Endovascular Embolization Techniques in a Novel Swine Model of Fatal Uncontrolled Solid Organ Hemorrhage and Coagulopathy.

BACKGROUND: Endovascular embolization is increasingly used in treating traumatic hemorrhage and other applications. No endovascular-capable translational large animal models exist and coagulopathy's effect on embolization techniques is unknown. We developed a coagulation-adaptable solid organ hemorrhage model in swine for investigation of embolization techniques. METHODS: Anesthetized swine (n = 26, 45 ± 3 kg) had laparotomy and splenic externalization. Half underwent 50% isovolemic hemodilution with 6% hetastarch and cooling to 33-35°C (COAG group). All had controlled 20 mL/kg hemorrhage and endovascular access to the proximal splenic artery with a 4F catheter via a right femoral sheath. Splenic transection and 5 min free bleeding were followed by treatment (n = 5/group) with 5 mL gelfoam slurry, three 6-mm coils, or no treatment (n = 3, control). Animals received 15 mL/kg plasma resuscitation and were monitored for 6 hr. Splenic blood loss was continuously measured and angiograms were performed at specified times. RESULTS: Coagulopathy was successfully established in COAG animals. Pre-treatment blood loss was greater in COAG (11 ± 6 mL/kg) than non-COAG (7 ± 3 mL/kg, P = 0.04) animals. Splenic hemorrhage was universally fatal without treatment. Non-COAG coil survival was 4/5 (326 ± 75 min) and non-COAG Gelfoam 3/5 (311 ± 67 min) versus non-COAG Control 0/3 (82 ± 18 min, P < 0.05 for both). Neither COAG Coil (0/5, 195 ± 117 min) nor COAG Gelfoam (0/5, 125 ± 32 min) treatment improved survival over COAG Control (0/3, 56 ± 19 min). Post-treatment blood loss was 4.6 ± 3.4 mL/kg in non-COAG Coil and 4.6 ± 2.9 mL/kg in non-COAG Gelfoam, both lower than non-COAG Control (18 ± 1.3 mL/kg, P = 0.05). Neither COAG Coil (8.4 ± 5.4 mL/kg) nor COAG Gelfoam (15 ± 11 ml/kg) had significantly less blood loss than COAG Control (20 ± 1.2 mL/kg). Both non-COAG treatment groups had minimal blood loss during observation, while COAG groups had ongoing slow blood loss. In the COAG Gelfoam group, there was an increase in hemorrhage between 30 and 60 min following treatment. CONCLUSIONS: A swine model of coagulation-adaptable fatal splenic hemorrhage suitable for endovascular treatment was developed. Coagulopathy had profound negative effects on coil and gelfoam efficacy in controlling bleeding, with implications for trauma and elective embolization procedures.

Differing Resuscitation With Aortic Occlusion in a Swine Junctional Hemorrhage Polytrauma Model.

BACKGROUND: Resuscitative endovascular balloon occlusion of the aorta (REBOA) and Abdominal Aortic and Junctional Tourniquet (AAJT) have received much attention in recent as methods for temporary control of junctional hemorrhage. Previous studies typically used the animal's shed blood for resuscitation. With current interest in moving REBOA to prehospital environment, this study aimed to evaluate the hemodynamic and metabolic responses to different resuscitation fluids used with these devices. METHODS: In swine (Sus scrofa), shock was induced using a controlled hemorrhage, femur fracture, and uncontrolled hemorrhage from the femoral artery. Infrarenal REBOA or AAJT was deployed for 60 min during which the arterial injury was repaired. Animals were resuscitated with 15 mL/kg of shed whole blood (SWB) or fresh frozen plasma (FFP) or 30 mL/kg of a balanced crystalloid (PlasmaLyte). RESULTS: Animals in the AAJT and REBOA groups did not show any measurable differences in hemodynamics, metabolic responses, or survival with AAJT or REBOA treatment; hence, the data are pooled and analyzed among the three resuscitative fluids. SWB, FFP, and PlasmaLyte groups did not have a difference in survival time or overall survival. The animals in the SWB and FFP groups maintained higher blood pressure after resuscitation, (P < 0.001) and required significantly less norepinephrine to maintain blood pressure than those in the PlasmaLyte group (P < 0.001). The PlasmaLyte resuscitation prolonged prothrombin time and decreased thromboelastography maximum amplitude. CONCLUSIONS: After 60 min, infrarenal REBOA or AAJT aortic occlusion SWB and FFP resuscitation provided better blood pressure support with half of the resuscitative volume of PlasmaLyte. Swine resuscitated with SWB and FFP also had a more favorable coagulation profile. These data suggest that whole blood or component therapy should be used for resuscitation in conjunction with REBOA or AAJT, and administration of these fluids should be considered if prehospital device use is pursued.

Long-term consequences of abdominal aortic and junctional tourniquet for hemorrhage control.

