Effect of Negative Pressure Therapy on the Inflammatory Response of the Intestinal Microenvironment in a Porcine Septic Model.

In a swine model of ischemia/reperfusion injury coupled with sepsis, we have previously shown attenuation of secondary organ injury and decreased mortality with negative pressure therapy (NPT). We hypothesized that NPT modulates the intestinal microenvironment by mediating the innate immune system. Sepsis was induced in 12 anesthetized female pigs. Group 1 (n = 6) was decompressed at 12 hrs after injury (T 12) and treated with standard of care (SOC), and group 2 (n = 6) with NPT for up to T 48. Immunoparalysis was evident as lymphocytopenia at T 24 in both groups; however, survival was improved in the NPT group versus SOC (Odds ratio = 4.0). The SOC group showed significant reduction in lymphocyte numbers compared to NPT group by T 48 (p < 0.05). The capacity of peritoneal fluid to stimulate a robust reactive oxygen species response in vitro was greater for the NPT group, peaking at T 24 for both M1 (p = 0.0197) and M2 macrophages (p = 0.085). Plasma elicited little if any effect which was confirmed by microarray analysis. In this septic swine model NPT appeared to modulate the intestinal microenvironment, facilitating an early robust, yet transient, host defense mediated by M1 and M2 macrophages. NPT may help overcome immunoparalysis that occurs during inflammatory response to septic injury.

Burst strength testing of porcine intestinal anastomoses following negative pressure therapy.

The effect of negative pressure therapy (NPT; The ABThera™ Open Abdomen Negative Pressure Therapy System, KCI USA, Inc., San Antonio, TX) on the integrity of small intestinal anastomoses was evaluated using in situ burst strength testing in a domestic pig model. In each of 3 swine, 8 anastomoses were created, 4 using sutures and 4 using staples. After 24 hours of continuous NPT, each anastomosis was subjected to burst strength testing in situ. Mean ratios of burst strength of sutured anastomoses to baseline intraluminal pressure were 9.0 to 10.9. Stapled anastomoses had significantly lower burst strength than sutured anastomoses, but mean values were still at least 4.6 times greater than baseline. No differences were seen between anastomoses that were located in close proximity with treatment and those remotely placed or when measured with negative pressure on or off at burst assessment. NPT had no acute adverse effect on intestinal anastomoses in swine.

Peritoneal fluid: a potential mechanism of systemic neutrophil priming in experimental intra-abdominal sepsis.

BACKGROUND: Recent studies suggest that peritoneal fluid (PF) may be an important mediator of inflammation. The aim of this study was to test the hypothesis that PF may drive systemic inflammation in intra-abdominal sepsis by representing a priming agent for neutrophils. METHODS: PF was collected 12 hours after the initiation of intra-abdominal sepsis in swine. Naive human neutrophils were primed with PF before treatment with N-formyl-Met-Leu-Phe or phorbol 12-myristate 13-acetate to elucidate receptor-dependent and receptor-independent mechanisms of neutrophil activation. Flow cytometry was used to quantify neutrophil surface adhesion marker expression of integrins and selectins and superoxide anion production. Additionally, proinflammatory cytokines were quantified in PF. RESULTS: PF primed neutrophils via receptor-dependent and receptor-independent mechanisms. There were significant increases in the proinflammatory cytokines interleukin-6 and tumor necrosis factor-α in PF correlating with the development of intra-abdominal sepsis. CONCLUSIONS: PF represents a priming agent for naive polymorphonuclear cells in intra-abdominal sepsis. This may be secondary to increased levels of proinflammatory cytokines. Strategies to reduce the amount of PF may decrease the systemic inflammatory response by reducing a priming agent for neutrophils.

A novel mechanism for neutrophil priming in trauma: potential role of peritoneal fluid.

