Biomaterials, Artificial Cells and Immobilization Biotechnology
ISSN: 1055-7172 (Print) (Online) Journal homepage: http://www.tandfonline.com/loi/ianb18
Diaspirin Cross-Linked Hemoglobin Solution as a Resuscitatwe Fluid Following Severe Hemorrhage in the Rat Diana Malcolm, David Kissinger & Magnus Garrioch To cite this article: Diana Malcolm, David Kissinger & Magnus Garrioch (1992) Diaspirin CrossLinked Hemoglobin Solution as a Resuscitatwe Fluid Following Severe Hemorrhage in the Rat, Biomaterials, Artificial Cells and Immobilization Biotechnology, 20:2-4, 495-497, DOI: 10.3109/10731199209119674 To link to this article: http://dx.doi.org/10.3109/10731199209119674
Published online: 11 Jul 2009.
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Citing articles: 7 View citing articles
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Date: 18 April 2016, At: 21:10
B I O M T . , ART. CELLS & IPlWOB. BIOTECH., 2 0 ( 2 - 4 ) , 4 9 5 - 4 9 7 ( 1 9 9 2 )
Downloaded by [RMIT University Library] at 21:10 18 April 2016
DIASPIRlN CROSS-LINKED HEMOGLOBIN SOLUTION AS A RESUSCITATIVE FLUID FOLLOWING SEVERE HEMORRHAGE IN THE RAT
Diana Malcolm, Ph.D, David Kissinger, M.D., Magnus Garrioch, M.D. Uniformed Services University of the Health Sciences, Bethesda, MD Dept. of Shockflraurna, Washington Hospital Center, Washington, D.C. The Meet of dimpirin cross-linked hemoglobin (DCLHb) on mean Merial pressure (MAP) and heart rate (HR) wm compared to Ringer’s lactate (RL.)and shed blood in a 70% lethal model of hemorrhage (35 cc/kg blood loss) in conscious rats. All animals resuscitated with DCLHb regardless of dose (17.5 and 35 cc/kg) and concentration (7% and 10% solution) exhibited complete restoration of MAP and HR which was maintahed for at least 5 hrs. Hemodynamic responses in DCLHb-treated animals were not sign@cant& different from 35 cc/lsg blood-treated animals. In RL (105 cc/kg) treated animals the MAP was restored to 6070% of baseline. 24 hr survival in animals resuscitated with fluids ranged between 88-100% and was not sign@cant& different between treatment groups.
m u c t i o n ; Diaspirin cross-linked hemoglobin (DCLHb), a chemically stabilized stromafree hemoglobin solution, holds promise as a resuscitative fluid because of its excellent oxygen-transport properties and biochemical stability (1). In addition, these solutions do not require cross-matching or typing prior to administration, are less viscous than whole blood and may be better able to carry oxygen through narrowed vessels to ischemic tissues due to the smaller size of the hemoglobin molecule relative to the red cell. These solutions are also devoid of white blood cells and other blood componentswhich are known to contribute to ischemic tissue injury by releasing cytotoxic products. Finally, stroma-free hemoglobin solutions have oncotic properties which may be useful in recruiting interstitial fluids into the intravascular space to augment fluid resuscitation. The purpose of the present studies was to test the efficacy of DCLHb as a resuscitative fluid in a lethal model of hemorrhage in the conscious rat. The ability of DCLHb to restore and maintain mean arterial pressure and heart rate was compared to that of standard crystalloid therapy (3:l Ringer’s lactate) and whole blood. Twenty-four hour suMval was also compared between treatment groups. Male, Sprague-Dawley rats, weighing 275-350 g, were anesthetized with combination ketamine (60 mg/kg)/pentobarbital(22 mg/kg). The jugular vein and tail artery were catheterized for blood withdrawal/infusions and cardiovascular monitoring, respectively. Animals were allowed to recover from surgery and anesthesia for 24 hours prior to study. On the experimental day, conscious, unrestrained rats were bled 35 cc/kg at approximately 1 cc/min. Twenty minutes after the end of bleed, either shed blood (35 cc/kg), Ringer’s lactate (RL,105 cc/kg) or DCLHb (7% or 10%; 17.5 and 35 cc/kg) were infused at 1 cc/min except for the RL which was infused at 3 cc/min. A separate group of 495
Copyright 8 1992 by Mucel Dekker, Inc.