BACKGROUND: Specialized tourniquets have been deployed to the battlefield for the control of junctional/pelvic hemorrhage despite limited knowledge concerning their safety and duration of use. This study investigated long-term effects of abdominal application of the abdominal aortic and junctional tourniquet (AAJT) in a swine survival model. METHODS: Anesthetized spontaneously air-breathing swine were subjected to bilateral femoral artery injuries and subsequent 40% hemorrhage. Further hemorrhage was controlled by applying the AAJT on the lower abdomen for 0 h (n = 2, controls), 1 h (n = 6), 1.5 h (n = 6), or 2 h (n = 3). Before tourniquet release, arterial injuries were repaired, and mechanical ventilation and rapid crystalloid fluid were provided for at least 5 min. Additional fluid and 500 mL autologous blood were transfused after restoring blood flow. Animals were recovered and their mobility and health monitored up to 2 wk. RESULTS: AAJT application occluded the infrarenal abdominal aorta and stopped bilateral groin hemorrhage with rapid reversal of hemorrhagic shock and improved cranial blood pressure. All animals including controls recovered overnight but regaining hind leg function varied among AAJT-treated groups. In contrast to 1 h AAJT-treated swine that recovered full mobility in 1 wk, 2 h animals developed persistent hind leg paraplegia concurrent with urinary retention and ischemic necrosis of lumber muscles and had to be euthanized 3 d after surgery. Half of the 1.5-h group also had to be euthanized early due to paraplegia, whereas the other half recovered motor function within 2 wk. CONCLUSIONS: The results of this animal study indicated that ischemic reperfusion injuries associated with abdominal application of the AAJT were time-dependent. To avoid permanent injuries, AAJT application on the abdomen to control a groin hemorrhage could not be longer than 1 h. This was consistent with recent instructions for application of this tourniquet on the abdomen in patients.

Diagnostic accuracy of a novel software technology for detecting pneumothorax in a porcine model.

INTRODUCTION: Our objective was to measure the diagnostic accuracy of a novel software technology to detect pneumothorax on Brightness (B) mode and Motion (M) mode ultrasonography. METHODS: Ultrasonography fellowship-trained emergency physicians performed thoracic ultrasonography at baseline and after surgically creating a pneumothorax in eight intubated, spontaneously breathing porcine subjects. Prior to pneumothorax induction, we captured sagittal M-mode still images and B-mode videos of each intercostal space with a linear array transducer at 4cm of depth. After collection of baseline images, we placed a chest tube, injected air into the pleural space in 250mL increments, and repeated the ultrasonography for pneumothorax volumes of 250mL, 500mL, 750mL, and 1000mL. We confirmed pneumothorax with intrapleural digital manometry and ultrasound by expert sonographers. We exported collected images for interpretation by the software. We treated each individual scan as a single test for interpretation by the software. RESULTS: Excluding indeterminate results, we collected 338M-mode images for which the software demonstrated a sensitivity of 98% (95% confidence interval [CI] 92-99%), specificity of 95% (95% CI 86-99), positive likelihood ratio (LR+) of 21.6 (95% CI 7.1-65), and negative likelihood ratio (LR-) of 0.02 (95% CI 0.008-0.046). Among 364 B-mode videos, the software demonstrated a sensitivity of 86% (95% CI 81-90%), specificity of 85% (81-91%), LR+ of 5.7 (95% CI 3.2-10.2), and LR- of 0.17 (95% CI 0.12-0.22). CONCLUSIONS: This novel technology has potential as a useful adjunct to diagnose pneumothorax on thoracic ultrasonography.

Performance of Junctional Tourniquets in Normal Human Volunteers.

BACKGROUND: Inguinal bleeding is a common and preventable cause of death on the battlefield. Four FDA-cleared junctional tourniquets (Combat Ready Clamp [CRoC], Abdominal Aortic and Junctional Tourniquet [AAJT], Junctional Emergency Treatment Tool [JETT], and SAM Junctional Tourniquet [SJT]) were assessed in a laboratory on volunteers in order to describe differential performance of models. OBJECTIVE: To examine safety and effectiveness of junctional tourniquets in order to inform the discussions of device selection for possible fielding to military units. METHODS: The experiment measured safety and effectiveness parameters over timed, repeated applications. Lower extremity pulses were measured in 10 volunteers before and after junctional tourniquet application aimed at stopping the distal pulse assessed by Doppler auscultation. Safety was determined as the absence of adverse events during the time of application. RESULTS: The CRoC, SJT, and JETT were most effective; their effectiveness did not differ (p > 0.05). All tourniquets were applied safely and successfully in at least one instance each, but pain varied by model. Subjects assessed the CRoC as most tolerable. The CRoC and SJT were the fastest to apply. Users ranked CRoC and SJT equally as performing best. CONCLUSION: The CRoC and SJT were the best-performing junctional tourniquets using this model.

Initial resuscitation with plasma and other blood components reduced bleeding compared to hetastarch in anesthetized swine with uncontrolled splenic hemorrhage.

