Diana S. Malcolm

Use of base deficit to compare resuscitation with lactated Ringer's solution, haemaccel, whole blood, and diaspirin cross-linked hemoglobin following hemorrhage in rats. Discussion

Base deficit (BD) has been shown to be a sensitive measure of the degree and duration of inadequate perfusion. We developed a rat model of hemorrhagic shock based on achieving a fixed BD of 13 +/- 1 mmol/L before resuscitation. Using this model, we compared the efficacy of resuscitation with lactated Ringers solution (LR), Haemaccel (a colloid), and whole blood with that of diaspirin cross-linked hemoglobin (DCLHb, Baxter Healthcare Corp.) by evaluating improvements in BD and restoration of base excess (BE, positive correlate of BD) for 60 minutes following resuscitation. The DCLHb was superior to LR and Haemaccel in restoring and maintaining BE following resuscitation, and was able to restore BE as rapidly as whole blood at half the volume. At 60 minutes, DCLHb at twice the shed blood volume maintained BE at higher (more positive) values compared with all other treatment groups. We conclude that DCLHb is at least as effective as whole blood and superior to LR and Haemaccel in restoring BE within the first 60 minutes following resuscitation in this hemorrhagic shock model.

Characterization of the Hemodynamic Response to Intravenous Diaspirin Crosslinked Hemoglobin Solution in Rats

Diaspirin crosslinked hemoglobin solution (DCLHB) has potential for clinical use as an oxygen-carrying solution because of its excellent oxygen transport properties and biochemical stability. The present study characterizes the effects of intravenous infusions of 0.625-40 mL/kg (62.5-4000 mg/kg) DCLHb on mean blood pressure (MAP) and heart rate (HR) in conscious rats. DCLHb at all doses tested except 62.5 mg/kg was associated with an immediate increase in MAP (25-30% above baseline) that peaked between 20-30 minutes after infusion and returned to baseline within 120-300 minutes in a dose-dependent manner. Maximum MAP achieved was in the range of 129 +/- 7 to 140 +/- 7 mm Hg and there was no statistically significant difference in the response between doses. HR responded in a reciprocal manner to changes in MAP. Volume- and oncotic-matched infusions of LR and albumin did not alter MAP or HR. Slow infusion (0.34 mL/min) of DCLHb appeared to blunt the magnitude of the pressor response when compared to bolus injection (< 10 sec). DCLHb administration is associated with a pressor response that is not due to volume load, oncotic pressure, or rate of infusion, suggesting that it is intrinsic to the modified hemoglobin molecule and pharmacologic in nature.

The efficacy of diaspirin crosslinked hemoglobin solution resuscitation in a model of uncontrolled hemorrhage.

Controversy exists whether early aggressive fluid therapy in the setting of uncontrolled hemorrhage worsens outcome by increasing blood loss from injured vessels. Since diaspirin crosslinked hemoglobin (DCLHb) is a vasoactive, oxygen-carrying solution, we compared the effects of DCLHb with other resuscitative fluids on blood loss, hemodynamics, and tissue oxygen delivery in a model of uncontrolled hemorrhage. Anesthetized rats (250-350 g) were subjected to a 50% tail transection and resuscitated 15 minutes later with 1:1 DCLHb, 3:1 lactated Ringers solution (LR), 1:1 hypertonic saline (7.5% HTS), or 1:1 human serum albumin (8.3% HSA) based on initial volume of blood loss (average 4.7 +/- 0.3 mL/kg). An unresuscitated group served as a control. Cumulative blood loss was measured at 5 hours postresuscitation. By 15 minutes after tail transection, mean arterial pressure (MAP) decreased 19.2 +/- 3.8 mm Hg from the baseline value (102 +/- 5 mm Hg). The DCLHb solution restored and maintained MAP and subcutaneous tissue oxygen tension at baseline values better than all other resuscitative fluids. Although blood loss in DCLHb-treated animals was greater than in unresuscitated animals, it was no different from other resuscitative fluids and less than with HSA. There was no difference in 24-hour survival between all treatment groups. In conclusion, DCLHb elevates MAP but does not exacerbate blood loss or compromise tissue oxygen delivery compared with other resuscitative fluids in this model of uncontrolled hemorrhage.

