Efficacy and duration of antistaphylococcal activity comparing three antibiotics bonded to Dacron vascular grafts with a collagen release system.

Type 1 collagen, minimally cross-linked, was used to bind one of three antibiotics (amikacin, chloramphenicol, or rifampin) to double-velour Dacron grafts to develop a prosthesis resistant to infection. Six millimeter disks of graft were placed in separate flasks (specific for each antibiotic) containing albumin in saline and continuously agitated. At daily intervals the solution was changed, and paired graft samples were removed and placed on a blood agar plate confluently streaked with bacteria. The initial zone of inhibition (centimeters squared), the time to 50% reduction of initial inhibition zone, and the overall duration of antibacterial activity were recorded on an exponential model. Grafts bonded with amikacin and chloramphenicol had an overall duration of activity of only 2 and 1 day, respectively, against Staphylococcus aureus. The collagen bonded rifampin grafts had an initial zone of 14.76 cm,2 took 3.92 days to reach 50% of initial inhibition, and had an overall duration of activity of 22.4 days. This was significantly better than grafts preclotted with 1.0 ml of rifampin (60 mg/ml) and 9 ml of blood (10.92 cm,2 1.06 days, and 5.6 days). When tested against a slime-producing Staphylococcus epidermidis (American Type Culture Collection No. 35983), the graft bonded with rifampin had inhibitory activity of up to 27.77 days with a 50% of activity eluted at 4.78 days, significantly better than the preclotted rifampin graft without collagen bonding. These data suggest that rifampin bonded by collagen can protect a vascular graft against infection from S. aureus and S. epidermidis for up to 3 weeks after implantation.     

A passive system using rifampin to create an infection-resistant vascular prosthesis

Vascular prosthesis infection is a devastating complication of vascular operations. The development of a simple process for imparting infection resistance to vascular prosthesis material would be invaluable. Rifampin was added to the blood that was used to preclot 8 mm Dacron vascular prostheses that were used to replace the infrarenal aorta in 10 mongrel dogs. Rings of the grafts were resected after blood had flowed through them for 0 minute, 15 minutes, 60 minutes, and 24 hours. The resected graft rings were placed on culture plates that had been inoculated heavily with Staphylococcus aureus. The rate of antibiotic dialysis from the graft was determined by comparison of the inhibition rings that were produced by the graft rings at each subsequent interval. At 60 minutes, inhibition was 94% of that at time 0. Inhibition at 24 hours was 91%, which demonstrated no significant decrease from 60 minutes. Other antibiotics were screened by this technique, but none of them demonstrated inhibition at 24 hours. In a pilot study in which two dogs were given parenteral rifampin before and after operation and grafts were preclotted with blood that contained rifampin, it was suggested that there was a slight increase in inhibition at 24 hours (93%). The data indicated that rifampin that is added to the blood that is used to preclot a porous Dacron prosthesis is so slowly dialyzed from the graft that inhibition remains at 24 hours. This passive system imparts potential resistance to the prosthesis.     

Use of an antibiotic-bonded graft for in situ reconstruction after prosthetic graft infections.

We have developed an infection resistant vascular prosthesis by bonding rifampin to Dacron grafts with the use of a collagen matrix release system. The purpose of this study was to determine the efficacy of this antibiotic-bonded graft in resisting infection after an in situ reconstruction of a previously infected prosthetic bypass. Eighty-three adult mongrel dogs underwent implantation of a 3 cm untreated Dacron graft into the infrarenal aorta. This initial graft was deliberately infected, at the time of operation, with 10(2) organisms of Staphylococcus aureus by direct inoculation. One week later, the dogs were reexplored, the retroperitoneum debrided, and the animals randomized to undergo an end-to-end in situ graft replacement with either one of two types of prosthetic grafts: group I (collagen, n = 36) received control collagen-impregnated knitted Dacron grafts; group II (rifampin, n = 47) received experimental collagen-rifampin-bonded Dacron grafts. Each group of animals was then subdivided to receive one of four treatment protocols: (a) no antibiotic therapy, (b) cephalosporin peritoneal irrigation solution (cefazolin 500 mg/1000 ml) during operation and two doses of cephalosporin (cefazolin, 500 mg intramuscularly) postoperatively, (c) treatment as in protocol group b plus 1 week of cephalosporin (cefazolin, 500 mg intramuscularly, twice daily), and (d) treatment as in protocol group b plus 2 weeks of cephalosporin (cefazolin, 500 mg intramuscularly, twice daily). All grafts were sterilely removed between 3 and 4 weeks after implantation. There were no anastomotic disruptions and all grafts were patent at the time of removal.(ABSTRACT TRUNCATED AT 250 WORDS)     