BACKGROUND: Damage control resuscitation recommends use of more plasma and less crystalloid as initial resuscitation in treating hemorrhage. The purpose of this study was to evaluate resuscitation with either blood components or conventional fluids on coagulation and blood loss. STUDY DESIGN AND METHODS: Isofluorane-anesthetized, instrumented pigs (eight per group) underwent controlled hemorrhage of 24 mL/kg, 20-minute shock period, splenic injury with 15-minute initial bleeding, and hypotensive fluid resuscitation. Lactated Ringer's (LR) was infused at 45 mL/kg while hetastarch (high-molecular-weight hydroxyethyl starch 6%, Hextend, Hospira, Inc., Lake Forest, IL) and blood component (fresh-frozen plasma [FFP], 1:1 FFP:[red blood cells] RBCs, 1:4 FFP : RBCs, and fresh whole blood [FWB]) were infused at 15 mL/kg. Postresuscitation blood loss (PRBL), hemodynamics, coagulation, hematocrit, and oxygen metabolism were measured postinjury for 5 hours. RESULTS: Resuscitation with any blood component reduced PRBL of 52% to 70% compared to Hextend, with FFP resulting in the lowest PRBL. PRBL with LR (11.5 ± 3.0 mL/kg) was not significantly different from Hextend (17.9 ± 2.5 mL/kg) or blood components (range, 5.5 ± 1.5 to 8.6 ± 2.6 mL/kg). The volume expansion effect of LR was transient. All fluids produced similar changes in hemodynamics, oxygen delivery, and demand despite the oxygen-carrying capacity of RBC-containing fluids. Compared with other fluids, Hextend produced greater hemodilution and reduced coagulation measures, which could be caused by an indirect dilutional effect or a direct hypocoagulable effect. CONCLUSIONS: These data suggest that blood products as initial resuscitation fluids reduced PRBL from a noncompressible injury compared to Hextend, preserved coagulation, and provided sustained volume expansion. There were no differences on PRBL among RBCs-to-FFP, FWB, or FFP in this nonmassive transfusion model.

Toxicity of aluminum silicates used in hemostatic dressings toward human umbilical veins endothelial cells, HeLa cells, and RAW267.4 mouse macrophages.

BACKGROUND: Aluminum silicates have been used to control bleeding after severe traumatic injury. QuikClot (QC) was the first such product, and WoundStat (WS) is the most recent. We recently observed that WS caused vascular thrombosis when applied to stop bleeding. This study investigated the cellular toxicity of WS in different cell types that may be exposed to this mineral and compared the results with other minerals such as bentonite, kaolin, and QuikClot ACS+ (QC+). METHODS: Human umbilical vein endothelial cells (HUVEC), HeLa cells, and RAW267.4 mouse macrophage-like cells (RAW) were incubated directly with different concentrations of each mineral for 24 hours. Cell viability was determined metabolically using the AlamarBlue fluorescent technique. In another experiment, minerals were exposed to HUVEC via Transwell inserts with a polycarbonate filter (0.4-µm pore size) to prevent direct contact between cells and minerals for determining whether direct exposure or leaching compounds from minerals cause cytotoxicity. RESULTS: Incubation of HUVEC and RAW cells with 1 to 100 µg/mL of the minerals for 24 hours resulted in differential toxicities. The cytotoxicity of WS was equal to that of bentonite and higher than kaolin and QC+. Neither cell type survived for 24 hours in the presence of 100 µg/mL WS or bentonite. These minerals, however, had little effect on the viability of HeLa cells. In the second HUVEC experiment, a 10 times higher concentration of these compounds placed in Transwell inserts yielded no decrease in cell viability. This result indicates that leaching toxicants or binding of nutrients by the ion-exchange properties of minerals did not cause the toxicity. CONCLUSIONS: Although aluminum silicates seem relatively innocuous to epithelial cells, all produced some toxicity toward endothelial cells and macrophages. WS and bentonite were significantly more toxic than kaolin and zeolite present in QC+, respectively, at equivalent doses. The cytotoxic effect seemed to be caused by the direct contact of the minerals with the cells present in wounds. These data suggest that the future clearance of mineral-based hemostatic agents should require more extensive cytotoxicity testing than the current Food and Drug Administration requirements.

Coagulation changes to systemic acidosis and bicarbonate correction in swine.

BACKGROUND: As part of our overall interest in the mechanisms and treatment related to the development of the lethal triad of hypothermia, acidosis, and coagulopathy seen in trauma patients, the purpose of this study was to determine whether acidosis, inducible either by HCl infusion or hemorrhage/hypoventilation, leads to coagulopathy, and if correction of the acidosis will alleviate this coagulopathy. METHODS: In two separate experiments, acidosis was induced in anesthetized swine by (1) HCl infusion (n = 10) or (2) hemorrhage/hypoventilation (n = 8). Arterial blood samples were taken before HCl infusion or hemorrhage (arterial pH 7.4), after HCl infusion or hemorrhage (pH 7.1), and after bicarbonate infusion to return pH to 7.4. Arterial pH and blood gases were measured every 15 minutes. RESULTS: Acidosis (arterial pH 7.1) led to a hypocoagulation as measured by several coagulation parameters. In both experiments, acidosis was associated with a significant decrease in the maximum strength of the clot and the rate at which the clot formed. There was a significant decrease in endogenous thrombin potential and maximum thrombin concentration after acidosis in both groups (thrombin generation assay). However, the activated clotting time, prothrombin time, and activated partial thromboplastin time were significantly elevated only in the HCl-infused group. Fibrinogen concentration and platelet count were significantly reduced in both groups after acidosis. The hypocoagulation that was induced by either hemorrhage/hypoventilation or HCl infusion was not immediately corrected after returning pH to 7.4 with bicarbonate injection. CONCLUSIONS: These data suggest that acidosis induced by HCl infusion or by hemorrhage/hypoventilation leads to hypocoagulation. Simple correction of the arterial pH with bicarbonate is not sufficient to correct this coagulopathy.