Subcutaneous oxygen tension : a useful adjunct in assessment of perfusion status

OBJECTIVES Using a new fluorescence-quenching optode which, unlike earlier oximeters, neither consumes oxygen nor generates heat, we sought to determine the effects of hemorrhage and resuscitation on subcutaneous PO2. Additionally, we compared the effects of resuscitation with diaspirin crosslinked hemoglobin, an oxygen-carrying solution, on subcutaneous PO2 to that of traditional resuscitative fluids. We also compared mean arterial pressure and central venous oxygen saturation, indirect indices of perfusion, to subcutaneous PO2, a direct index of perfusion. DESIGN Prospective trial, randomized for selection of treatment regimen. SETTING Shock-trauma laboratory of a medical university. SUBJECTS Male Sprague-Dawley rats, weighing 260 to 380 g. INTERVENTIONS Rats were bled 22 mL/kg and resuscitated, 1 min later, with either 66 mL/kg of lactated Ringers solution, 22 mL/kg of human serum albumin, 22 mL/kg of blood, or 22 mL/kg of diaspirin crosslinked hemoglobin. A fifth group of animals was not resuscitated after hemorrhage. Subcutaneous PO2 and mean arterial pressure were monitored continuously throughout the experiment, while central venous oxygen saturation was measured intermittently. MEASUREMENTS AND MAIN RESULTS Subcutaneous PO2 decreased in response to hemorrhage and, although it did increase after resuscitation with each fluid, no treatment was able to restore subcutaneous PO2 to baseline within 2 hrs postresuscitation. Subcutaneous PO2 continued to decrease after hemorrhage in the unresuscitated animals. In contrast, mean arterial pressure was restored to baseline values in only blood- and diaspirin crosslinked hemoglobin-treated animals, although this effect was lost within 30 mins in the blood-treated group. Only blood restored the central venous oxygen saturation to baseline values in the early postresuscitation period. CONCLUSIONS The fluorescence-quenching optode consistently followed changes in subcutaneous PO2 during hemorrhage and after resuscitation. Diaspirin crosslinked hemoglobin performed as well as blood in restoring peripheral perfusion, as measured by subcutaneous PO2, while both of these fluids were superior to either lactated Ringers solution or albumin. Both whole blood and diaspirin crosslinked hemoglobin restored mean arterial pressure to baseline, although the effect of the latter was of a longer duration. The pressor effect of the crosslinked hemoglobin did not affect peripheral perfusion, as reflected by the values for subcutaneous PO2. Subcutaneous PO2 is a useful adjunct in assessment of the adequacy of peripheral perfusion and may help redefine targets for resuscitation.

Diaspirin cross-linked hemoglobin is efficacious in gut resuscitation as measured by a GI tract optode.

The objective of this study was to compare the efficacy of diaspirin cross-linked hemoglobin (DCLHb) with that of standard resuscitative fluids in restoring intestinal mucosal oxygenation and villous architecture after hemorrhage. Male rats were bled to a base deficit of 5 +/- 2 nmol/l under propofol anesthesia and monitored for 90 minutes postresuscitation with DCLHb, blood, lactated Ringers solution, albumin, or nothing (DNR) for mucosal oxygen tension (Pmo2) and physiologic and laboratory parameters. Small intestinal histologic specimens were obtained and scored independently by two investigators blinded to therapy on a scale of 0 (normal) to 4 (worst). All treatments restored Pmo2; only DCLHb did so without exceeding baseline values. For untreated rats (DNR), Pmo2 was not restored. Normal mucosal architecture was maintained only in DCLHb-treated rats. As Pmo2 increased, mucosal score improved. In a rat model of controlled hemorrhage, Pmo2 changes measured by an optode correlated with gut histological abnormalities. By these criteria, DCLHb is superior to crystalloid, colloid, and blood in gut resuscitation.

Diaspirin Cross-Linked Hemoglobin Solution as a Resuscitatwe Fluid Following Severe Hemorrhage in the Rat

The effect of diaspirin cross-linked hemoglobin (DCLHb) on mean arterial pressure (MAP) and heart rate (HR) was compared to Ringers 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 maintained for at least 5 hrs. Hemodynamic responses in DCLHb-treated animals were not significantly different from 35 cc/kg blood-treated animals. In RL (105 cc/kg) treated animals the MAP was restored to 60-70% of baseline. 24 hr survival in animals resuscitated with fluids ranged between 88-100% and was not significantly different between treatment groups.

