Porcine Model Of Septic Shock Research Articles

Hyaluronic acid plasma levels during high versus low tidal volume ventilation in a porcine sepsis model.

BackgroundShedding of the endothelial glycocalyx can be observed regularly during sepsis. Moreover, sepsis may be associated with acute respiratory distress syndrome (ARDS), which requires lung protective ventilation with the two cornerstones of application of low tidal volume and positive end-expiratory pressure. This study investigated the effect of a lung protective ventilation on the integrity of the endothelial glycocalyx in comparison to a high tidal volume ventilation mode in a porcine model of sepsis-induced ARDS.MethodsAfter approval by the State and Institutional Animal Care Committee, 20 male pigs were anesthetized and received a continuous infusion of lipopolysaccharide to induce septic shock. The animals were randomly assigned to either low tidal volume ventilation, high tidal volume ventilation, or no-LPS-group groups and observed for 6 h. In addition to the gas exchange parameters and hematologic analyses, the serum hyaluronic acid concentrations were determined from central venous blood and from pre- and postpulmonary and pre- and postcerebral circulation. Post-mortem analysis included histopathological evaluation and determination of the pulmonary and cerebral wet-to-dry ratios.ResultsBoth sepsis groups developed ARDS within 6 h of the experiment and showed significantly increased serum levels of hyaluronic acid in comparison to the no-LPS-group. No significant differences in the hyaluronic acid concentrations were detected before and after pulmonary and cerebral circulation. There was also no significant difference in the serum hyaluronic acid concentrations between the two sepsis groups. Post-mortem analysis showed no significant difference between the two sepsis groups.ConclusionIn a porcine model of septic shock and ARDS, the serum hyaluronic acid levels were significantly elevated in both sepsis groups in comparison to the no-LPS-group. Intergroup comparison between lung protective ventilated and high tidal ventilated animals revealed no significant differences in the serum hyaluronic acid levels.

Effects of ulinastatin on renal perfusion evaluated by Doppler ultrasonography in a porcine model of septic shock.

The present study aimed to evaluate the effect of ulinastatin (UTI) on renal perfusion using Doppler ultrasonography in a porcine model of septic shock induced by smoking inhalation and live methicillin-resistant Staphylococcus aureus instillation. A total of 32 healthy Landrace pigs were randomly assigned into the following four groups: Sham group (SH; n=5), septic shock group (SS; n=9), septic shock treated with vancomycin (15 mg/kg) group (VAN; n=9) and septic shock treated with UTI (50,000 U/kg) + vancomycin (UTI; n=9) group. Renal perfusion was evaluated by contrast-enhanced ultrasound (CEUS) at baseline and at the end of the protocol (24 h). The spectrum of interlobar or arcuate artery was selected to calculate the corrected resistive index (cRI). Sulphur hexafluoride microbubbles were bolus injected via a venous catheter. The peak intensity (Pi) and area under curve (AUC) were calculated using a time-intensity curve. Compared with the baseline group, cRI was increased significantly at the end of the protocol, except for that in the SH group, whereas Pi decreased significantly after injury in all experimental groups but was higher in the UTI group compared with that in the SS and VAN groups (both P<0.001). Linear correlation was found between the cardiac output (CO) and Pi (R2=0.752; P<0.001). The AUC was significantly decreased after injury in the SS and VAN groups compared with the baseline group. All parameters detected by CEUS were improved in the UTI group, and significant differences were found between the UTI and SS or VAN group (all P<0.05). In conclusion, acute renal injury, which occasionally occurs during septic shock, is accompanied with a significantly lower perfusion rate in the renal microcirculation. By contrast, UTI can significantly improve renal perfusion, which can be reliably evaluated using CEUS.

MicroTools enables automated quantification of capillary density and red blood cell velocity in handheld vital microscopy

Direct assessment of capillary perfusion has been prioritized in hemodynamic management of critically ill patients in addition to optimizing blood flow on the global scale. Sublingual handheld vital microscopy has enabled online acquisition of moving image sequences of the microcirculation, including the flow of individual red blood cells in the capillary network. However, due to inherent content complexity, manual image sequence analysis remained gold standard, introducing inter-observer variability and precluding real-time image analysis for clinical therapy guidance. Here we introduce an advanced computer vision algorithm for instantaneous analysis and quantification of morphometric and kinetic information related to capillary blood flow in the sublingual microcirculation. We evaluated this technique in a porcine model of septic shock and resuscitation and cardiac surgery patients. This development is of high clinical relevance because it enables implementation of point-of-care goal-directed resuscitation procedures based on correction of microcirculatory perfusion in critically ill and perioperative patients.

