JOURNAL
OF SURGICAL
RESEARCH
50,175-178
Antibacterial
(1991)
Effects of Fibrin Glue-Antibiotic
HARRY B. KRAM, M.D.,2 MAHDUBANSAL, Departments
of Surgery and Infectious and the UCLA School Submitted
PH.D.,
Mixtures’
OMATIMBERLAKE,ANDWILLIAMC.SHOEMAKER,
Disease, Los Angeles County King/Drew Medical of Medicine, Los Angeles, California 90024 for publication
February
M.D.
Center,
20, 1990
hemostasis and sealing anastomoses against leakage [l71. The adhesive is slowly dissolved over time by the body’s fibrinolytic system and is completely removed by macrophages within 2 to 4 weeks; normal wound healing progresses during this time. The combination of FG and antibiotics (Abs) has potential clinical applications in the sealing of gastrointestinal, vascular, and tracheobronchial anastomoses, as well as in the establishment of hemostasis in potentially contaminated or infected wounds. Ciprofloxacin and Teicoplanin are two newly developed Abs which are highly effective against a broad spectrum of bacteria. In the present in vitro study, we investigated the duration of action and antibacterial effects of various combined FG-Ab mixtures including Ciprofloxacin, Teicoplanin, Cefoxitin, and Gentamicin. In addition, the effect of FG alone on bacterial growth was evaluated.
In the present in vitro study, we investigated the duration of action and antibacterial effects of nonautologous fibrin glue (FG) combined with antibiotics (Abs) including Ciprofloxacin, Teicoplanin, Cefoxitin, and Gentamicin; the effect of FG alone on bacterial growth was also evaluated. The rate of Ab diffusion from combined FG-Ab clots was evaluated by separate elution with pooled human serum (HS) and normal saline (NS); supernatants were removed daily and assayed for active concentrations of each Ab. The effects of FG and combined FG-Ab clots on bacterial growth were evaluated by inoculating brain-heart infusion (BHI) with Staphylococcus aureus, followed by the addition of FG or FG-Ab clots; a separate set of studies was also performed with the addition of mouse lung homogenate (MLH) as well. The addition of Ab to FG clots resulted in continuous diffusion of the Ab into the surrounding HS or NS for up to 5 to 7 days; however, more than two-thirds of the Ab diffused out within 2 to 3 days regardless of the Ab used. The antibacterial effects of FG-Ciprofloxacin and FG-Teicoplanin clots were significant compared to those of FG clots not containing Ab. In addition, the presence of FG clots (in BHI) resulted in a reduction in bacterial growth compared to that of BHI alone. The addition of MLH to BHI resulted in increased bacterial growth, but this effect was inhibited by the presence of combined FG-Ab clots. In conclusion: (1) a variety of Abs can be combined with FG resulting in significant antibacterial effects in vitro, and (2) the presence of nonautologous FG alone inhibits S. aureus growth in vitro. Potential applications of FGAb mixtures include sealing of gastrointestinal, vascular, and tracheobronchial anastomoses, and the establishment of hemostasis in potentially contaminated or infected wounds. 0 1991 Academic Press, Inc.
MATERIALS Fibrin
AND METHODS
Glue
The FG (Immuno AG; Vienna, Austria) used in the study consisted of highly concentrated human fibrinogen, thrombin (500 US [NIH] units3/ml), calcium chloride (40 pm/ml), and aprotinin (3000 KIU*/ml). After preheating to 37”C, the fibrinogen was dissolved in aprotinin and the dried thrombin reconstituted in calcium chloride, with or without Ab. The components were drawn up into separate syringes which were connected to a single port that allowed simultaneous dispension and mixing of the components. The reconstituted adhesive contained 70 to 100 mg/ml of fibrinogen, 2 to 7 mg/ml of fibronectin (Cold Insoluble Globulin), 10 units5/ml of Factor XIII, and 35 pg/ml of plasminogen. Experimental
Protocol
Fibrin glue (FG), made with highly concentrated human fibrinogen and clotting factors, is a biologic adhesive that has been found to be effective in establishing
One milliliter FG-Ab clots containing 20 pg/ml of Ab were prepared and allowed to solidify at the bottom of
i Presented at the 15th Annual Meeting of the Society for Biomaterials, Lake Buena Vista, FL, April 28-May 2, 1989. ’ To whom reprint requests should be addressed at Department of Surgery, LA County King/Drew Medical Center 12021 S. Wilmington Ave., Los Angeles, CA 90059.
s One US (NIH) unit is comparable to 1 International Unit of the First National Standard. 4 One KIU is equivalent to 1.111 X 10m3 trypsin inhibitor units. ’ One unit is that amount of Factor XIII which is contained in 1 ml of titrated, pooled human plasma. 175
oom4804/91
All rights
$1.50
0 1991 by Academic Press, Inc. of reproduction in any form reserved.
