivo to standard antiseptic catheters impregnated with these antimicrobials on their outer surfaces only. In rat and pig intravenous models, the improved antiseptic catheter was significantly more effective in resisting both outer surface and luminal colonization compared with the standard antiseptic or control catheters. There was no evidence of tissue toxicity in any group.