A Novel Porcine Model of Septic Shock Induced by Acute Respiratory Distress Syndrome due to Methicillin-resistant Staphylococcus aureus

Background:Sepsis is one of the main causes of mortality in critically ill patients following progression to septic shock. To investigate the pathophysiologic changes of sepsis, we developed a novel porcine model of septic shock induced by acute respiratory distress syndrome (ARDS) due to methicillin-resistant Staphylococcus aureus (MRSA) pneumonia.Methods:Twenty-six male Landraces (Lvyuanweiye, Beijing, China) weighing 30 ± 2 kg were divided into four groups: sham group (SH; n = 5); cotton smoke inhalation group (SM; n = 6); MRSA pneumonia group (MR; n = 6); and septic shock group with cotton smoke inhalation + MRSA pneumonia (SS; n = 9). Extensive hemodynamics, oxygen dynamics, and lung function were monitored for 24 h following the injury or until death. Tissues were collected, and histopathology evaluations were carried out.Results:Blood cultures from 6 of 9 animals in the SS group were positive for MRSA. Two hours following the injury, decreased mean arterial blood pressure (60–70 mmHg) and cardiac index (<2 L·min−1·m−2) were observed in the animals in the SS group, while systemic vascular resistance index was increased. The hemodynamic characteristics of septic shock were only observed in the SS group but not significant in the other groups. The PO2/FiO2 in the SM and SS groups decreased to 300 and 100, respectively. In the SS group, extravascular lung water index increased to 20 ml/kg, whereas thoracopulmonary compliance decreased to 10 ml/H2O after injury. Deterioration of pulmonary function in the SS group was more serious than the SM and MR groups. Severe lung injury in the SS group was confirmed by the histopathology evaluations. The lung injury confirmed by high-resolution thin-section computed tomography and histopathology in the SS group was more serious than those of other groups.Conclusions:In the present study, we developed a novel porcine model of septic shock induced by ARDS due to severe MRSA pneumonia with characteristic hyperdynamic and hypodynamic phases in 24 h, which mimicked the hemodynamic changing of septic shock in human.

Ulinastatin ameliorates gastrointestinal injury sustained in a 2-hit porcine model of septic shock

BackgroundUlinastatin is protective against organ dysfunction in severe disease. We investigated the extent of gastrointestinal tract injury and the potential protective effect of ulinastatin in a 2-hit porcine model of septic shock. MethodsPigs were randomized to 4 groups, 3 septic shock groups (12 per group)—vancomycin (VAN), vancomycin + ulinastatin (VAN + ULI), and saline (SAL)—and a sham-operated group (n = 10). Septic shock was induced by 2 hits: acute lung injury and Staphylococcus aureus challenge. Four hours after the 2 hits, pigs in septic shock received a central venous injection of vancomycin, vancomycin + ulinastatin, or saline. Hemodynamic status and blood samples were obtained. Serum d-lactate, diamine oxidase, and intestinal fatty acid–binding protein were determined, and gastrointestinal ATP enzyme activity was measured. Pathological and ultrastructural tests were performed. ResultsGastrointestinal tract injury after septic shock was significant. Compared with the SAL and VAN groups, the VAN + ULI group had better hemodynamic parameters (improved mean arterial pressure and cardiac output) (P< .05) and improved oxygen metabolism (oxygen delivery and consumption) (P< .05). In VAN + ULI group, serum d-lactate, diamine oxidase, and intestinal fatty acid–binding protein were significantly reduced (P< .05). Moreover, Na+-K+- and Ca2+-ATPase enzyme activity was significantly high (P< .05). Pathological and ultrastructural changes showed that severe gastrointestinal injury was significantly ameliorated in the VAN + ULI group vs the SAL and VAN groups. ConclusionsGastrointestinal injury and abnormal energy metabolism are remarkable following septic shock. Ulinastatin can improve energy metabolism and ameliorate injury to the gastrointestinal mucosa in the early stage of septic shock.