Copyright
176
JOURNAL 50
1
40.
RESEARCH:
VOL.
50,
NO.
2, FEBRUARY
1991
[
1: l .. *....
0 20. w
.
..
‘C,
IO-
ii O (L 0 50.
TEICOPLANIN
:40m ; JO=
SURGICAL
GENTAMICIN
CEFOXITIN
‘ii 30f.
: w
OF
FIBRIN
GLUE-CIPROFLOXACI
20.
5J
IO-
0
2
4
24 TIME (hr)
o-
1234567 TIME
FIG. 1. Antibiotic released from human sera (solid lines) and normal more than two-thirds of the antibiotic
(days)
fibrin glue-antibiotic clots into saline (dotted lines). Note that diffused out within 2 to 3 days.
polystyrene test tubes, followed by elution with pooled human serum (HS); separate elution studies were also performed using normal saline (NS). The test tubes were kept incubated at 37°C for 10 days during which time the supernatants were removed daily and replaced with fresh HS or NS. The supernatants were assayed daily for active concentrations of each Ab using a microbiological agar well diffusion method. In addition, the active concentration of each Ab within the FG-Ab clots was assayed following the addition of 0.5 ml of trypsin (to dissolve the clot) to a separate group of FG-Ab specimens. The antibacterial effects of FG-Ciprofloxacin and FG-Teicoplanin clots were evaluated by inoculating 1.0 ml of brain-heart infusion (BHI) with S. aureus (adjusted to give a concentration of lo5 colony forming units/ml), followed by the addition of 1.0 ml FG-Ab clots; a separate set of studies was also performed with the addition of 1.0 ml of mouse lung homogenate (MLH) as well. MLH was added because it is readily obtainable by standard techniques and dependably results in increased bacterial growth. The cultures were incubated at 37°C and aliquots drawn at 0,2,4, and 24 hr followed by serial dilution and plating on mannitol salt agar; the plates were incubated at 37°C for 24 hr and viable colony counts obtained.
FIG. 2. Growth of S. aure~ in brain-heart infusion alone and in the presence of fibrin glue and fibrin glue-Ciprofloxacin clots. Note the reduced bacterial growth in the presence of fibrin glue clots, which was even more pronounced in the presence of fibrin glue-Ciprofloxatin clots. Values shown are means + SEM.
which Ab release from FG-Ab clots was shown to be the result of simple diffusion through cross-linked FG clot interstices, which are greater than 1 pm in diameter [8]. Over 95% of the Ab could be recovered following lysis of the FG-Ab clots with trypsin, except in the case of FGGentamicin clots, in which only 85% could be recovered. The antibacterial effects of FG-Ciprofloxacin and FG-Teicoplanin clots in vitro were significant compared to those of FG clots not containing Ab (Figs. 2 and 3). The addition of either Ciprofloxacin or Teicoplanin to FG clots resulted in a reduction in bacterial growth of greater than lo4 CFU/ml of 5’. aureus at 24 hr compared to FG clots not containing Abs. In addition, the presence of FG alone (in BHI) resulted in a reduction in bacterial
13 12 II !
/
IO 5-
T
1
T
/
RESULTS 4J
The addition of Ab to FG clots resulted in continuous diffusion of the Ab into surrounding HS or NS for up to 5 to 7 days (Fig. 1). More than two-thirds of the Ab diffused out within 2 to 3 days, regardless of the Ab used or whether elution was performed with HS or NS. These results agree with those obtained in a previous study in
1 0
2
4
/m4 TIME (hr)
FIG. 3. Growth of S. aureu~ in brain-heart infusion alone and in the presence of fibrin glue and fibrin glue-Teicoplanin clots. Note the reduced bacterial growth in the presence of fibrin glue clots, which was even more pronounced in the presence of fibrin glue-Teicoplanin clots. Values shown are means k SEM.