Comparative evaluation of the elasticity and flexibility of bioimpregnated knitted grafts

Longitudinal elasticities of whole human blood clot, whole canine blood clot, Factor XIII cross-linked fibrin, glutaraldehyde-fixed human albumin, and formaldehyde-fixed collagen, gelatin and collagen/gelatin were determined and normalized to human whole blood clot. Matrices of a knitted Dacron graft were then impregnated with albumin, collagen, and collagen/gelatin and their longitudinal elasticities were determined and normalized to a preclotted graft. Comparisons were also made for the longitudinal elasticities of a virgin graft, a manipulated control for a preclotted graft, and a manipulated control for the matrix-impregnated grafts. Flexibilities were then calculated based on the weights and elasticities of these grafts and normalized to the flexibility of the preclotted graft. Fibrin had twice and 39 times more longitudinal elasticity than human blood clot and collagen, respectively. The preclotted graft has longitudinal elastic properties similar to a virgin graft, and is 2.8, 2, and 1.4 times more elastic than the albumin, collagen and collagen/gelatin grafts, respectively. The preclotted graft was 2.5, 2, and 1.4 times more flexible than the albumin, collagen and collagen/gelatin grafts, respectively.     

Prevention of graft infection by use of prostheses bonded with a rifampin/collagen release system.

The objective of this study was to test the efficacy of bonding rifampin to double-velour Dacron grafts with collagen to prevent graft sepsis. Fifty 6.0 mm Dacron grafts (length 5.0 cm) impregnated with either collagen (control) or collagen plus rifampin (experimental) were implanted in dogs end-to-end into the infrarenal aorta. The dogs were divided into four groups (each with an experimental and control subdivision) as a function of time between grafting and bacterial challenge. At 2, 7, 10, or 12 days after graft implantation, sequential groups were challenged with 1.2 x 10(8) colony forming units of Staphylococcus aureus (clinical isolate) intravenously suspended in 250 ml normal saline. Three weeks after hematogenous seeding, the grafts were sterilely harvested. One-tailed Fisher's exact test was used to compare the patency and culture-proven infection of control and antibiotic coated grafts as a function of implantation time before bacteremic challenge. In the 2-day group, four of six control grafts were infected compared with zero of six experimental grafts (p less than 0.030). In the 7-day group, five of six control grafts were infected with S. aureus versus zero of six in the experimental group (p less than 0.008). In the 10-day group, one of six experimental grafts was infected, but the control group had only two of six graft infections. In the 12-day group two of six experimental grafts and one of five control grafts were infected. These results indicate that rifampin bonded with collagen to knitted Dacron grafts will protect the graft from bacteremic infection for 7 days after implantation in a highly challenging model.(ABSTRACT TRUNCATED AT 250 WORDS)     

Treatment of established prosthetic vascular graft infection with antibiotics preferentially concentrated in leukocytes