Renal haemodynamic response to amino acids infusion in an experimental porcine model of septic shock.

Acute kidney injury (AKI) is common in sepsis. Treatments allowing maintenance of renal blood flow (RBF) could help to prevent AKI associated with renal hypoperfusion. Amino acids (AA) have been associated with an increase of RBF and glomerular filtration rate (GFR) in several species. The aim of this study was to evaluate the effects of an AA infusion on RBF and GFR in a porcine model of septic shock. A total of 17 piglets were randomly assigned into three groups: Sham (Sham, n = 5), sepsis without AA (S-NAA, n = 6), sepsis treated with AA (S-AA, n = 6). Piglets preparation included the placement of ultrasonic transit time flow probes around left renal artery for continuous RBF measurement; ureteral catheters for GFR and urine output evaluation; pulmonary artery catheter for cardiac output (CO) and pulmonary arterial pressure measurements. Mean arterial pressure (MAP) and renal vascular resistance (RVR) were also determined. Septic shock was induced with a live Pseudomonas aeruginosa infusion. Crystalloids, colloids and epinephrine infusion were used to maintain and restore MAP > 60 mmHg and CO > 80% from baseline. Renal haemodynamic did not change significantly in the Sham group, whereas RBF increased slightly in the S-NAA group. Conversely, a significant increase in RVR and a decrease in RBF and GFR were observed in the S-AA group. AA infusion was associated with a higher requirement of epinephrine [340.0 (141.2; 542.5) mg vs. 32.5 (3.8; 65.0) mg in the S-NAA group P = 0.044]. An infusion of amino acids impaired renal haemodynamics in this experimental model of septic shock.

Intraosseous and intravenous administration of antibiotics yields comparable plasma concentrations during experimental septic shock.

BackgroundWe aimed to investigate whether comparable antibiotic concentrations could be reached with intraosseous and intravenous administration during septic shock.MethodsIn this randomized, prospective experimental study conducted at an animal research laboratory at the University Hospital of Uppsala, eight anesthetized pigs, weighing 21.2 to 29.1 kg (mean: 25.2 ± 2.3 kg), received endotoxin infusion at 4 μg/kg/h for 6 h. At the onset of clinical shock, alternatively after 3 h of endotoxemia, they received 75 mg/kg of cefotaxime and 7 mg/kg of gentamicin either in a proximal tibial intraosseous catheter or in a peripheral intravenous catheter. Mixed venous samples were taken after 5, 15, 30, 60, 120 and 180 min and analyzed for antibiotic concentrations.ResultsFor both antibiotics, plasma concentrations after intraosseous and intravenous administration followed similar curves throughout the observation period, and peak concentrations were comparable. Mean concentration area under the curve (AUC mg × h/l) for cefotaxime was 108.1 ± 19.5 after intraosseous and 116.5 ± 11.1 after intravenous administration; ratio 0.93, (95% CI 0.71–1.19). Mean AUC for gentamicin was 28.1 ± 6.8 for intraosseous and 32.2 ± 3.5 for intravenous administration; ratio 0.87 (95% CI 0.62–1.19).ConclusionsIn this porcine septic shock model, intraosseous and intravenous administration of gentamicin and cefotaxime yielded comparable concentrations. In an emergency, intraosseous administration of these antibiotics may be considered in severe infections when venous access is difficult.

A bio-artificial renal epithelial cell system conveys survival advantage in a porcine model of septic shock.