KRAM
ET
AL.:
FIBRIN
GLUE-ANTIBIOTIC
177
MIXTURES
growth of between lo3 and lo4 CFU/ml at 24 hr compared to that of BHI alone. Thus, FG-Ab clots decreased bacterial growth at 24 hr by lo8 CFU/ml compared to BHI alone. The addition of MLH to BHI containing FG clots resulted in increased bacterial growth compared to that of FG and BHI alone. The addition of either Ciprofloxacin or Teicoplanin to the FG clots, however, inhibited the increase in bacterial growth secondary to the presence of MLH (Figs. 4 and 5). DISCUSSION
A potential disadvantage of all hemostatic agents is that they often must be left in surgical wounds after hemostasis has been achieved, resulting in the presence of a foreign body. Hinman and Babcock [9] have shown that a bacterial inoculum that is insufficient to produce infection may result in infection in the presence of a foreign body. Thus, although Dineen [lo] reported that oxidized cellulose may be relatively resistant to infection, Scher and Coil [ 111 have shown that both oxidized cellulose and microfibrillar collagen result in a greater frequency of infection compared to wounds not containing these agents. Data of the present study, on the other hand, revealed decreased growth of S. aureus (in BHI) in the presence of nonautologous FG compared to cultures not containing FG. This unexpected finding requires in vivo confirmation before it can be regarded as clinically important. However, clinical experience with nonautologous FG in over 150 potentially contaminated abdominal operations has not revealed an increased frequency of infec-
5J 0
2
4
f
TIME (hr1
24
FIG. 5. Growth of S. aureus in brain-heart infusion containing fibrin glue clots, fibrin glue clots and mouse lung homogenate (MLH), and fibrin glue-Teicoplanin clots and mouse lung homogenate. Note that although the addition of mouse lung homogenate to fibrin glue clots increased bacterial growth, this effect was inhibited by the presence of fibrin glue-Teicoplanin clots. Values shown are means -c SEM.
tion [12-141. This may be partially due to rapid degradation of the adhesive by fibrinolysis and complete removal by macrophages within 2 to 4 weeks, thereby eliminating any nidus for infection or chronic inflammatory reaction [2-6, 81. The addition of Ab to FG clots resulted in continuous diffusion of the Ab into surrounding HS or NS for up to 7 days; however, more than two-thirds of the Ab diffused out within 2 to 3 days. A method for decreasing Ab diffusion rates would be beneficial. Nevertheless, the presence of FG-Ab clots decreased bacterial growth (in BHI) by greater than 10’ CFU/ml at 24 hr compared to BHI alone. The addition of MLH to cultures containing S. aureus resulted in increased bacterial growth which was inhibited by the presence of FG-Ab clots. In conclusion, a variety of Abs can be combined with FG resulting in antibacterial concentrations for up to 5 to 7 days in vitro and FG alone inhibits S. aureus growth in vitro. Potential clinical applications of FG-Ab mixtures include sealing of gastrointestinal, vascular, and tracheobronchial anastomoses, and the achievement of hemostasis in potentially contaminated or infected wounds. REFERENCES
5’, 0
2
4
24 TIME (hr)
FIG. 4. Growth of S. aureu.s in brain-heart infusion containing fibrin glue clots, fibrin glue clots and mouse lung homogenate (MLH), and fibrin glue-Ciprofloxacin clots and mouse lung homogenate. Note that although the addition of mouse lung homogenate to fibrin glue clots increased bacterial growth, this effect was inhibited by the presence of fibrin glue-Ciprofloxacin clots. Values shown are means f SEM.
1.
2. 3.
Redl, H., Schlag, G., Dinges, H., et al. Background and methods of “fibrin sealing.” In G. D. Winter, D. F. Gibbons, and H. Plenk (Eds.), Biomateriak. New York: Wiley, 1982. Pp. 669-696. Kram, H. B., Shoemaker, W. C., Hino, S. T., et al. Splenic salvage using biologic glue. Arch. Surg. 119: 1309, 1984. Kram, H. B., Shoemaker, W. C., Hino, S. T., et al. Tracheal repair with fibrin glue. J. Thorac. Cardiovasc. Surg. 90: 771, 1985.