BACKGROUND: We sought to determine the effect of peritoneal fluid from a novel animal model of abdominal compartment syndrome (ACS) on the proinflammatory status of polymorphonuclear leukocytes (PMNs) and monocytes. We hypothesize that peritoneal fluid is a potential priming and/or activating agent for PMNs/monocytes. METHODS: ACS was induced in female Yorkshire swine, and peritoneal fluid was collected at the time of decompressive laparotomy. Naïve PMNs/monocytes were primed and/or activated with peritoneal fluid, phosphatidylcholine (PAF) plus peritoneal fluid, peritoneal fluid plus n-formyl-met-leu-phe (fMLP), and peritoneal fluid plus phorbol 12-myristate 13-acetate (PMA). Activation was determined by surface marker expression of integrins (CD11b an CD18) and selectins (CD62L). Additionally, proinflammatory cytokines in peritoneal fluid were analyzed. RESULTS: Peritoneal fluid did not activate PMNs but increased CD11b expression on monocytes. When used as a primer for fMLP- or PMA-induced activation, peritoneal fluid significantly increased CD11b and CD18 expression on PMNs and monocytes. Peritoneal fluid collected at 6 and 12 h post decompressive laparotomy had similar effects. Interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) levels were increased in peritoneal fluid. CONCLUSION: Peritoneal fluid represents a primer for PMNs/monocytes and seems to act through receptor-dependent and receptor-independent pathways. Strategies to reduce the amount of peritoneal fluid may decrease the locoregional and systemic inflammatory response by reducing priming and activation of neutrophils/monocytes.

A novel physiologic model for the study of abdominal compartment syndrome (ACS).

BACKGROUND: : Current abdominal compartment syndrome (ACS) models rely on intraperitoneal instillation of fluid, air, and other space-occupying substances. Although this allows for the study of the effects of increased abdominal pressure, it poorly mimics its pathogenesis. We have developed the first reported large animal model of ACS incorporating hemorrhagic shock/resuscitation. METHODS: : Hemorrhagic shock was induced and maintained (1 hour) in 12 Yorkshire swine by bleeding to a mean arterial pressure (MAP) of 50 mm Hg. The collected blood plus two additional volumes of crystalloid was then reinfused. Mesenteric venous hypertension was induced by tightening a previously placed portal vein snare in a nonocclusive manner to mimic the effects of abdominal packing. Crystalloids were infused to maintain MAP. Hemodynamic measurements, abdominal pressure, peak inspiratory pressures, urine output, and blood chemistries were measured sequentially. Animals were studied for 36 hours after decompression. RESULTS: : ACS (intra-abdominal pressure of > or =20 mm Hg with new organ dysfunction) developed in all animals. There were significant increases in peak inspiratory pressure, central venous pressure, and pulmonary artery pressure and decreases in MAP upon development of ACS. Urine output was significantly decreased before decompression. Mean blood lactate decreased and base excess increased significantly after decompression. CONCLUSIONS: : We have created the first reported physiologic animal ACS model incorporating hemorrhagic shock/resuscitation and the effects of damage control surgery.

Cardiopulmonary effects of continuous negative pressure wound therapy in swine.

BACKGROUND: Negative pressure wound therapy (NPWT) has been used for complex sternotomy wounds. Some reports describe foam placement below the posterior sternal table. We compared the hemodynamic and pulmonary effects of foam location during NPWT after median sternotomy. METHODS: Swine were randomized into four groups (n = 6 per group). A polyurethane open cell foam dressing was placed either within or below the sternal table. In one-half, a silicone mesh barrier was placed between the heart and the foam. The NPWT was applied at -125 mm Hg and then released to ambient pressure. This cycle was repeated two more times, and the foam was removed. Heart rate, mean arterial pressure, cardiac output, mixed venous oxygenation, central venous pressure, and pulmonary artery wedge pressure were measured. Peak inspiratory pressure, mean airway pressure, work of breathing, and intrathoracic pressure measurements were recorded. RESULTS: Intersternal placement of foam did not affect hemodynamic parameters. Substernal placement resulted in depression of hemodynamic variables which improved when negative pressure was applied. Pulmonary mechanics were not affected by foam location. CONCLUSIONS: Initial placement of the foam dressing below the posterior sternal table caused reversible depression of cardiac function which appears to be consistent with direct cardiac compression. NPWT therapy had no clinically significant impact on pulmonary parameters. The use of a protective barrier does not alter hemodynamic or pulmonary parameters but continues to be recommended when NPWT is used for sternotomy wounds.