Early resuscitation with low-volume polyDCLHb is effective in the treatment of shock induced by penetrating vascular injury

OBJECTIVE To study the efficacy of an oxygen-carrying solution in early resuscitation of hemorrhagic shock induced by penetrating vascular injury. DESIGN Experimental study with anesthetized rats. MATERIALS AND METHODS Severe hemorrhagic shock was induced by a 25-gauge needle puncture to the infrarenal aorta. Forty animals were resuscitated 10 minutes after injury with either lactated Ringers solution (LR; 60 mL/kg), 7.5% hypertonic saline (HTS; 5 mL/kg), or modified diaspirin cross-linked hemoglobin (PolyDCLHb; 5 or 20 mL/kg) or were not resuscitated (NR) and followed for 6 hours. RESULTS Total blood loss was similar in all treatment groups. Mean arterial pressure was restored to baseline values, base deficit was corrected to base excess, and venous oxygen saturation improved with PolyDCLHb and more slowly with LR but persisted below baseline values with HTS and NR. The 6-hour mortality rates were zero of eight (low-dose PolyDCLHb), three of eight (high-dose PolyDCLHb), two of eight (LR), six of eight (HTS), and six of eight (NR). CONCLUSION Early resuscitation with low-volume hemoglobin is effective in restoring tissue perfusion and improving survival in uncontrolled hemorrhagic shock.

Diaspirin Crosslinked Hemoglobin (DCLHbTM): Control of Pressor Effect with Anti-Hypertensive Agents

Diaspirin crosslinked hemoglobin (DCLHb) administration elevates mean arterial pressure (MAP). The purpose of this study was to determine whether commonly used antihypertensive agents could control this pressor effect in rats. Awake rats were injected intravenously (i.v.) with 280 mg/kg of DCLHb. Fifteen minutes later when MAP was 25-30% above baseline and heart rate (HR) was reciprocally decreased, prazosin (2 mg/kg;an alpha adrenergic blocker), nitroglycerine (NTG; 10-150 mcg/min; a nitrovasodilator), nicardipine (0.204-0.08 mg/hr; a calcium channel blocker) or labetalol (5 mg/kg; an alpha/beta adrenergic blocker) was administered i.v. All four classes of antihypertensive agents promptly restored MAP to baseline. Coincident with the return of MAP to baseline, HR was restored to baseline in prazosin and NTG treated animals, however, bradycardia persisted in those animals treated with nicardipine and labetalol, most likely due to the negative chronotropic properties of these agents. We conclude that the pressor effect of DCLHb can be readily controlled with at least four different classes of commonly used antihypertensive agents.

Diaspirin Crosslinked Hemoglobin (DCLHb™) Attenuates Bacterial Translocation in Rats

Intestinal barrier function is compromised following severe hemorrhage which may allow bacterial translocation (BT) to occur and subsequently initiate a systemic response leading to multiple system organ failure (MSOF). This study compared BT following hemorrhage and resuscitation with lactated Ringers solution (LR) or diaspirin crosslinked hemoglobin solution (DCLHb). Rats (250-350 grams) were hemorrhaged to a base deficit of 15 +/- 2 mmol/L and immediately resuscitated with either 3:1 LR or 1:1 DCLHb based on shed blood volume. Four hours following resuscitation, the mesenteric lymph node complex was harvested, homogenized and plated onto MacConkey and Columbia CNA agar culture media. Facultative anaerobic and obligate aerobic bacteria were identified 48 hours later in 11/22 (50%) LR-treated rats and in 4/21 (19%) DCLHb-treated rats (p < or = 0.05). Following resuscitation, base excess (BE) and central venous oxygen saturation (SvO2) were not only restored to baseline but were significantly greater (p < or = 0.05) in DCLHb-treated rats than in LR-treated rats. In a separate group of rats subjected to the same hemorrhage and resuscitation protocol, mean arterial pressure in DCLHb-treated rats, but not LR-treated rats, was restored to baseline by 15 minutes and remained at or above baseline for up to 4 hrs. Twenty-four hour survival was 50% in LR-treated rats and 77% in DCLHb-treated rats (p > 0.05). These data suggest that DCLHb is superior to LR in restoring tissue oxygen delivery, as judged by BE and SvO2. Furthermore, since DCLHb restores oxygen delivery and attenuates BT, early resuscitation with DCLHb may limit gut ischemia and subsequent gut barrier failure and hence prevent the development of sepsis, MSOF and subsequent death.