496
MALCOLM, KISSINGER, AND GARRIOCH
200 -. 180
-= 0
E
Downloaded by [RMIT University Library] at 21:10 18 April 2016
v
4
0-om-
--
o-owgso/lp) A- A L (106 -/lo)
160 --
A-A
0-0
140-
.-WX
1OrDQH(17.8& l a DQH (3s0/110) #xHb (17.8 co/lp)
120.-
10080
--
60-40 --
20t -
m a
min POI
(RL)and shed blood on mean arterial pressure (MAP) responses to hemorrhage (35 cc/kg) and resuscitation (n=8-9).
Figme 1. Effects of DCLHb, Ringer's Lactate
rats was not resuscitated (n= 15). Mean arterial pressure (MAP) and heart rate (HR) were monitored for up to 5 hours following infusion. 'Ibenty-four hour survival was monitored. The number of animals in each treatment group was 8-9. Prior to infusion, the DCLHb solutions were passed through a 0.2-0.45 M r n pore size cellulose acetate synnge filter. Methemoglobin concentration was determined spectrophotometricallyand ranged from 6510%. All test solutions were warmed to body temperature (37oC; before infusing. Resplts: The results are presented in Figure 1. Following the 35 cc/kg bleed, which is approximately one-half of the estimated blood volume, MAP dropped to 31 2 3 mm Hg from an average baseline of 99.9+4 mm Hg. Within 20 minutes (at the time of fluid resuscitation), MAP had returned to approximately 57% of baseline with the development of tachycardia. This initial increase in MAP is due to physiologic compensatory mechanisms including a baroreceptor reflex mediated increase in HR. The MAP of animals not resuscitated with fluids remained at 50-55 mm Hg throughout the observation period except shortly before death when MAP plummeted. At 24 hrs post-resuscitation, only 4 out of 15 unresuscitated animals were alive (73% mortality). In animals resuscitated with 3:l RL,MAP increased to 80% of baseline during the infusion, however, by the end of infusion, MAP fell slightly, probably reflecting redistribution of fluid volume. MAP remained at 60-70% of baseline throughout the 5 hr observation period. All &treated rats (n-8) survived 24 hrs, although they were significantly tachycardic (4%500 bpm) and had a wide pulse pressure (*SO mm Hg range).
Downloaded by [RMIT University Library] at 21:10 18 April 2016
R E S U S C I T A T I V E F L U I D FOR S E V E R E HEMORRHAGE
497
All animals resuscitated with DCLHb,regardless of dose (17.5or 35 cc/kg) and concentration (7%. lo%), exhibited similar hemodynamic responses to resuscitation compared to the animals receiving shed blood. Animals which received the 10% DCLHb (17.5 and 35 cc/kg) solution exhibited a higher MAP (12023 mm Hg) and lower HR (351221 bpm) than shed blood (103k6 mm Hg; 444220 bpm) at 60 minutes postresuscitation. At 120-300minutes post-resuscitation,MAP and HR were not significantly different between DLCHb and blood-treated groups. At 24 hrs the survival was: 7/8 for blood; 8/9 for 10% DCLHb (17.5 cc/kg); 7/8 for 10% DCLHb (35 cc/kg); 8/9 for 7% DCLHb (17.5 cc/kg) and 8/8 for 7% DCLHb (35 cc/kg). Discussioa: Diaspirin-cross linked hemoglobin solution holds promise as a resuscitative fluid. Findings from our hemorrhagic shock study suggest that 7% DCLHb is as efficacious as 10% DCLHb in restoring and maintaining MAP and HR. Similarly, DCLHb infused at half the volume of shed blood was as efficacious as shed blood in restoring cardiovascular function. This observation confirms our earlier study in anesthetized rats (2) in which a dose of DCLHb equal to half the volume of shed blood improved cardiovascular function and restored oxygen delivery to the skin as well as resuscitation with an equal volume of whole blood. The present study in conscious rats represents a more lethal model of hemorrhage (70%) than the previous study (2)and our findings that DCLHb is as effective as blood in restoring hemodynamics and in preserving survival further attest to the suitability of DCLHb as a resuscitative fluid. We also observed an increase over baseline in MAP following DCLHb infusion (all doses). This