The efficacy of treating established vascular graft infections with rifampin and clindamycin (preferentially concentrated in leukocytes) and cefazolin (not concentrated in leukocytes) was studied in a canine model. Infrarenal aortic, 6 mm by 6 cm knitted Dacron double velour grafts were implanted and infected with 10(8) colony-forming units (CFU) of coagulase-positive Staphyloccus aureus organisms injected intravenously immediately after graft placement. Antibiotic therapy was instituted at 3 months postimplantation. Three groups were studied: (I) untreated controls (n = 3); (II) therapy with intravenous cefazolin 15 mg/kg/8 hr for 28 days (n = 7); and (III) combined therapy with intravenous rifampin 13 mg/kg/24 hr and intravenous clindamycin 13 mg/kg/8 hr for 28 days (n = 7). Grafts were removed for quantitative bacteriologic studies after the 28-day course of therapy. Two group I control grafts remained patent with 6.4 X 10(6) and 8.1 X 10(3) CFU S. aureus/gm of graft. The third control graft was thrombosed. Two group II animals demonstrated 1.6 X 10(7) and 2.3 X 10(5) CFU S. aureus organisms/gram of graft, respectively; the remaining five group II grafts were free of organisms. All group III grafts were sterile--a significant difference (p less than 0.05) from group I grafts. In this experimental model, established prosthetic graft infections were eradicated by intensive treatment with antibiotics preferentially concentrated in leukocytes.     

Prevention of prosthetic vascular graft infection by rifampicin impregnation of a protein-sealed Dacron graft in combination with parenteral cephalosporin.

Antibiotic-impregnated prosthetic vascular grafts have been shown to be highly resistant to bacterial contamination in animal models. The aim of this study was to assess if graft infection in an animal model challenged with local bacterial contamination could be reduced further by the use of rifampicin soaking of a protein-sealed Dacron graft in addition to the prophylaxis provided by perioperative intravenous cephalosporin. Of 20 Merino sheep given cefoxitin intravenously before the induction of anaesthesia, ten had a rifampicin treated Dacron graft implanted into the common carotid artery (group 1), and ten received an untreated Dacron graft (group 2). Before wound closure the grafts were inoculated with 1 ml of 10(8) colony forming units per ml of Staphylococcus aureus. After three weeks the grafts were removed and cultured. Group 1 sheep had fewer graft infections (two of 10) compared to group 2 (six of eight survivors), this difference being statistically significant (p = 0.03; Fisher exact test). It is concluded that rifampicin treatment of protein-sealed Dacron grafts combined with intravenous cefoxitin provides more protection from contaminating Staphylococcus aureus than intravenous cefoxitin alone in this animal model.     

Antibacterial activity,antibiotic retention,and infection resistance of a rifampin-impregnated gelatin-sealed Dacron graft

Purpose: A gelatin-sealed porous Dacron graft impregnated with rifampin was evaluated in a two-part study of its use in preventing prosthetic infection.Methods: The graft was impregnated by soaking it for 15 minutes in rifampin (1 mg/ml). In part 1 its antibacterial activity and rifampin retention over time were determined. Infrarenal aortic replacement was performed in pigs, and the rifampin concentration of the graft, serum, and perigraft space was assayed up to 96 hours after surgery. In part 2, infection resistance was tested in pigs in which the retroperitoneum was contaminated with Staphylococcus aureus after graft replacement. The postoperative infection rate was compared in three groups: pigs given gelatin-sealed grafts without rifampin (controls), pigs receiving nonimpregnated grafts and intravenous rifampin (15 mg/kg) for 3 days after surgery, and those given the rifampin grafts.Results: Rifampin was present in the grafts for up to 72 hours after surgery and in the perigraft fluid for 24 hours but was never detected in the serum. The grafts had inhibitory activity in vitro against S. aureus and the biofilm phase of Staphylococcus epidermidis for up to 3 days and against Escherichia coli for 2 days. Pigs given intravenous rifampin had a significantly lower infection rate than had control pigs (7/12 vs 13/13; p = 0.02); those receiving the rifampin graft had a lower rate (2/13) than had either the control pigs (p < 0.001) or those given intravenous rifampin (p < 0.04).Conclusions: This simple method of graft impregnation resulted in antibiotic retention for 3 days and appeared to be superior to intravenous antibiotic administration in preventing perioperative graft infection. (J VASC SURG 1994;19:675-82.)     