178
JOURNAL
OF
SURGICAL
RESEARCH:
4.
Jakob, H., Campbell, C. D., Qiu, Z. K., et al. Use of fibrin sealant for reinforcing arterial anastomoses. J. Vast. Surg. 1: 171, 1984.
5.
Kram, H. B., Garces, M. A., Klein, S. R., et al. Common bile duct anastomosis using fibrin glue. Arch. Surg. 120: 1250, 1985.
6.
Kram, H. B., Nugent, P., Reuben, B. I., et al. Fibrin glue sealing of polytetrafluoroethylene vascular graft anastomoses: Comparison with oxidized cellulose. J. Vast. Surg. 8: 563, 1988.
7.
Kram, H. B., Reuben, glue in hepatic trauma.
8.
Redl, tures
9.
Hinman,
B. I., Fleming, A. W., et al. Use J. Trauma 28: 1195, 1988.
H., Schlag, G., Stanek, G., et al. In vitro of fibrin seal and antibiotics. Biomaterials F., and Babcock,
K. 0. Local
reaction
10. 11.
cellu-
50,
NO.
2, FEBRUARY
1991
lose and gelatin hemostatic agents in experimentally contaminated renal wounds. Surgery 26: 633, 1949. Dineen, P. Antibacterial activity of oxidized regenerated cellulose. Surg. Gynecol. Obstet. 142: 481, 1976. Scher, K. microfibrillar
S., and Coil, J. A. Effects of oxidized collagen on infection. Surgery 91:
H. B., Nathan, R. C., Stafford, F., hemostasis in patients with coagulation
cellulose
and
301, 1982.
12.
Kram, achieves Surg.
13.
Kram, splenic
14.
Kram, H. B., Nathan, R. C., and Shoemaker, W. C. Fibrin renal and ureteral trauma. Urology 33: 215, 1989.
of fibrin
properties of mix4: 29, 1983. to oxidized
VOL.
et al. Fibrin disorders.
glue Arch.
124:385,1989. H. B., de1 Junco, T., Ocampo, H., et al. Techniques of preservation using fibrin glue. J. Trauma 30: 97, 1990. glue in
OF SURGICAL
RESEARCH
50,175-178
Antibacterial
(1991)
Effects of Fibrin Glue-Antibiotic
HARRY B. KRAM, M.D.,2 MAHDUBANSAL, Departments
of Surgery and Infectious and the UCLA School Submitted
PH.D.,
Mixtures’
OMATIMBERLAKE,ANDWILLIAMC.SHOEMAKER,
Disease, Los Angeles County King/Drew Medical of Medicine, Los Angeles, California 90024 for publication
February
M.D.
Center,
20, 1990
hemostasis and sealing anastomoses against leakage [l71. The adhesive is slowly dissolved over time by the body’s fibrinolytic system and is completely removed by macrophages within 2 to 4 weeks; normal wound healing progresses during this time. The combination of FG and antibiotics (Abs) has potential clinical applications in the sealing of gastrointestinal, vascular, and tracheobronchial anastomoses, as well as in the establishment of hemostasis in potentially contaminated or infected wounds. Ciprofloxacin and Teicoplanin are two newly developed Abs which are highly effective against a broad spectrum of bacteria. In the present in vitro study, we investigated the duration of action and antibacterial effects of various combined FG-Ab mixtures including Ciprofloxacin, Teicoplanin, Cefoxitin, and Gentamicin. In addition, the effect of FG alone on bacterial growth was evaluated.