hemoglobin-induced hypertension has been reported by us (2) and by others (3,4)and is most likely due to the vasoactive properties of hemoglobin solutions (5,6). We have subsequentlyshown that the DCLHb-induced hypertension is mediated at least in part by an increase in systemic vascular resistance (data not reported). In our earlier study (2) we reported that concomitantwith an increase in MAP, tissue oxygenation was restored and maintained as well in DCLHb resuscitated animals as those resuscitated with blood. Thus, despite the increase in MAP and systemic resistance, DCLHb maintains tissue oxygen delivery which is paramount to successful shock treatment. In summary, DCLHb is as efficacious as whole blood and superior to Ringer's lactate in restoring hernodynamics in a lethal model of hemorrhagic shock. Clearly, hemoglobin solutions have the potential to become important adjunct resuscitative fluids. DCLHb may be useful in the initial resuscitation of severe hypovolemia that is refractory to standard crystalloid therapy and when oxygen-carrying capacity is critical. DCLHb provides immediate administration of an oxygen-carryingsolution to oxygen-deprived tissues and may ''buy'' the time before more definitive medical support (i.e., blood, surgery) is available.
2. 3. 4.
Snyder SR, Welty EV, Walder RY, Williams LA,Walder JA. Proc Natl Acad Sci ysB 84:7280, 1987. Przybelski RJ, Malcolm DS, Bums DG, Winslow RM. J J a b Clin Med 2:143,1991. Pharm The1 23:73, 1978. Savitsky JP, Doni J, Black J, Arnold JD. Jesch FH. Peters W,Hobbhahn J, Schoenberg M, Messmer K. Grit Care Med 10270,
5. 6.
Feola M, Simoni J, Fishman D, Tran R, Canizaro PC. 13(3):209, 1989. Martin W, Villani GM, Jothianandan D, Furchgott RF. j Pharmacol Fxp
1.
1982. 232:708,1985.
ISSN: 1055-7172 (Print) (Online) Journal homepage: http://www.tandfonline.com/loi/ianb18
Diaspirin Cross-Linked Hemoglobin Solution as a Resuscitatwe Fluid Following Severe Hemorrhage in the Rat Diana Malcolm, David Kissinger & Magnus Garrioch To cite this article: Diana Malcolm, David Kissinger & Magnus Garrioch (1992) Diaspirin CrossLinked Hemoglobin Solution as a Resuscitatwe Fluid Following Severe Hemorrhage in the Rat, Biomaterials, Artificial Cells and Immobilization Biotechnology, 20:2-4, 495-497, DOI: 10.3109/10731199209119674 To link to this article: http://dx.doi.org/10.3109/10731199209119674
Published online: 11 Jul 2009.
Submit your article to this journal
Article views: 1
View related articles
Citing articles: 7 View citing articles
Full Terms & Conditions of access and use can be found at http://www.tandfonline.com/action/journalInformation?journalCode=ianb18 Download by: [RMIT University Library]
Date: 18 April 2016, At: 21:10
B I O M T . , ART. CELLS & IPlWOB. BIOTECH., 2 0 ( 2 - 4 ) , 4 9 5 - 4 9 7 ( 1 9 9 2 )
Downloaded by [RMIT University Library] at 21:10 18 April 2016
DIASPIRlN CROSS-LINKED HEMOGLOBIN SOLUTION AS A RESUSCITATIVE FLUID FOLLOWING SEVERE HEMORRHAGE IN THE RAT
Diana Malcolm, Ph.D, David Kissinger, M.D., Magnus Garrioch, M.D. Uniformed Services University of the Health Sciences, Bethesda, MD Dept. of Shockflraurna, Washington Hospital Center, Washington, D.C. The Meet of dimpirin cross-linked hemoglobin (DCLHb) on mean Merial pressure (MAP) and heart rate (HR) wm compared to Ringer’s lactate (RL.)and shed blood in a 70% lethal model of hemorrhage (35 cc/kg blood loss) in conscious rats. All animals resuscitated with DCLHb regardless of dose (17.5 and 35 cc/kg) and concentration (7% and 10% solution) exhibited complete restoration of MAP and HR which was maintahed for at least 5 hrs. Hemodynamic responses in DCLHb-treated animals were not sign@cant& different from 35 cc/lsg blood-treated animals. In RL (105 cc/kg) treated animals the MAP was restored to 6070% of baseline. 24 hr survival in animals resuscitated with fluids ranged between 88-100% and was not sign@cant& different between treatment groups.