Development of a standard swine hemorrhage model for efficacy assessment of topical hemostatic agents.

BACKGROUND: The diverse information of efficacy of hemostatic products, obtained from different military laboratories using different models, has made it difficult to ascertain the true benefit of new hemostatic agents in military medicine. The aim of this study was to recommend a standard hemorrhage model for efficacy testing acceptable by most investigators in the field and avoid contradictory and duplicative efforts by different laboratories. METHODS: The swine femoral artery injury model (6-mm arteriotomy) with some modifications was tested to standardize the model. The suggested modifications included no splenectomy, one-time treatment, 30 seconds free bleeding, and 5 L limit for fluid resuscitation. The model was tested with all or some of these modifications in four experimental conditions (n = 5-6 pigs per condition) using Combat Gauze (CG) as control agent. RESULTS: The primary end points including blood pressure, blood loss, and survival rates were modestly changed in the four conditions. The second experimental condition in which bleeding was treated with a single CG with 3-minute compression produced the most suitable results. The average blood loss was 99 mL/kg, and hemostasis was achieved in one-third of the pigs, which led to matching survival rate. CONCLUSION: A rigorous hemorrhage model was developed for future evaluation of new hemostatic agents and comparison with CG, the current standard of care. This model may not be suitable for testing every agent and some modifications may be necessary for specific applications. Furthermore, laboratory studies using this or similar models must be accompanied by operational testing in the field to confirm the efficacy and practical utility of selected agents when used on the battlefield.

Effect of hemodilution on coagulation and recombinant factor VIIa efficacy in human blood in vitro.

BACKGROUND: This study evaluates the effect of hemodilution by various common resuscitation fluids, and the efficacy of activated recombinant factor VII (rFVIIa) on coagulation parameters in human blood in vitro. METHODS: Samples from normal healthy volunteers (n = 9) were hemodiluted from 0% to 90% with normal saline, or 0%, 40%, 60%, and 80% with 5% albumin, Hespan, Hextend, normal saline, or lactated Ringer's, and incubated at 37°C ± 1°C for 30 minutes with and without rFVIIa (1.26 µg/mL). RESULTS: There was a strong correlation between the dilution of hemoglobin (Hb), platelets, or fibrinogen and coagulation parameters. Hemodilution 0% to 90% changed coagulation parameters (prothrombin time [PT], activated partial thromboplastin time [aPTT], and thromboelastography) in an exponential fashion; the greatest changes occurred after hemodilution lowered Hb <6 mg/dL, platelet count < 100,000/mm(3), and fibrinogen concentration <200 mg/dL. PT and aPTT were significantly prolonged after 60% and 80% dilution for all fluids. Hemodilution of 60% and 80% significantly decreased clot strength (maximum amplitude) and the kinetics of clot development (α angle) and increased the clot formation time (K). Hemodilution with Hextend and Hespan decreased maximum amplitude and α angle >5% albumin, lactated Ringer's, or normal saline. rFVIIa significantly improved PT at 60% and 80% dilutions, and aPTT at 80% dilution. There was a significant effect of dilution, but not fluid type, on the efficacy of rFVIIa to change PT and aPTT, and the onset of clotting (R). CONCLUSIONS: We have strong in vitro evidence that Hb <6 mg/dL, platelet count <100,000/mm(3), and fibrinogen concentration <200 mg/dL can be used as indexes of hemodilution-induced coagulopathy. This study also shows that Hextend and Hespan tend to decrease the clotting ability >5% albumin or the crystalloids. rFVIIa significantly decreased PT at all dilutions and aPTT at the highest dilution. The effectiveness of rFVIIa on PT and aPTT was significantly affected by the degree of dilution, but not by the type of fluid.

Evaluation of a novel hydrogel intravascular embolization agent in a swine model of fatal uncontrolled solid organ hemorrhage and coagulopathy.