In the present in vitro study, we investigated the duration of action and antibacterial effects of nonautologous fibrin glue (FG) combined with antibiotics (Abs) including Ciprofloxacin, Teicoplanin, Cefoxitin, and Gentamicin; the effect of FG alone on bacterial growth was also evaluated. The rate of Ab diffusion from combined FG-Ab clots was evaluated by separate elution with pooled human serum (HS) and normal saline (NS); supernatants were removed daily and assayed for active concentrations of each Ab. The effects of FG and combined FG-Ab clots on bacterial growth were evaluated by inoculating brain-heart infusion (BHI) with Staphylococcus aureus, followed by the addition of FG or FG-Ab clots; a separate set of studies was also performed with the addition of mouse lung homogenate (MLH) as well. The addition of Ab to FG clots resulted in continuous diffusion of the Ab into the surrounding HS or NS for up to 5 to 7 days; however, more than two-thirds of the Ab diffused out within 2 to 3 days regardless of the Ab used. The antibacterial effects of FG-Ciprofloxacin and FG-Teicoplanin clots were significant compared to those of FG clots not containing Ab. In addition, the presence of FG clots (in BHI) resulted in a reduction in bacterial growth compared to that of BHI alone. The addition of MLH to BHI resulted in increased bacterial growth, but this effect was inhibited by the presence of combined FG-Ab clots. In conclusion: (1) a variety of Abs can be combined with FG resulting in significant antibacterial effects in vitro, and (2) the presence of nonautologous FG alone inhibits S. aureus growth in vitro. Potential applications of FGAb mixtures include sealing of gastrointestinal, vascular, and tracheobronchial anastomoses, and the establishment of hemostasis in potentially contaminated or infected wounds. 0 1991 Academic Press, Inc.
MATERIALS Fibrin
AND METHODS
Glue
The FG (Immuno AG; Vienna, Austria) used in the study consisted of highly concentrated human fibrinogen, thrombin (500 US [NIH] units3/ml), calcium chloride (40 pm/ml), and aprotinin (3000 KIU*/ml). After preheating to 37”C, the fibrinogen was dissolved in aprotinin and the dried thrombin reconstituted in calcium chloride, with or without Ab. The components were drawn up into separate syringes which were connected to a single port that allowed simultaneous dispension and mixing of the components. The reconstituted adhesive contained 70 to 100 mg/ml of fibrinogen, 2 to 7 mg/ml of fibronectin (Cold Insoluble Globulin), 10 units5/ml of Factor XIII, and 35 pg/ml of plasminogen. Experimental
Protocol
Fibrin glue (FG), made with highly concentrated human fibrinogen and clotting factors, is a biologic adhesive that has been found to be effective in establishing
One milliliter FG-Ab clots containing 20 pg/ml of Ab were prepared and allowed to solidify at the bottom of
i Presented at the 15th Annual Meeting of the Society for Biomaterials, Lake Buena Vista, FL, April 28-May 2, 1989. ’ To whom reprint requests should be addressed at Department of Surgery, LA County King/Drew Medical Center 12021 S. Wilmington Ave., Los Angeles, CA 90059.
s One US (NIH) unit is comparable to 1 International Unit of the First National Standard. 4 One KIU is equivalent to 1.111 X 10m3 trypsin inhibitor units. ’ One unit is that amount of Factor XIII which is contained in 1 ml of titrated, pooled human plasma. 175
oom4804/91
All rights
$1.50
0 1991 by Academic Press, Inc. of reproduction in any form reserved.
Copyright
176
JOURNAL 50
1
40.
RESEARCH:
VOL.
50,
NO.
2, FEBRUARY
1991
[
1: l .. *....
0 20. w
.
..
‘C,
IO-
ii O (L 0 50.
TEICOPLANIN
:40m ; JO=
SURGICAL
GENTAMICIN
CEFOXITIN
‘ii 30f.
: w
OF
FIBRIN
GLUE-CIPROFLOXACI
20.
5J
IO-
0
2
4
24 TIME (hr)
o-
1234567 TIME
FIG. 1. Antibiotic released from human sera (solid lines) and normal more than two-thirds of the antibiotic
(days)
fibrin glue-antibiotic clots into saline (dotted lines). Note that diffused out within 2 to 3 days.
polystyrene test tubes, followed by elution with pooled human serum (HS); separate elution studies were also performed using normal saline (NS). The test tubes were kept incubated at 37°C for 10 days during which time the supernatants were removed daily and replaced with fresh HS or NS. The supernatants were assayed daily for active concentrations of each Ab using a microbiological agar well diffusion method. In addition, the active concentration of each Ab within the FG-Ab clots was assayed following the addition of 0.5 ml of trypsin (to dissolve the clot) to a separate group of FG-Ab specimens. The antibacterial effects of FG-Ciprofloxacin and FG-Teicoplanin clots were evaluated by inoculating 1.0 ml of brain-heart infusion (BHI) with S. aureus (adjusted to give a concentration of lo5 colony forming units/ml), followed by the addition of 1.0 ml FG-Ab clots; a separate set of studies was also performed with the addition of 1.0 ml of mouse lung homogenate (MLH) as well. MLH was added because it is readily obtainable by standard techniques and dependably results in increased bacterial growth. The cultures were incubated at 37°C and aliquots drawn at 0,2,4, and 24 hr followed by serial dilution and plating on mannitol salt agar; the plates were incubated at 37°C for 24 hr and viable colony counts obtained.