m u c t i o n ; Diaspirin cross-linked hemoglobin (DCLHb), a chemically stabilized stromafree hemoglobin solution, holds promise as a resuscitative fluid because of its excellent oxygen-transport properties and biochemical stability (1). In addition, these solutions do not require cross-matching or typing prior to administration, are less viscous than whole blood and may be better able to carry oxygen through narrowed vessels to ischemic tissues due to the smaller size of the hemoglobin molecule relative to the red cell. These solutions are also devoid of white blood cells and other blood componentswhich are known to contribute to ischemic tissue injury by releasing cytotoxic products. Finally, stroma-free hemoglobin solutions have oncotic properties which may be useful in recruiting interstitial fluids into the intravascular space to augment fluid resuscitation. The purpose of the present studies was to test the efficacy of DCLHb as a resuscitative fluid in a lethal model of hemorrhage in the conscious rat. The ability of DCLHb to restore and maintain mean arterial pressure and heart rate was compared to that of standard crystalloid therapy (3:l Ringer’s lactate) and whole blood. Twenty-four hour suMval was also compared between treatment groups. Male, Sprague-Dawley rats, weighing 275-350 g, were anesthetized with combination ketamine (60 mg/kg)/pentobarbital(22 mg/kg). The jugular vein and tail artery were catheterized for blood withdrawal/infusions and cardiovascular monitoring, respectively. Animals were allowed to recover from surgery and anesthesia for 24 hours prior to study. On the experimental day, conscious, unrestrained rats were bled 35 cc/kg at approximately 1 cc/min. Twenty minutes after the end of bleed, either shed blood (35 cc/kg), Ringer’s lactate (RL,105 cc/kg) or DCLHb (7% or 10%; 17.5 and 35 cc/kg) were infused at 1 cc/min except for the RL which was infused at 3 cc/min. A separate group of 495
Copyright 8 1992 by Mucel Dekker, Inc.
496
MALCOLM, KISSINGER, AND GARRIOCH
200 -. 180
-= 0
E
Downloaded by [RMIT University Library] at 21:10 18 April 2016
v
4
0-om-
--
o-owgso/lp) A- A L (106 -/lo)
160 --
A-A
0-0
140-
.-WX
1OrDQH(17.8& l a DQH (3s0/110) #xHb (17.8 co/lp)
120.-
10080
--
60-40 --
20t -
m a
min POI
(RL)and shed blood on mean arterial pressure (MAP) responses to hemorrhage (35 cc/kg) and resuscitation (n=8-9).
Figme 1. Effects of DCLHb, Ringer's Lactate
rats was not resuscitated (n= 15). Mean arterial pressure (MAP) and heart rate (HR) were monitored for up to 5 hours following infusion. 'Ibenty-four hour survival was monitored. The number of animals in each treatment group was 8-9. Prior to infusion, the DCLHb solutions were passed through a 0.2-0.45 M r n pore size cellulose acetate synnge filter. Methemoglobin concentration was determined spectrophotometricallyand ranged from 6510%. All test solutions were warmed to body temperature (37oC; before infusing. Resplts: The results are presented in Figure 1. Following the 35 cc/kg bleed, which is approximately one-half of the estimated blood volume, MAP dropped to 31 2 3 mm Hg from an average baseline of 99.9+4 mm Hg. Within 20 minutes (at the time of fluid resuscitation), MAP had returned to approximately 57% of baseline with the development of tachycardia. This initial increase in MAP is due to physiologic compensatory mechanisms including a baroreceptor reflex mediated increase in HR. The MAP of animals not resuscitated with fluids remained at 50-55 mm Hg throughout the observation period except shortly before death when MAP plummeted. At 24 hrs post-resuscitation, only 4 out of 15 unresuscitated animals were alive (73% mortality). In animals resuscitated with 3:l RL,MAP increased to 80% of baseline during the infusion, however, by the end of infusion, MAP fell slightly, probably reflecting redistribution of fluid volume. MAP remained at 60-70% of baseline throughout the 5 hr observation period. All &treated rats (n-8) survived 24 hrs, although they were significantly tachycardic (4%500 bpm) and had a wide pulse pressure (*SO mm Hg range).