INTRODUCTION: Current agents for the intravascular embolization of traumatic hemorrhage are used off-label and have been minimally studied with respect to their performance under differing coagulation conditions. We studied the hemorrhage control efficacy of a novel, liquid, polyethylene glycol-based hydrogel delivered as two liquid precursors that polymerize within the target vessel in a unique animal model of severe solid organ injury with and without dilutional coagulopathy. METHODS: Anesthetized swine (n = 36, 45 ± 3 kg) had laparotomy and splenic externalization. Half underwent 50% isovolemic hemodilution with 6% hetastarch and cooling to 33°C-35°C (coagulopathic group). All animals had controlled 20 mL/kg hemorrhage and endovascular proximal splenic artery access with a 4F catheter via a right femoral sheath. Splenic transection and 5-minute free bleeding were followed by treatment (n = 5/group) with 5 mL of gelfoam slurry, three 6-mm coils, up to 6 mL of hydrogel, or no treatment (n = 3, control). Animals received 15 mL/kg plasma and were monitored for 6 hours with continuous blood loss measurement. RESULTS: Coagulopathy was successfully established, with coagulopathic animals having greater pretreatment blood loss and earlier mean time to death regardless of the treatment group. All control animals died within 100 minutes. Overall survival without coagulopathy was 5/5 for hydrogel, 4/5 for coil, and 3/5 for gelfoam. With coagulopathy, one hydrogel animal survived to the end of the experiment, with 2/4 hydrogel deaths occurring in the final hour of observation. In noncoagulopathic animals, hydrogel demonstrated improved survival time (P < .01) and post-treatment blood loss (1.46 ± 0.8 mL/kg) over controls (18.8 ± 0.7, P = .001), gelfoam (4.7 ± 1.3, P > .05), and coils (4.6 ± 1.5, P > .05). In coagulopathic animals, hydrogel had improved survival time (P = .003) and decreased blood loss (4.2 ± 0.8 mL/kg) compared with control (20.4 ± 4.2, P = .003). CONCLUSIONS: The hydrogel demonstrated equivalent hemorrhage control performance to standard treatments under noncoagulopathic conditions and improved performance in the face of dilutional coagulopathy. This agent should be explored as a potential preferable treatment for the embolization of traumatic solid organ and other injuries. (JVS-Vascular Science 2021;2:43-51.). CLINICAL RELEVANCE: In a translational model of severe solid organ injury hemorrhage with and without coagulopathy, a novel hydrogel transarterial embolization agent demonstrated equivalent hemorrhage control performance to standard agents under noncoagulopathic conditions and improved performance in the face of dilutional coagulopathy. This agent represents a promising future treatment for the embolization of traumatic solid organ and other injuries.

Safety evaluation of new hemostatic agents, smectite granules, and kaolin-coated gauze in a vascular injury wound model in swine.

BACKGROUND: In 2007, a potent procoagulant mineral called WoundStat (WS), consisting of smectite granules, received clearance from the Food and Drug Administration for marketing in the United States for temporary treatment of external hemorrhage. Previously, we found that microscopic WS particles remained in the injured vessels that were treated, despite seemingly adequate wound debridement. Thus, we investigated the thromboembolic risk of using WS when compared with kaolin-coated gauze, Combat Gauze (CG); or regular gauze, Kerlix (KX) to treat an external wound with vascular injuries in pigs. METHODS: The right common carotid artery and external jugular vein of pigs were isolated and sharply transected (50%). After 30 seconds of free bleeding, the neck wounds were packed with WS, CG, or KX and compressed until hemostasis was achieved (n = 8 per group). Wounds were debrided after 2 hours, and vascular injuries were primarily repaired with suture. Blood flow was restored after infusing 1 L of crystalloid (no heparin or aspirin) and the wounds were closed. Two hours later, computed tomographic angiography was performed, and the wounds were reopened to harvest the vessels. The brains and lungs were recovered for gross and microscopic examination after euthanasia. RESULTS: No differences were found in baseline measurements. Thrombelastography showed similar hypercoagulability of the final blood samples when compared with baselines in all groups. All vessels treated with KX or CG were patent and had no thrombus or blood clot in their lumen. In contrast, seven of eight carotid arteries and six of eight jugular veins treated with WS developed large occlusive red thrombi and had no flow. Small clots and WS residues were also found in the lungs of two pigs. Histologically, significant endothelial and transmural damage was seen in WS-treated vessels with luminal thrombi and embedded WS residues. CONCLUSION: WS granules caused endothelial injury and significant transmural damage to the vessels that render them nonviable for primary surgical repair. The granules can enter systemic circulation and cause distal thrombosis in vital organs. More relevant in vitro and in vivo safety tests should be required for clearance of new hemostatic agents.

Abdominal aortic and junctional tourniquet versus zone III resuscitative endovascular balloon occlusion of the aorta in a swine junctional hemorrhage model.

BACKGROUND: Junctional hemorrhage is a leading contributor to battlefield mortality. The Abdominal Aortic and Junctional Tourniquet (AAJT) and infrarenal (zone III) resuscitative endovascular balloon occlusion of the aorta (REBOA) are emerging strategies for controlling junctional hemorrhage, with AAJT currently available in select forward deployed settings and increasing interest in applying REBOA in the military prehospital environment. This study compared the hemostatic, hemodynamic, and metabolic effects of these devices used for junctional hemorrhage control. METHODS: Shock was induced in anesthetized, mechanically ventilated swine with a controlled hemorrhage (20 mL/kg) and closed femur fracture followed by uncontrolled hemorrhage from a partial femoral artery transection (40% total hemorrhage volume). Residual femoral hemorrhage was recorded during 60-minute AAJT (n = 10) or zone III REBOA (n = 10) deployment, and the arterial injury was repaired subsequently. Animals were resuscitated with 15 mL/kg autologous whole blood and observed for 6 hours. RESULTS: One animal in each group died during observation. Both devices achieved hemostasis with mean residual femoral blood loss in the AAJT and REBOA groups of 0.38 ± 0.59 mL/kg and 0.10 ± 0.07 mL/kg (p = 0.16), respectively, during the 60-minute intervention. The AAJT and REBOA augmented proximal blood pressure equally with AAJT allowing higher distal pressure than REBOA during intervention (p < 0.01). Following device deflation, AAJT animals had transiently lower mean arterial blood pressure than REBOA pigs (39 ± 6 vs. 54 ± 11 mm Hg p = 0.01). Both interventions resulted in similar degrees of lactic acidemia which resolved during observation. Similar cardiac and renal effects were observed between AAJT and REBOA. CONCLUSION: The AAJT and REBOA produced similar hemostatic, resuscitative, and metabolic effects in this model of severe shock with junctional hemorrhage. Both interventions may have utility in future military medical operations.