FIG. 2. Growth of S. aure~ in brain-heart infusion alone and in the presence of fibrin glue and fibrin glue-Ciprofloxacin clots. Note the reduced bacterial growth in the presence of fibrin glue clots, which was even more pronounced in the presence of fibrin glue-Ciprofloxatin clots. Values shown are means + SEM.
which Ab release from FG-Ab clots was shown to be the result of simple diffusion through cross-linked FG clot interstices, which are greater than 1 pm in diameter [8]. Over 95% of the Ab could be recovered following lysis of the FG-Ab clots with trypsin, except in the case of FGGentamicin clots, in which only 85% could be recovered. The antibacterial effects of FG-Ciprofloxacin and FG-Teicoplanin clots in vitro were significant compared to those of FG clots not containing Ab (Figs. 2 and 3). The addition of either Ciprofloxacin or Teicoplanin to FG clots resulted in a reduction in bacterial growth of greater than lo4 CFU/ml of 5’. aureus at 24 hr compared to FG clots not containing Abs. In addition, the presence of FG alone (in BHI) resulted in a reduction in bacterial
13 12 II !
/
IO 5-
T
1
T
/
RESULTS 4J
The addition of Ab to FG clots resulted in continuous diffusion of the Ab into surrounding HS or NS for up to 5 to 7 days (Fig. 1). More than two-thirds of the Ab diffused out within 2 to 3 days, regardless of the Ab used or whether elution was performed with HS or NS. These results agree with those obtained in a previous study in
1 0
2
4
/m4 TIME (hr)
FIG. 3. Growth of S. aureu~ in brain-heart infusion alone and in the presence of fibrin glue and fibrin glue-Teicoplanin clots. Note the reduced bacterial growth in the presence of fibrin glue clots, which was even more pronounced in the presence of fibrin glue-Teicoplanin clots. Values shown are means k SEM.
KRAM
ET
AL.:
FIBRIN
GLUE-ANTIBIOTIC
177
MIXTURES
growth of between lo3 and lo4 CFU/ml at 24 hr compared to that of BHI alone. Thus, FG-Ab clots decreased bacterial growth at 24 hr by lo8 CFU/ml compared to BHI alone. The addition of MLH to BHI containing FG clots resulted in increased bacterial growth compared to that of FG and BHI alone. The addition of either Ciprofloxacin or Teicoplanin to the FG clots, however, inhibited the increase in bacterial growth secondary to the presence of MLH (Figs. 4 and 5). DISCUSSION
A potential disadvantage of all hemostatic agents is that they often must be left in surgical wounds after hemostasis has been achieved, resulting in the presence of a foreign body. Hinman and Babcock [9] have shown that a bacterial inoculum that is insufficient to produce infection may result in infection in the presence of a foreign body. Thus, although Dineen [lo] reported that oxidized cellulose may be relatively resistant to infection, Scher and Coil [ 111 have shown that both oxidized cellulose and microfibrillar collagen result in a greater frequency of infection compared to wounds not containing these agents. Data of the present study, on the other hand, revealed decreased growth of S. aureus (in BHI) in the presence of nonautologous FG compared to cultures not containing FG. This unexpected finding requires in vivo confirmation before it can be regarded as clinically important. However, clinical experience with nonautologous FG in over 150 potentially contaminated abdominal operations has not revealed an increased frequency of infec-
5J 0
2
4
f
TIME (hr1
24
FIG. 5. Growth of S. aureus in brain-heart infusion containing fibrin glue clots, fibrin glue clots and mouse lung homogenate (MLH), and fibrin glue-Teicoplanin clots and mouse lung homogenate. Note that although the addition of mouse lung homogenate to fibrin glue clots increased bacterial growth, this effect was inhibited by the presence of fibrin glue-Teicoplanin clots. Values shown are means -c SEM.