Renal cell therapy using the hollow fiber based renal assist device (RAD) improved survival time in an animal model of septic shock (SS) through the amelioration of cardiac and vascular dysfunction. Safety and ability of the RAD to improve clinical outcomes was demonstrated in a Phase II clinical trial, in which patients had high prevalence of sepsis. Even with these promising results, clinical delivery of cell therapy is hampered by manufacturing hurdles, including cell sourcing, large-scale device manufacture, storage and delivery. To address these limitations, the bioartificial renal epithelial cell system (BRECS) was developed. The BRECS contains human renal tubule epithelial cells derived from adult progenitor cells using enhanced propagation techniques. Cells were seeded onto trabeculated disks of niobium-coated carbon, held within cryopreservable, perfusable, injection-moulded polycarbonate housing. The study objective was to evaluate the BRECS in a porcine model of SS to establish conservation of efficacy after necessary cell sourcing and design modifications; a pre-clinical requirement to move back into clinical trials. SS was incited by peritoneal injection of E. coli simultaneous to insertion of BRECS (n=10) or control (n=15), into the ultrafiltrate biofeedback component of an extracorporeal circuit. Comparable to RAD, prolonged survival of the BRECS cohort was conveyed through stabilization of cardiac output and vascular leak. In conclusion, the demonstration of conserved efficacy with BRECS therapy in a porcine SS model represents a crucial step toward returning renal cell therapy to the clinical setting, initially targeting ICU patients with acute kidney injury requiring continuous renal replacement therapy. Copyright © 2014 John Wiley & Sons, Ltd.

Analysis of intraosseous samples in endotoxemic shock--an experimental study in the anaesthetised pig.

Intraosseous (IO) access is used in emergency situations to allow rapid initiation of treatment. IO access is also sometimes used for blood sampling, although data on accuracy of such sampling in critical illness are limited. There is also a potential risk that bone marrow fragments in IO samples may damage laboratory equipment. It is ethically questionable to perform a simultaneous comparison between IO and arterial/venous sampling in critically ill humans. We have, thus, studied the analytical performance of IO sampling in a porcine septic shock model using a cartridge-based analyser. Eight pigs with endotoxin-induced septic shock were sampled hourly for 6 h, and analysed for blood gases, acid base status, haemoglobin, glucose and lactate using point of care instruments. Samples were taken from three IO cannulae (tibia bilaterally, one with infusion, and humerus), one arterial and one venous. An interaction test was used to assess changes in agreement between methods over time. Bland–Altman plots were constructed to study bias between methods. There were, to a varying extent, differences between IO and arterial/venous levels for all studied variables, but agreement did not change significantly during the experiment. A general finding was a large dispersion of differences between methods. IO sample values should be treated with caution in this setting but may add useful information to the clinical picture. The tibia or humerus may be used for sampling. IO infusion decreases agreement, thus sampling during infusion should be avoided.

A new device for continuous assessment of gut perfusion: proof of concept on a porcine model of septic shock.

IntroductionWe evaluate an innovative device consisting of an enteral feeding tube equipped with a photoplethysmography (PPG) sensor in contact with the duodenal mucosa. This study aims to determine if the PPG signal, composed of a continuous (PDC) and a pulsatile part (PAC), is a reliable method to assess gut perfusion in a porcine model of septic shock.MethodFourteen piglets were anesthetized and mechanically ventilated. They were randomly assigned to two groups: the nonseptic (NS) group received an infusion of Ringer’s lactate solution (RL) alone, the septic (S) group received in addition a suspension of live Pseudomonas aeruginosa. Heart rate (HR), pulse oximetry (SpO2), mean arterial pressure (MAP), cardiac index (CI) and serum lactates were recorded and gut microcirculation (GM) was monitored with a laser Doppler probe applied on the duodenal serosa. PDC and PAC were given by the PPG probe inserted in the duodenum. Data was collected every 15 minutes (t0, t15…) during 150 minutes (t150). After administration of the bacteria suspension (t0), resuscitation maneuvers were performed following a defined algorithm. GM PAC, and PDC were expressed as variation from baseline (GMvar, PACvar, PDCvar). Analysis of variance (ANOVA) with repeated measures was performed to compare hemodynamic variables, with Bonferroni correction as post hoc analysis on t0, t60 and t150.ResultsOne piglet was withdrawn from analysis due to a defective probe. S group (six piglets) received resuscitation therapy while NS group (seven piglets) did not. A significant group effect was found for the all parameters except HR. Post hoc analysis found a significant decrease for GM and PAC at t60. The correlation between PAC, PDC and microcirculatory parameters were as follows: rPACvar-GMvar = 0.496, P <0.001, rPDCvar-GMvar = 0.244; P = 0.002. In the septic group, correlations were as follows: rPAC-lactate = -0.772, P <0.001; rPDC-lactate = -0.681, P <0.01). At the onset of shock, a decrease of PAC, PDC and GM occurred before the alteration of MAP.ConclusionsPAC and PDC decreased at the onset of shock and were correlated with GM and lactate. These results confirm that PPG signal reliably reflects the early perfusion alteration of the gut. Further studies should assess the clinical use of this device.