Should Albumin be Considered for Prehospital Resuscitation in Austere Environments? A Prospective Randomized Survival Study in Rabbits.

BACKGROUND: The new guidelines for prehospital care of combat casualties in shock recommend administration of whole blood or blood components to increase blood pressure to a permissible hypotensive level (i.e., hypotensive resuscitation [HR]). We investigated if 2 h of HR using limited volumes of whole blood, plasma, or albumin would lead to full recovery and long-term survival of rabbits subjected to severe hemorrhagic shock (HS). METHODS: Following instrumentation, laparotomy was performed on IV-anesthetized spontaneously breathing New Zealand white rabbits (3.0 kg -3.5 kg). Next, ∼40% of rabbits' blood volume was removed producing HS (mean arterial pressure [MAP]∼20 mm Hg). Fifteen minutes later, rabbits were resuscitated with a limited volume (12.5 mL/kg) of rabbit whole blood (fresh whole blood [FWB]), rabbit fresh frozen plasma (FFP), or 5% human albumin (ALB) to a target pressure (MAP) of 60 mm Hg (n=8/grp) and monitored for 2 h. Liver bleeding time was measured at baseline and 10 min after HR. Subsequently, animals were fully resuscitated (blood + lactated Ringer [LR]), surgically repaired, and recovered for 8 days. An untreated group (n = 6) was also included. RESULTS: Following HS, lactate and base deficit levels were increased to 8.2 ±â€Š1.6 and 12.9 ±â€Š3.1 mM respectively with no difference among groups. A lower volume of FWB volume was required to reach the target MAP (P < 0.05 vs. ALB) but MAP declined during the HR period (P < 0.01 vs. ALB). FWB provided higher hematocrit and platelets but it did not reduce lactate level faster than other fluids. Beside higher fibrinogen, no differences were found in hemostatic or resuscitative effects of FFP versus ALB. Bleeding time was prolonged with ALB and FFP fluids but unchanged with FWB. Untreated rabbits died during shock or shortly after. All treated rabbits except one recovered and lived for 8 days with normal blood tests and similar tissue histology. CONCLUSIONS: Two hours of HR using a limited volume of FWB, FFP, or ALB led to full recovery and long-term survival of rabbits subjected to HS. Apart from bleeding time, no clinically significant differences were found among the three fluids. Five percent human albumin solutions are isotonic, iso-oncotic, ready-to-use, stable, and compatible with all blood types and should be considered for prehospital resuscitation where blood products are not available or not accepted.

Determination of efficacy of new hemostatic dressings in a model of extremity arterial hemorrhage in swine.

BACKGROUND: The HemCon (HC) bandage and QuickClot have been used over the past 6 years for treating external compressible hemorrhage in combat casualties. Previously, we tested three new hemostatic agents in granular/powder forms that were superior to these products. In this study, four new dressings (preselected) that are more suitable for battlefield application were evaluated. The efficacy and acute safety of the dressings were tested in our standard arterial hemorrhage model. METHODS: Anesthetized pigs (n = 38, 37 kg) were instrumented, and arterial blood was collected for hematological and coagulation assays. After splenectomy, the right femoral artery was isolated, injured (6 mm arteriotomy), and unrestricted bleeding allowed for 45 seconds. A hemostatic dressing (HC RTS [n = 6], Celox-D [CXb, n = 6], TraumaStat [TS, n = 10], Combat Gauze [CG, n = 10], or placebo gauze [PG, n = 6]) was then applied over the wound randomly and compressed for 2 minutes. Fluid resuscitation was administered and titrated to maintain a mean arterial pressure of 65 mm Hg. Animals were observed for 180 minutes or until death. Computed tomography angiography was performed on survivors and tissues were collected for histology. RESULTS: No differences were found in baseline blood measures, pretreatment blood loss or fluid infusion among groups. HCs and CXb testing discontinued after six unsuccessful tests, and the data were excluded. Stable hemostasis was achieved in two PG, two TS, and eight CG pigs in remaining groups resulting in stabilized mean arterial pressure and significantly different survival rates (20-80%, p = 0.03). CG secured hemostasis for 134.6 minutes +/- 22.2 minutes, which was significantly longer than TS (35.7 +/- 22.0 minutes, p < 0.05) but not different from PG (57.9 +/- 36.2 minutes). The average survival time of CG-treated animals (167.3 +/- 5.9 minutes) was also significantly longer (p < 0.05) than that of TS- (90.0 +/- 15.3 minutes) or PG-treated (121 +/- 19.3 minutes) pigs. Posttreatment blood loss was less in CG (37.4 +/- 17.3 mL/kg) than that of the two other groups (TS = 79.8 +/- 13.8 mL/kg and PG = 75.5 +/- 23.8 mL/kg), but this difference was not significant. No significant rise in wound temperature (>1 degrees C) was recorded after treatment with dressings and computed tomography images showed no flow through the vessels. Histologic observations showed mild to moderate changes in treated vessels with no difference between CG and PG. In vitro analysis of blood treated with CG or PG (lesser extent) showed increased clotting rate and clot strength. TS treatment had no effect on blood clotting activity. CONCLUSION: CG was the most effective dressing tested in this arterial hemorrhage model. The hemostatic property of CG is attributed to its raw material (nonwoven Rayon and polyester blend), kaolin coating, and the large surface area (3 inch x 4 yd) of this absorbent sponge. CG is now recommended as the first line of treatment for life-threatening hemorrhage on the battlefield, replacing HC.