Simple methods for direct antibiotic protection of synthetic vascular grafts

Two simple methods for direct antibacterial protection of synthetic vascular grafts were investigated. In the first protocol the highly protein-bound antibiotics nafcillin (90% protein bound), cefazolin (80%), and cefamandole (70%) were added directly to preclotting blood. Knitted Dacron grafts preclotted in the presence of one of these drugs absorbed significant amounts. Although at high concentrations these antibiotics exhibited anticoagulant effects, significant antibacterial protection was obtained at lower antibiotic levels. Washing treated grafts for 6 hours failed to eliminate the antibacterial activity. Antibiotics remained on the grafts for at least 96 hours. In the second protocol knitted Dacron grafts were soaked in a suspension of silver-pefloxacin, a silver-nalidixic acid analogue with intense antistaphylococcic activity. Using 110Ag-labeled complexes, significant antibiotic activity was documented on the graft after 19 days of washing. Four nafcillin-treated prostheses, six silver-pefloxacin-coated grafts, and 11 control grafts were interposed in the infrarenal aorta of dogs and immediately challenged with an intravenous infusion of 1 X 10(7) Staphylococcus aureus. None of the four nafcillin-treated grafts was infected at 3 weeks. One of the six silver-pefloxacin-coated grafts grew staphylococci, and 9 of 11 controls had positive graft cultures for Staphylococcus when harvested. These studies suggest that prosthetic grafts can be simply coated at the time of implantation with antibiotics selected for appropriate binding and antibacterial characteristics to obtain an infection-resistant prosthesis.     

The influence of early surface thromboreactivity on long-term arterial graft patency

The influence of early graft surface thromboreactivity on long-term arterial polyester (Dacron) graft patency was investigated with separate ex vivo and in vivo animal models. First, parallel, flow-regulated external aortocaval fistulae were created in five pigs with use of paired 8 mm X 35 cm crimped, warp-knitted, low-profile filamentous velour Dacron tubes: one tube preclotted with autologous blood, the other autoclaved after being soaked in human albumin. Autologous radiolabeled platelets, red cells, and radiolabeled human fibrinogen were injected at initiation of graft flow, with timed graft samples submitted for isotope gamma well-counting. Flow surface accumulation of radiolabeled blood elements was greater on the preclotted graft limb at-all time intervals studied, greatest after 5 minutes of flow initiation with RBC accumulation on the preclotted limb 5.19 +/- 0.84 (x +/- S.E.), platelet accumulation 5.57 +/- 1.00, and fibrinogen accumulation 1.82 +/- 0.14 times greater than that on the albumin-treated limb. Second, bilateral iliofemoral artery bypass grafts were placed in 12 mongrel dogs using 6 mm X 10 cm externally supported, noncrimped, warp-knitted, low-profile filamentous Dacron tubes. Before implant in each dog, one graft limb was clotted with autologous blood and the other was autoclaved after being soaked in 25% human albumin. Fresh autologous radiolabeled platelets were injected after wound closure in seven of these dogs. Postimplant graft imaging at 24 and 72 hours showed radiolabeled platelet accumulation to be 1.43 +/- 0.21 and 2.05 +/- 0.18 times greater on the preclotted graft limb. Six of 12 preclotted graft limbs and 7 of 12 albumin-treated graft limbs were patent when animals were killed 5 to 6 months after implant (not significant). Heat-denatured albumin-coated Dacron surfaces have a reduced early thromboreactivity but do not appear to greatly potentiate long-term arterial graft patency.     