Confocal laser endomicroscopy reliably detects sepsis-related and treatment-associated changes in intestinal mucosal microcirculation

Microcirculatory alterations play a central role in the pathophysiology of sepsis. We investigated probe-based confocal laser endomicroscopy (pCLE) to assess alterations in mucosal microcirculatory perfusion in vivo in a porcine model of septic shock and in patients fulfilling consensus criteria for severe sepsis. Septic shock was induced using a faecal peritonitis model in anaesthetized, mechanically ventilated pigs. Mucosal microcirculation was assessed using pCLE in the stomach, duodenum, terminal ileum, and rectum. Duodenal microcirculation was further evaluated in 10 patients with severe sepsis and in 8 healthy controls to quantify capillary diameter, capillary length, and functional capillary density (FCD). In the animal model, FCD was markedly decreased in duodenal (P<0.001), ileal (P<0.001), gastric (P<0.001), and rectal mucosa (P<0.005) 4 h after induction of sepsis. After volume therapy, FCD partially recovered to 90.0% (duodenum), 94.4% (ileum), 95.4% (gastric), and 97% (rectum) of baseline values, indicating decoupling of microvascular and macrovascular flow. In septic patients, the mean capillary diameter (P<0.01) and FCD (P<0.05) in duodenal mucosa were decreased compared with healthy controls. pCLE reliably detected and quantified microcirculatory alterations in the gastrointestinal mucosa in a porcine model of sepsis and in patients with severe sepsis. Our data suggest that pCLE is a promising tool to assess the efficacy of therapeutic interventions on mucosal microcirculation in real-time, even in the clinical context.

Counteraction of early circulatory derangement by administration of low dose steroid treatment at the onset of established endotoxemic shock is not directly mediated by TNF-α and IL-6

BackgroundOnce a septic condition is progressing, administration of steroids in the pro-inflammatory phase of septic shock ought to yield maximal effect on the subsequent, devastating inflammatory response. Recently, a retrospective study showed that early initiation of corticosteroid therapy improved survival in septic shock. We aimed to prospectively evaluate effects of early administrated hydrocortisone therapy on physiologic variables in a porcine model of septic shock. ExperimentEight anesthetized pigs were given a continuous infusion of endotoxin during this 6h prospective, randomized, parallel-grouped placebo-controlled experimental study. At the onset of endotoxemic shock, defined as the moment when the mean pulmonary arterial pressure reached the double baseline value, the pigs were either given a single intravenous dose of hydrocortisone (5mgkg−1) or the corresponding volume of saline. ResultsMean arterial pressure and systemic vascular resistance index were significantly higher (both p<0.05), and heart rate was significantly lower (p<0.05), in the endotoxin+hydrocortisone group as compared to the endotoxin+saline group. Body temperature and blood hemoglobin levels increased significantly in the endotoxin+saline group (both p<0.05). Urinary hydrocortisone increased significantly in both groups (p<0.05). There were no significant differences in the plasma levels of TNF-alpha, IL-6 or nitrite/nitrate between the groups. ConclusionEarly treatment with hydrocortisone ameliorates some endotoxin mediated circulatory derangements, fever response and microvascular outflow. Our results suggest that these effects are not directly mediated by the pro-inflammatory cytokines TNF-alpha or IL-6, nor by NO.