tion [12-141. This may be partially due to rapid degradation of the adhesive by fibrinolysis and complete removal by macrophages within 2 to 4 weeks, thereby eliminating any nidus for infection or chronic inflammatory reaction [2-6, 81. The addition of Ab to FG clots resulted in continuous diffusion of the Ab into surrounding HS or NS for up to 7 days; however, more than two-thirds of the Ab diffused out within 2 to 3 days. A method for decreasing Ab diffusion rates would be beneficial. Nevertheless, the presence of FG-Ab clots decreased bacterial growth (in BHI) by greater than 10’ CFU/ml at 24 hr compared to BHI alone. The addition of MLH to cultures containing S. aureus resulted in increased bacterial growth which was inhibited by the presence of FG-Ab clots. In conclusion, a variety of Abs can be combined with FG resulting in antibacterial concentrations for up to 5 to 7 days in vitro and FG alone inhibits S. aureus growth in vitro. Potential clinical applications of FG-Ab mixtures include sealing of gastrointestinal, vascular, and tracheobronchial anastomoses, and the achievement of hemostasis in potentially contaminated or infected wounds. REFERENCES
5’, 0
2
4
24 TIME (hr)
FIG. 4. Growth of S. aureu.s in brain-heart infusion containing fibrin glue clots, fibrin glue clots and mouse lung homogenate (MLH), and fibrin glue-Ciprofloxacin clots and mouse lung homogenate. Note that although the addition of mouse lung homogenate to fibrin glue clots increased bacterial growth, this effect was inhibited by the presence of fibrin glue-Ciprofloxacin clots. Values shown are means f SEM.
1.
2. 3.
Redl, H., Schlag, G., Dinges, H., et al. Background and methods of “fibrin sealing.” In G. D. Winter, D. F. Gibbons, and H. Plenk (Eds.), Biomateriak. New York: Wiley, 1982. Pp. 669-696. Kram, H. B., Shoemaker, W. C., Hino, S. T., et al. Splenic salvage using biologic glue. Arch. Surg. 119: 1309, 1984. Kram, H. B., Shoemaker, W. C., Hino, S. T., et al. Tracheal repair with fibrin glue. J. Thorac. Cardiovasc. Surg. 90: 771, 1985.
178
JOURNAL
OF
SURGICAL
RESEARCH:
4.
Jakob, H., Campbell, C. D., Qiu, Z. K., et al. Use of fibrin sealant for reinforcing arterial anastomoses. J. Vast. Surg. 1: 171, 1984.
5.
Kram, H. B., Garces, M. A., Klein, S. R., et al. Common bile duct anastomosis using fibrin glue. Arch. Surg. 120: 1250, 1985.
6.
Kram, H. B., Nugent, P., Reuben, B. I., et al. Fibrin glue sealing of polytetrafluoroethylene vascular graft anastomoses: Comparison with oxidized cellulose. J. Vast. Surg. 8: 563, 1988.
7.
Kram, H. B., Reuben, glue in hepatic trauma.
8.
Redl, tures
9.
Hinman,
B. I., Fleming, A. W., et al. Use J. Trauma 28: 1195, 1988.
H., Schlag, G., Stanek, G., et al. In vitro of fibrin seal and antibiotics. Biomaterials F., and Babcock,
K. 0. Local
reaction
10. 11.
cellu-
50,
NO.
2, FEBRUARY
1991
lose and gelatin hemostatic agents in experimentally contaminated renal wounds. Surgery 26: 633, 1949. Dineen, P. Antibacterial activity of oxidized regenerated cellulose. Surg. Gynecol. Obstet. 142: 481, 1976. Scher, K. microfibrillar
S., and Coil, J. A. Effects of oxidized collagen on infection. Surgery 91:
H. B., Nathan, R. C., Stafford, F., hemostasis in patients with coagulation
cellulose
and
301, 1982.
12.
Kram, achieves Surg.
13.
Kram, splenic
14.
Kram, H. B., Nathan, R. C., and Shoemaker, W. C. Fibrin renal and ureteral trauma. Urology 33: 215, 1989.
of fibrin
properties of mix4: 29, 1983. to oxidized
VOL.
et al. Fibrin disorders.
glue Arch.
124:385,1989. H. B., de1 Junco, T., Ocampo, H., et al. Techniques of preservation using fibrin glue. J. Trauma 30: 97, 1990. glue in