Accelerated Healing of Dacron Grafts Seeded by Preclotting with Autologous Bone Marrow Blood

> Studies have suggested that bone marrow-derived cells in the circulation may have the capacity and potential to endothelialize and heal vascular graft surfaces. We have investigated whether accelerated endothelialization could be achieved for Dacron grafts seeded by preclotting with bone marrow blood (BMB). Five 8 mm x 6 cm Dacron grafts seeded and preclotted with BMB and four controls preclotted with peripheral blood were implanted in the descending thoracic aorta (DTA) of mongrel dogs for 2 and 4 weeks. Two additional BMB DTA grafts were studied for 3 months. Five pairs of BMB and control grafts (4 mm x 6 cm) were bilaterally implanted into the carotids of dogs for 1 week and five pairs for 4 weeks. All grafts remained patent. BMB seeding/preclotting was a simple, effective method to accelerate early graft endothelialization without increasing thrombogenicity. Further studies are needed before clinical application can be recommended.     

Prevention of vascular graft infection by rifampin bonding to a gelatin-sealed dacron graft

This study examines the efficacy of rifampin bonding to a gelatin-sealed knitted Dacron graft to prevent perioperative bacteremic vascular graft infection. Antibiotic bonding was obtained by soaking grafts for 15 minutes in a 1 mg/ml saline solution of rifampin at 37°C. Nineteen dogs had thoracoabdominal aortic bypass: seven (group I) received a rifampin treated graft; six (group II) received an untreated gelatin-coated graft; and six (group III) received an uncoated Dacron graft. Two days later bacteremic challenge was produced by rapid intravenous injection of 5×10 ⁵ colony forming units of methicillin resistantStaphylococcus aureus.Grafts were harvested five days after this challenge and cut into 10 fragments, each submitted to bacterial counts. Results were expressed as CFU/cm ² of graft material. In group I, no graft was infected, whereas all grafts in groups II and III were infected (p<0.05). Median bacterial counts from the infected fragments (median±SD) were similar in groups II (2.5×10⁵ CFU/cm²) and III (4×10⁴ CFU/cm²). Blood cultures at time of sacrifice were negative in all dogs in group I and positive in five of six dogs in groups II and III. Cultures of liver, spleen, kidney, and lung specimens were always negative in group I and positive in 22 of 24 specimens in group II and 23 of 24 specimens in group III. Soaking a gelatin-sealed Dacron graft in rifampin solution evidently prevents early bacteremic graft infection and secondary foci of infection in this model.Presented at the Annual Meeting of the French Vascular Surgery Society, Nancy, France, May 18–19, 1990.     

Rifampin and Triclosan but not silver is effective in preventing bacterial infection of vascular dacron graft material.

OBJECTIVES: To evaluate the efficacy of silver- or Triclosan-coated prosthetic material compared to Rifampin bonded Dacron concerning their resistance to infection following subcutaneous implantation and contamination with Staphylococcus aureus. DESIGN: Animal experimental study in mice. MATERIAL AND METHODS: Thirty-six C3H/HcN mice (Charles River Lab., Sulzfeld, Germany) with a weight between 24 and 27 g were randomised into six groups counting six animals each. Group I: control, gel-sealed dacron graft, group II: gel-sealed dacron graft and local contamination, group III: Intergard-Silver-prosthesis and contamination, group IV: silver/gel-sealed dacron prosthesis (test graft) and contamination, group V: Rifampin-bonded gel-sealed graft and contamination, group VI: Triclosan/collagen-coated dacron graft and contamination. Dacron graft material 0.8x1 cm was subcutaneously implanted in mice. Local contamination with 2x10(7)/0.2 ml S. aureus ATCC 25923 was carried out in groups II to VI. On day 14 the animals were killed and the grafts were explanted. The microscopic, histologic and microbiological evaluation of the graft material and the perigraft tissue was performed. RESULTS: In control group I no case of infection was detected. In group II, 6 of 6 animals showed infection. In group III (Intergard-Silver) and group IV (silver/gel-test graft) were 6 of 6, in group V (Rifampin) only 1 of 6 grafts and in group VI (Triclosan) 4 of 6 grafts were infected. The difference between the low rate of infection in group V (Rifampin) in comparison to the completely infected groups III and IV (Silver) as well as the control group II was significant. Treatment of grafts with Triclosan could prevent infection only in 1/3 of the cases in group IV. CONCLUSION: Silver coating failed to prevent graft infection material. A potential antimicrobial property was evident for Triclosan whereas Rifampin-bonded grafts exhibit a significantly reduced infection rate. Thus, silver-coated vascular grafts cannot ensure protection from vascular graft infection.     