24 Confocal Laser Endomicroscopy to Assess Mucosal Microcirculation: A Quantitative Analysis in a Porcine Model of Septic Shock and in Patients With Severe Sepsis

Aim Microcirculatory alterations play a central role in the pathophysiology of sepsis. Even if systemic hemodynamic parameters appear to be adequate, septic patients may suffer from significant intestinal microcirculatory alterations. It has been suggested that these changes play a central role in regard to morbidity and mortality. The present study first assessed the feasibility of In Vivo detection of mucosal microcirculation in different segments of the gastrointestinal tract in an animal model of septic shock using probe-based confocal laser endomicroscopy (pCLE) (Mauna Kea Technologies, Paris, France). Subsequently, we assessed duodenal microcirculation in patients suffering from severe sepsis in the early phase of the disease. Materials and Methods First, anesthetized and mechanically ventilated pigs were observed over 8 hrs. Septic shock was triggered by inducing fecal peritonitis (0.75 g autologous feces per kg body weight). Mucosal microcirculation was assessed simultaneously using pCLE in stomach, duodenum, terminal ileum and rectum at baseline, 4 hours after induction of septic shock as well as 2 hours after treatment of the condition. Second, six patients who presented with a severe sepsis on the intensive care unit were examined regarding their duodenal microcirculation after obtaining informed consent from their nearest relatives. These patients were compared to ten healthy controls. Four to six areas were examined for each site and images were analysed in a blinded fashion offline thereafter in both parts of the study. Mean capillary diameter, capillary length and functional capillary density (FCD) were measured quantitatively. Results Two hours after induction of sepsis in the animal model, FCD was markedly decreased in the duodenal (-20.8, p < 0.001), the ileal (-13.4, p < 0.001), the gastric (-11.9%, p < 0.001), and in the rectal mucosal beds (-5.5, p<0,01). After administration of 30 mg x kg-1 of gelatine (30 kDa) FCD increased in all mucosal compartments to 90.0% (duodenum), 94.4% (ileum), 95.4% (gastric) and 97% (rectum) of baseline values. Interestingly, mean vessel diameter was unchanged in all compartments investigated. In patients suffering from severe sepsis we found a significant decrease in mean vessel diameter (-5.0%, p<0.001) as well as in FCD (-9,4%, p<0.001) of the duodenal mucosa, as well. Conclusions During the early phase of septic shock, pCLE may be able to quantify microcirculatory alterations in the gastrointestinal mucosa. Fluid resuscitation improves but does not completely restore intestinal microcirculation in the septic shock model. pCLE may represent a useful tool in order to assess the efficacy of therapeutic interventions on mucosal microcirculation, possibly even in clinical practice.

Model-based cardiovascular monitoring of large pore hemofiltration during endotoxic shock in pigs

The aim of this research is to test the ability of a model-based method to track disease-dependent hemodynamic changes in sepsis. Thus, subject-specific models of the cardiovascular system (CVS) are identified using measurements from a porcine model of septic shock with hemofiltration [1].

LEVOSIMENDAN INCREASES PORTAL BLOOD FLOW AND ATTENUATES INTESTINAL INTRAMUCOSAL ACIDOSIS IN EXPERIMENTAL SEPTIC SHOCK

It has been proposed that vasodilatory therapy may increase microcirculatory blood flow and improve tissue oxygenation in septic shock. The authors aimed to evaluate the effects of levosimendan in systemic and splanchnic hemodynamics in a porcine model of septic shock in a randomized animal controlled study. This study was performed in an animal research facility in a university hospital. Anesthetized pigs were monitored with a pulmonary artery catheter and an ultrasonic blood flow probe in the portal vein for measurement of systemic and portal blood flows and with a tonometer placed in the small intestine for measurement of the intramucosal-arterial PCO2 gap. Three groups of pigs were studied: nonseptic (n = 7), septic (n = 7), and septic treated with levosimendan (n = 7). Levosimendan was administered i.v. at t = -10 min (200 microg/kg in i.v. bolus followed by 200 microg/kg per h). Sepsis was induced at t = 0 min by the administration of live Escherichia coli. Vascular reactivity was tested by the hemodynamic response to noradrenaline. Levosimendan markedly attenuated the sepsis-induced increase in pulmonary vascular resistance, decrease in portal/systemic blood flow, oliguria, impairment in oxygenation, hyperkalemia, and the widened intramucosal-arterial PCO2 gap. Systemic blood pressure and vascular resistance did not differ as compared with the septic untreated group. Responses to noradrenaline significantly improved in animals treated with levosimendan. Treatment with levosimendan in this animal model of sepsis attenuated pulmonary vasoconstriction and improved portal blood flow, intestinal mucosal oxygenation, pulmonary function, and vascular reactivity.