Downloaded by [RMIT University Library] at 21:10 18 April 2016
R E S U S C I T A T I V E F L U I D FOR S E V E R E HEMORRHAGE
497
All animals resuscitated with DCLHb,regardless of dose (17.5or 35 cc/kg) and concentration (7%. lo%), exhibited similar hemodynamic responses to resuscitation compared to the animals receiving shed blood. Animals which received the 10% DCLHb (17.5 and 35 cc/kg) solution exhibited a higher MAP (12023 mm Hg) and lower HR (351221 bpm) than shed blood (103k6 mm Hg; 444220 bpm) at 60 minutes postresuscitation. At 120-300minutes post-resuscitation,MAP and HR were not significantly different between DLCHb and blood-treated groups. At 24 hrs the survival was: 7/8 for blood; 8/9 for 10% DCLHb (17.5 cc/kg); 7/8 for 10% DCLHb (35 cc/kg); 8/9 for 7% DCLHb (17.5 cc/kg) and 8/8 for 7% DCLHb (35 cc/kg). Discussioa: Diaspirin-cross linked hemoglobin solution holds promise as a resuscitative fluid. Findings from our hemorrhagic shock study suggest that 7% DCLHb is as efficacious as 10% DCLHb in restoring and maintaining MAP and HR. Similarly, DCLHb infused at half the volume of shed blood was as efficacious as shed blood in restoring cardiovascular function. This observation confirms our earlier study in anesthetized rats (2) in which a dose of DCLHb equal to half the volume of shed blood improved cardiovascular function and restored oxygen delivery to the skin as well as resuscitation with an equal volume of whole blood. The present study in conscious rats represents a more lethal model of hemorrhage (70%) than the previous study (2)and our findings that DCLHb is as effective as blood in restoring hemodynamics and in preserving survival further attest to the suitability of DCLHb as a resuscitative fluid. We also observed an increase over baseline in MAP following DCLHb infusion (all doses). This hemoglobin-induced hypertension has been reported by us (2) and by others (3,4)and is most likely due to the vasoactive properties of hemoglobin solutions (5,6). We have subsequentlyshown that the DCLHb-induced hypertension is mediated at least in part by an increase in systemic vascular resistance (data not reported). In our earlier study (2) we reported that concomitantwith an increase in MAP, tissue oxygenation was restored and maintained as well in DCLHb resuscitated animals as those resuscitated with blood. Thus, despite the increase in MAP and systemic resistance, DCLHb maintains tissue oxygen delivery which is paramount to successful shock treatment. In summary, DCLHb is as efficacious as whole blood and superior to Ringer's lactate in restoring hernodynamics in a lethal model of hemorrhagic shock. Clearly, hemoglobin solutions have the potential to become important adjunct resuscitative fluids. DCLHb may be useful in the initial resuscitation of severe hypovolemia that is refractory to standard crystalloid therapy and when oxygen-carrying capacity is critical. DCLHb provides immediate administration of an oxygen-carryingsolution to oxygen-deprived tissues and may ''buy'' the time before more definitive medical support (i.e., blood, surgery) is available.
2. 3. 4.
Snyder SR, Welty EV, Walder RY, Williams LA,Walder JA. Proc Natl Acad Sci ysB 84:7280, 1987. Przybelski RJ, Malcolm DS, Bums DG, Winslow RM. J J a b Clin Med 2:143,1991. Pharm The1 23:73, 1978. Savitsky JP, Doni J, Black J, Arnold JD. Jesch FH. Peters W,Hobbhahn J, Schoenberg M, Messmer K. Grit Care Med 10270,
5. 6.
Feola M, Simoni J, Fishman D, Tran R, Canizaro PC. 13(3):209, 1989. Martin W, Villani GM, Jothianandan D, Furchgott RF. j Pharmacol Fxp
1.
1982. 232:708,1985.