Prophylaxis against Staphylococcus aureus vascular graft infection with mupirocin-soaked, collagen-sealed dacron

A rat model was used to investigate the efficacy of mupirocin in the prevention of vascular prosthetic graft infections. The effect of mupirocin-soaked Dacron was compared with the effect of rifampin-soaked, collagen-sealed Dacron in the rat model of graft infection caused by methicillin-susceptible Staphylococcus aureus and methicillin-resistant S. aureus. Graft infections were established in the back subcutaneous tissue of 195 adult male Wistar rats by implantation of 1-cm(2) Dacron prostheses followed by topical inoculation with 5 x 10(7) colony-forming units of S. aureus. The study included a control group (no graft contamination), two contaminated groups that did not receive any antibiotic prophylaxis, two contaminated groups in which perioperative intraperitoneal amoxicillin clavulanate prophylaxis (50 mg/kg) was administered, four contaminated groups that received mupirocin- or rifampin-soaked graft, and four contaminated groups that received mupirocin- or rifampin-soaked graft and perioperative intraperitoneal amoxicillin clavulanate prophylaxis (50 mg/kg). The grafts were sterilely removed 7 days after implantation and the infection was evaluated by using sonication and quantitative agar culture. Data analysis showed that the efficacy of mupirocin against both strains was significantly different from that of the untreated control. In addition, mupirocin was more effective than rifampin against the methicillin-resistant strain. Finally, only the combination of mupirocin and amoxicillin clavulanate produced complete suppression of growth of all strains.     

Experimental examination concerning the efficacy of silver-coated Dacron prostheses in vascular graft infections following subcutaneous implantation in a standardized infection model

It was the aim of the study to examine the efficacy of silver coated prostheses in comparison to Rifampin in impregnated prostheses in the prevention of vascular graft infections. MATERIAL AND METHODS: 24 C3H/HcN mice with a bodyweight between 24 and 27 grams were assigned to four different groups. GROUP I: control gel-sealed Dacron graft (Uni-Graft DV) (6), GROUP II: gel-sealed Dacron graft (Uni-Graft DV) contaminated locally with 2 x 10(7) CFU/1.2 ml Staphylococcus aureus ATCC 25923 (6), GROUP III: silver prosthesis (Intergard Silver) contaminated locally with 2 x 10(7) CFU/0.2 ml Staphylococcus aureus ATCC 25923 (6), GROUP IV: Rifampin impregnated prosthesis contaminated locally with 2 x 10(7) CFU/0.2 ml Staphylococcus aureus ATCC 25923 (6). 14 days after primary operation all animals were euthanized and the grafts harvested. Specimens were examined for signs of infections by histology and microbiology. RESULTS: At termination of the trial on day 14 none of the grafts of group I were contaminated. 6 out of 6 grafts in group II, 6 out of 6 grafts in group III and 1 out of 6 grafts in group IV presented with infected grafts. The use of antimicrobial Rifampin could significantly prevent infection after bacterial challenge in group IV. CONCLUSION: The silver protected prosthesis (Intergard Silver) seems to be not effective in protecting vascular infection in vivo. However, the Rifampin group showed excellent results. In conclusion Rifampin bonded gelatin-sealed Dacron grafts are significantly more resistant to bacteremic infection than are silver/collagen-coated Dacron grafts.     