Cardiovascular and Metabolic Effects of High-dose Insulin in a Porcine Septic Shock Model

High-dose insulin (HDI) has inotropic and vasodilatory properties in various clinical conditions associated with myocardial depression. The authors hypothesized that HDI will improve the myocardial depression produced by severe septic shock and have beneficial effects on metabolic parameters. In an animal model of severe septic shock, this study compared the effects of HDI treatment to normal saline (NS) resuscitation alone. Ten pigs were randomized to an insulin (HDI) or NS group. All were anesthetized and instrumented to monitor cardiovascular function. In both arms, Escherichia coli endotoxin lipopolysaccharide (LPS) and NS infusions were begun. LPS was titrated to 20 mug/kg/hour over 30 minutes and continued for 5 hours, and saline was infused at 20 mL/kg/hour throughout the protocol. Dextrose (50%) was infused to maintain glucose in the 60-150 mg/dL range, and potassium was infused to maintain a level greater than 2.8 mmol/L. At 60 minutes, the HDI group received an insulin infusion titrated from 2 to 10 units/kg/hour over 40 minutes and continued at that rate throughout the protocol. Survival, heart rate (HR), mean arterial pressure (MAP), pulmonary artery and central venous pressure, cardiac output, central venous oxygen saturation (SVO(2)), and lactate were monitored for 5 hours (three pigs each arm) or 7 hours (two pigs each arm) or until death. Cardiac index, systemic vascular resistance (SVR), pulmonary vascular resistance (PVR), O(2) delivery, and O(2) consumption were derived from measured data. Outcomes from the repeated-measures analysis were modeled using a mixed-effects linear model that assumed normally distributed errors and a random effect at the subject level. No significant baseline differences existed between arms at time 0 or 60 minutes. Survival was 100% in the HDI arm and 60% in the NS arm. Cardiovascular variables were significantly better in the HDI arm: cardiac index (p < 0.001), SVR (p < 0.003), and PVR (p < 0.01). The metabolic parameters were also significantly better in the HDI arm: SVO(2) (p < 0.01), O(2) delivery (p < 0.001), and O(2) consumption (p < 0.001). No differences in MAP, HR, or lactate were found. In this animal model of endotoxemic-induced septic shock that results in severe myocardial depression, HDI is associated with improved cardiac function compared to NS resuscitation alone. HDI also demonstrated favorable metabolic, pulmonary, and peripheral vascular effects. Further studies may define a potential role for the use of HDI in the resuscitation of septic shock.

COUPLED PLASMA FILTRATION ADSORPTION IN EXPERIMENTAL PERITONITIS-INDUCED SEPTIC SHOCK

The coupled plasma filtration adsorption (CPFA) was developed as an adsorptive hemopurification method aimed at nonselective removal of circulating soluble mediators potentially involved in the pathogenesis of sepsis. We hypothesized that this nonselective hemopurification could protect from detrimental consequences of long-term, volume-resuscitated porcine septic shock. In 16 anesthetized, mechanically ventilated, and instrumented pigs, the hyperdynamic septic shock secondary to peritonitis was induced by intraperitoneally inoculating feces and maintained for 22 h with fluid resuscitation and norepinephrine infusion as needed to maintain MAP above 65 mmHg. After 12 h of peritonitis, animals were randomized to receive either supportive treatment (control, n = 8) or CPFA treatment (CPFA, n = 8). Systemic, hepatosplanchnic, and renal hemodynamics; oxygen exchange; energy metabolism (lactate/pyruvate and ketone body ratios); ileal mucosal and renal cortex microcirculation; systemic inflammation (TNF-alpha, IL-6); nitrosative/oxidative stress (thiobarbituric acid reactive species, nitrates + nitrites); and endothelial/coagulation dysfunction (asymmetric dimethylarginine, von Willebrand factor, thrombin-antithrombin complexes, platelet count) were assessed before and 12, 18, and 22 h of peritonitis. Coupled plasma filtration adsorption neither delayed the development of hypotension nor reduced the dose of norepinephrine. The treatment failed to attenuate sepsis-induced alterations in microcirculation, surrogate markers of cellular energetics, endothelial injury, and systemic inflammation. Similarly, CPFA did not protect from lung and liver dysfunction and even aggravated sepsis-induced disturbances in coagulation and oxidative/nitrosative stress. In this porcine model of septic shock, the early treatment with CPFA was not capable of reversing the sepsis-induced disturbances in various biological pathways and organ systems. Both the efficacy and safety of this method require further rigorous experimental validation in clinically relevant models.