Comparison of new hemostatic granules/powders with currently deployed hemostatic products in a lethal model of extremity arterial hemorrhage in swine.

BACKGROUND: HemCon bandage (HC) and QuikClot granules (QC) have been deployed for the past 5 years for treating external hemorrhage in combat casualties. We examined efficacy and initial safety of three new hemostatic granules/powders in a swine extremity arterial hemorrhage model that was 100% fatal with army standard gauze treatment. The new products were compared with the most advanced forms of HC and QC products. METHODS: Anesthetized pigs (37 kg, n = 46) were instrumented, splenectomized, and their femoral arteries were isolated and injured (6 mm arteriotomy). After 45 seconds free bleeding, a test agent [WoundStat (WS), super quick relief (SQR), Celox (CX)] or a control product [HC or QC bead bags (advanced clotting sponge plus)] was applied to the wounds and compressed with a large gauze for 2 minutes. Fluid resuscitation (colloid and crystalloid) was given and titrated to a mean arterial pressure of 65 mm Hg. Animals were observed for 180 minutes or until death. Computed tomography angiography was performed on survivors and tissue samples were collected form wounds for histologic examination. RESULTS: No differences were found in baseline measurements and pretreatment blood loss (17.4 mL/kg +/- 0.5 mL/kg, mean +/- SEM) among groups. Advanced clotting sponge plus testing was halted after six unsuccessful attempts (no hemostasis observed) whereas other agents were tested each in 10 animals. Stable hemostasis was achieved in 10 (WS), 7 (SQR), 6 (CX), and 1 (HC) subjects in each group, resulting in the recovery of mean arterial pressure and survival of the animals for 3 hours (p < 0.05, SQR or WS vs. HC). Posttreatment blood loss was significantly reduced with the use of the new agents (CX = 40 +/- 16.6, SQR = 34.5 +/- 16.3, WS = 9.5 mL/kg +/- 5.2 mL/kg) as compared with HC (85.6 mL/kg +/- 10 mL/kg, p < 0.05). The granular treated animals lived for 180 (WS), 164 +/- 8.2 (SQR) and 138 +/- 17.7 (CX) minutes, significantly (p < 0.05) longer than the HC (83.3 +/- 12 minutes) group. A significant (p < 0.05) rise in temperature (53.5 degrees C +/- 1.8 degrees C) over baseline (36.5 degrees C +/- 0.3 degrees C) was measured only in the wounds treated with SQR. Computed tomography images showed no blood flow through treated vessels. Histologic evidence indicated the least tissue damage with HC, moderate damage with WS and CX, and most damage including axonal necrosis with SQR. CONCLUSION: The new hemostatic agents are significantly more effective in treating arterial hemorrhage than currently deployed products. Among them, WS granules appear to be most efficacious, followed by SQR and CX powders. The clinical significance of tissue damage caused by these agents and any potential risk of embolism with procoagulant granular/powder products are unknown and warrant survival studies.

Thromboelastography as a better indicator of hypercoagulable state after injury than prothrombin time or activated partial thromboplastin time.

OBJECTIVES: To investigate the hemostatic status of critically ill, nonbleeding trauma patients. We hypothesized that a hypercoagulable state exists in patients early after severe injury and that the pattern of clotting and fibrinolysis are similar between burned and nonburn trauma patients. MATERIALS: Patients admitted to the surgical or burn intensive care unit within 24 hours after injury were enrolled. Blood samples were drawn on days 0 through 7. Laboratory tests included prothrombin time (PT), activated partial thromboplastin time (aPTT), levels of activated factor XI, D-dimer, protein C percent activity, antithrombin III percent activity, and thromboelastography (TEG). RESULTS: Study subjects were enrolled from April 1, 2004, to May 31, 2005, and included nonburn trauma patients (n = 33), burned patients (n = 25), and healthy (control) subjects (n = 20). Despite aggressive thromboprophylaxis, three subjects (2 burned and 1 nonburn trauma patients [6%]) had pulmonary embolism during hospitalization. Compared with controls, all patients had prolonged PT and aPTT (p < 0.05). The rate of clot formation (alpha angle) and maximal clot strength were higher for patients compared with those of controls (p < 0.05), indicating a hypercoagulable state. Injured patients also had lower protein C and antithrombin III percent activities and higher fibrinogen levels (p < 0.05 for all). Activated factor XI was elevated in 38% of patients (control subjects had undetectable levels). DISCUSSION: Thromboelastography analysis of whole blood showed that patients were in a hypercoagulable state; this was not detected by plasma PT or aPTT. The high incidence of pulmonary embolism indicated that our current prophylaxis regimen could be improved.