Preincubation of Dacron grafts with recombinant tissue factor pathway inhibitor decreases their thrombogenicity in vivo

Background: We have previously shown that preincubation of whole blood clots with recombinant tissue factor pathway inhibitor (rTFPI) attenuates clot-associated procoagulant activity assessed ex vivo. This study was undertaken to determine whether a single local application of rTFPI induces similar attenuation of the procoagulant activity on preclotted Dacron grafts implanted in an artery in vivo.Methods: Dacron grafts (4 mm × 4 cm long) were preclotted in porcine blood and incubated with either rTFPI (5 mg/ml) or arginine-phosphate buffer for 15 minutes. Grafts were implanted end-to-end in the femoral arteries of 10 pigs, with one rTFPI-treated and one buffer-treated graft implanted in each animal. Animals did not undergo anticoagulation either before or after graft implantation. Radiolabeled porcine fibrinogen was injected intravenously, and the grafts underwent perfusion for 1 hour. A subgroup of animals (n = 7) also had infusion of radiolabeled autologous platelets at the time of administration of radiolabeled fibrinogen.Results: Fibrin(ogen) deposition was decreased in rTFPI-treated grafts by 36% ± 7% (mean + SEM) compared with buffer-treated grafts (p = 0.001). Platelet deposition was also reduced in the rTFPI-treated grafts by 31% ± 15%, although the reduction did not reach statistical significance (p = 0.10). The extent of rTFPI-mediated attenuation of fibrin(ogen) versus platelet deposition varied independently among animals.Conclusions: Clot-directed anticoagulant effects of rTFPI appear to be useful for substantially decreasing the thrombogenicity of Dacron grafts immediately after their implantation. Chronic studies to determine whether the decreases in thrombogenicity result in improved long-term graft patency appear warranted.(J Vasc Surg 1996;24:865-70.)     

Infection of vascular prostheses caused by bacterial biofilms

A canine model was developed to study the efficacy of graft replacement as treatment for vascular prosthesis infections from Staphylococcus epidermidis. Infrarenal aortic graft infections were established in 18 dogs by implantation of Dacron prostheses colonized in vitro with a slime-producing strain of S. epidermidis to form an adherent bacteria-laden biofilm (5 X 10(6) colony-forming units/cm2 graft). Study animals developed a graft infection with anatomic and microbiologic characteristics typical of late prosthetic graft infections in humans (sterile perigraft exudate, absent graft incorporation, and normal serum leukocyte count and sedimentation rate). The S. epidermidis study strain was isolated from 14 of 18 explanted grafts (78%) by mechanical disruption of the graft surface biofilm and culture in broth media. Four dogs with sterile graft cultures had histologic evidence of bacterial infection. The established prosthetic surface biofilm infection was treated by graft excision, parenteral cefazolin, and graft replacement with a Dacron or polytetrafluoroethylene (PTFE) vascular prosthesis. One month after graft replacement, no PTFE graft had signs of infection, but perigraft exudate and inflammation involved three of nine Dacron grafts (33%). The study strain was recovered from four of nine PTFE grafts (44%) and two of nine Dacron (22%) replacement grafts (p greater than 0.05). Prosthetic replacement of Dacron prostheses infected by S. epidermidis as a bacteria-laden surface biofilm can result in early graft healing, but persistent colonization of one third of replacement grafts signify that recurrent clinical infection remains a risk.     