Effects of sodium nitroprusside on splanchnic microcirculation in a resuscitated porcine model of septic shock

We tested the hypothesis that sodium nitroprusside (SNP) might improve the impairment of hepatosplanchnic microcirculatory blood flow (MBF) in septic shock. Fourteen pigs were anaesthetized and their lungs mechanically ventilated. Sepsis was induced with i.v. infusion of live Pseudomonas aeruginosa [1x10(8) colony forming units (CFU) ml(-1) kg(-1)] for 1 h. Sixty minutes later, the animals received in a random succession either SNP or normal saline for 30 min. Mean arterial pressure (MAP), cardiac index (CI), mean pulmonary artery pressure (MPAP), carbon dioxide tension of the ileal mucosa (PCO2; by gas tonometry), ileal mucosal and hepatic MBF by laser Doppler flowmetry, blood gases, and lactates were assessed before, during administration, and 30 min after discontinuing the test drug. Bacterial infusion promoted hypodynamic shock (MAP -18%, CI -33%, ileal MBF -19%, and hepatic MBF -27%), which was converted to normodynamic shock by resuscitation. During SNP infusion, ileal mucosal MBF significantly increased (+19%) compared with control (P = 0.033). Although hepatic MBF increased (+42% from baseline), this did not differ from control. In order to maintain a constant central venous pressure and MAP, fluid loading and norepinephrine (P < 0.01) were increased. Acid-base status was not altered by SNP. In a resuscitated porcine model of the early phase of septic shock, SNP improved ileal mucosal MBF but required a concomitant increase in fluid and norepinephrine supplements to maintain constant systemic haemodynamic parameters.

Vasopressin increases cortisol levels independent of ACTH or catecholamine stimulation in a porcine model of septic shock

Vasopressin increases cortisol levels independent of ACTH or catecholamine stimulation in a porcine model of septic shock

Attenuation of capillary leakage by hydroxyethyl starch (130/0.42) in a porcine model of septic shock

Capillary leakage, a frequent complication in septic shock, is characterized by loss of intravasal fluid resulting in generalized edema and hemodynamic instability despite massive fluid therapy. We have shown that administration of an established colloid 200/0.5 hydroxyethyl starch (HES) stabilized plasma volume in a porcine septic shock model. Recently, a new HES with a low molecular weight (130 kD) and lower molar substitution (0.42) has been developed. In this study, we compared effects of HES 130/0.42 and HES 200/0.5 on capillary leakage in porcine septic shock. Prospective randomized, controlled animal study. University department of anesthesiology. Fourteen pigs (22.9 +/- 2.8 kg). Anesthetized and mechanically ventilated pigs were observed over 6 hrs. Septic shock was induced with fecal peritonitis (0.75 g.kg of body weight autologous feces). Animals were allocated to volume-replacement therapy with either HES 130/0.42 (n = 5) or HES 200/0.5 (n = 5) and compared with nonseptic controls receiving HES 130/0.42 (n = 4). Infusion rate was titrated to maintain a central venous pressure of 12 mm Hg. Albumin escape rate was calculated using iodine 125-labeled albumin. Plasma volume was determined using chromium-51-tagged erythrocytes. Albumin escape rate increased significantly in both groups in comparison to controls (HES 200/0.5, 45% +/- 3; HES 130/0.42, 38% +/- 5), but this increase was significantly smaller with HES 130/0.42. Both HES 200/0.5 (-14%, not significant) and HES 130/0.42 (-1%, not significant) stabilized plasma volume compared with controls. Systemic oxygenation was not significantly altered in either group. In this porcine septic shock model, HES 130/0.42 attenuated capillary leakage significantly more effectively than HES 200/0.5.