Effect of partial and complete aortic balloon occlusion on survival and shock in a swine model of uncontrolled splenic hemorrhage with delayed resuscitation.

BACKGROUND: Resuscitative endovascular balloon occlusion of the aorta (REBOA) is accepted as a resuscitation adjunct and bridge to definitive hemostasis. The ischemic burden of REBOA may be mitigated by a partial REBOA (P-REBOA) strategy permitting longer occlusion times and military use for combat trauma. We evaluated REBOA and P-REBOA in a swine multiple trauma model with uncontrolled solid organ hemorrhage and delayed resuscitation and surgical hemostasis. METHODS: Anesthetized swine (51.9 ± 2.2 kg) had 20 mL/kg hemorrhage and closed femur fracture. Splenic transection was performed and free bleeding permitted for 10 minutes. Controls (n = 5) were hemorrhaged but had no REBOA, REBOA (n = 8) had 60 minutes complete zone 1 occlusion, P-REBOA (n = 8) had 15 minutes complete occlusion and 45 minutes 50% occlusion. Splenectomy was performed and plasma (15 mL/kg) resuscitation initiated 5 minutes prior to deflation. Resuscitation goal was 80 mm Hg systolic with epinephrine as needed. Animals were monitored for 6 hours. RESULTS: An initial study with 120-minute occlusion had universal fatality in three REBOA (upon deflation) and three P-REBOA animals (after 60 minutes inflation). With 60-minute occlusion, mortality was 100%, 62.5%, and 12.5% in the control, REBOA, and P-REBOA groups, respectively (p < 0.05). Survival time was shorter in controls (120 ± 89 minutes) than REBOA and P-REBOA groups (241 ± 139, 336 ± 69 minutes). Complete REBOA hemorrhaged less during inflation (1.1 ± 0.5 mL/kg) than Control (5.6 ± 1.5) and P-REBOA (4.3 ± 1.4), which were similar. Lactate was higher in the REBOA group compared with the P-REBOA group after balloon deflation, remaining elevated. Potassium increased in REBOA after deflation but returned to similar levels as P-REBOA by 120 minutes. CONCLUSION: In a military relevant model of severe uncontrolled solid organ hemorrhage 1-hour P-REBOA improved survival and mitigated hemodynamic and metabolic shock. Two hours of partial aortic occlusion was not survivable using this protocol due to ongoing hemorrhage during inflation. There is potential role for P-REBOA as part of an integrated minimally invasive field-expedient hemorrhage control and resuscitation strategy.

A novel biologic hemostatic dressing (fibrin patch) reduces blood loss and resuscitation volume and improves survival in hypothermic, coagulopathic Swine with grade V liver injury.

OBJECTIVES: This study evaluated the efficacy of a biologic hemostatic fibrin patch (FP) to control coagulopathic bleeding and prevent death in a porcine model of severe liver injury with hemodilution and hypothermia. METHODS: Coagulopathy was produced in swine by exchanging 60% of the animals' circulating blood volume with Hextend and lowering the core temperature to 32.0 degrees C +/- 0.5 degrees C. A grade V liver injury was induced and allowed to bleed freely for 30 seconds (pretreatment blood loss). Animals were randomly divided into three treatment groups: hepatic packing (HP) using laparotomy sponges, FP application plus HP, or placebo patch (PP) application plus HP. Animals were resuscitated to 80% of the preinjury mean arterial pressure. Core temperature, mean arterial pressure, and survival were monitored for 1 hour postinjury. Packs were removed from the animals that survived to 1 hour and they were monitored for an additional hour. RESULTS: Coagulopathy was confirmed by significant increases (p < 0.01) in prothrombin time, activated partial thromboplastin time, and activated clotting time in preinjury measurements as compared with baseline values. Pretreatment blood loss was not different among the groups. However, significant (p < 0.01) differences were observed in the posttreatment blood loss (772 mL +/- 340 mL, 4,977 mL +/- 440 mL, 4,173 mL +/- 608 mL), as well as the required fluid resuscitation volume (994 mL +/- 26 mL, 4,083 mL +/- 185 mL, 3,494 mL +/- 492 mL), between FP versus PP or HP groups, respectively. In addition, 89% of FP animals survived the 2-hour observation with an average survival time of 111 minutes +/- 9 minutes, which was significantly higher than the PP (0% survival, 39 minutes +/- 4 minutes) or HP (13% survival, 41 minutes +/- 12 minutes) groups. CONCLUSION: FP with packing effectively controlled coagulopathic bleeding and prevented death in a model of grade V liver injury in which HP alone (standard of care) was ineffective.