Impact of postantibiotic effect on bacterial adherence to vascular prostheses

The brief exposure of bacteria to high antibiotic concentrations can result in prolonged suppression of bacterial growth termed postantibiotic effect (PAE). A pathogenic Staphylococcus epidermidis strain (RP-62) was exposed to 2X and 6X minimum inhibitory concentrations (MIC) of cefazolin, vancomycin, or rifampin for 1 hr and incubated for 2 to 24 hr in inhibitory-free broth. PAE was defined as the difference in time required for a 1.0 log increase in test (T) vs control (C) cultures (PAE = T-C). PAE was observed only for rifampin: 6 hr at 2X MIC and 8 hr at 6X MIC. Bacterial adherence to Dacron grafts was calculated in PAE vs control cultures by a quantitative culture technique for graft specimens incubated 2 to 24 hr. A demonstrable PAE and its impact on adherence were found to be both antimicrobial and concentration dependent. A significant decrease in staphylococcal adherence to velour-knitted Dacron was demonstrated by rifampin at 2X MIC (P less than 0.05) and 6X MIC (P less than 0.01). This phenomena may be useful in reducing bacterial adherence and colonization of bioprosthetics in the perioperative period. Preoperative suppression of staphylococcal skin flora by high dose antimicrobials can alter the capacity of these organisms to adhere to vascular prosthetic grafts and, if incorporated into antibiotic prophylaxis regimens, may reduce graft colonization.     

Temporin A as a prophylactic agent against methicillin sodium-susceptible and methicillin sodium-resistant Staphylococcus epidermidis vascular graft infection

OBJECTIVE: The purpose of this study was to investigate the efficacy of temporin A as a prophylactic agent in a rat model of vascular graft infection from methicillin sodium-susceptible and methicillin sodium-resistant Staphylococcus epidermidis. METHODS: The prospective, randomized, controlled animal study set in a research laboratory in a university hospital used 280 adult male Wistar rats (weight range, 280 to 350 g). Graft infections were established in the back subcutaneous tissue of rats with implantation of 1-cm(2) sterile Dacron grafts followed by topical inoculation with 2 x 10(7) colony-forming units of S epidermidis. The study for each staphylococcal strain included: one control group (no graft contamination), one contaminated group that did not receive any antibiotic prophylaxis, one contaminated group that received temporin A-soaked graft, two contaminated groups that received perioperative intraperitoneal cefazolin (30 mg/kg) or vancomycin hydrochloride prophylaxis (10 mg/kg), and two contaminated groups that received temporin A-soaked graft and perioperative intraperitoneal cefazolin (30 mg/kg) or vancomycin hydrochloride (10 mg/kg) prophylaxis. All grafts were explanted at 7 days after implantation. The main outcome measure was quantification of bacterial contamination. RESULTS: Overall, the perioperative prophylaxis based on soaked grafts was not significantly different to that of parenteral vancomycin hydrochloride. Only the combination between temporin A and vancomycin hydrochloride produced a complete bacterial inhibition for both strains. CONCLUSION: Temporin A showed a similar antibacterial in vitro activity against the two different strains. The in vivo results suggest its potential use in providing prophylaxis to direct graft contamination when used in combination with parenteral vancomycin hydrochloride.     

Inhibition of vascular prosthetic graft infection using a photocrosslinkable chitosan hydrogel

BACKGROUND: Despite improvements in surgical techniques and antimicrobial therapies, prosthetic aortic graft infections remain a clinical problem. It is well known that chitosan has strong antibacterial activities to a wide variety of bacteria including Staphylococcus aureus, epidermidis and Escherichia coli (E. coli). The antibacterial activity by adhering a photocrosslinkable chitosan hydrogel to Dacron grafts was investigated in vitro and in vivo using a rabbit model. MATERIALS AND METHODS: The photocrosslinkable chitosan hydrogel (50microl) coated grafts (3 x 2mm fragments) were evaluated on a resistance against E. coliin vitro. The graft infections in vivo were also initiated through implantation of a Dacron graft fragment into the infrarenal aorta of a rabbit, followed by a topical inoculation with 10(6) colony-forming units of E. coli. The graft infection was allowed to develop over the following 1 week. RESULTS: The photocrosslinkable chitosan hydrogel-coated grafts exhibited a resistance against E. coliin vitro. Furthermore, application of 0.1ml photocrosslinkable chitosan hydrogel on the Dacron implant in vivo substantially inhibited graft infection with E. coli. CONCLUSIONS: These preliminary results suggested the potential use of a photocrosslinkable chitosan hydrogel in directing graft infection prophylaxis.