Influence of endothelial glycocalyx degradation and surfactants on air embolism adhesion

BACKGROUND: Microbubble adherence to endothelial cells is enhanced after damage to the glycocalyx. The authors tested the hypothesis that exogenous surfactants delivered intravascularly have differential effects on the rate of restoration of blood flow after heparinase-induced degradation of the endothelial glycocalyx. METHODS: Air microbubbles were injected into the rat cremaster microcirculation after perfusion with heparinase or saline and intravascular administration of either saline or one of two surfactants. The surfactants were Pluronic F-127 (Molecular Probes, Eugene, OR) and Perftoran (OJSC SPC Perftoran, Moscow, Russia). Embolism dimensions and dynamics were observed using intravital microscopy. RESULTS: Significant results were that bubbles embolized the largest diameter vessels after glycocalyx degradation. Bubbles embolized smaller vessels in the surfactant treatment groups. The incidence of bubble dislodgement and the magnitude of distal displacement were smallest after glycocalyx degradation alone and largest after surfactant alone. The time to bubble clearance and restoration of blood flow was longest with heparinase alone and shortest with Pluronic F-127 alone. CONCLUSIONS: Degradation of the glycocalyx causes air bubbles to adhere to the endothelium more proximally in the arteriolar microcirculation. Surfactants added after glycocalyx degradation and before gas embolization promotes bubble lodging in the distal microcirculation. Surfactants may have a clinical role in reducing embolism bubble adhesion to endothelial cells undergoing glycocalyx disruption.     

Surfactant Reduction in Embolism Bubble Adhesion and Endothelial Damage

Background: Surfactants may reduce the adhesion force holding bubbles to the vessel wall in gas embolism. The authors measured bubble adhesion force using excised microvessels. They assessed endothelial damage by measuring vessel reactivity and with microscopy.

Methods: Microbubbles injected into arterioles resided for 5, 10, or 30 min, with intact or damaged endothelium. Perfusion was with rat serum alone (control) or with 1% Perftoran (OJSC SPC Perftoran, Moscow, Russia) or 1% Pluronic F-127 (Molecular Probes, Eugene, OR) added. Pressure across the bubble, bubble length, and bubble diameter were measured, and adhesion force per unit surface area, K = [DELTA]PD/4 l, was calculated. Vessel reactivity was assessed using topical application of phenylephrine and acetylcholine.

Results: With the endothelium intact, K was higher in controls than with Perftoran at 10 and 30 min or Pluronic F-127 at 10 min (P < 0.05). With surfactant added after air perfusion to damage the endothelium, K was lower (P < 0.05) at all times for both Perftoran and Pluronic F-127. With surfactant in the perfusate before air perfusion, K was lower at 10 and 30 min for Perftoran and at 10 min for Pluronic F-127 than for controls (P < 0.05). Phenylephrine-induced vasoconstriction was identical among groups. Acetylcholine-induced vasodilatation was the same among groups with an intact endothelium but was found to be lower in controls after air perfusion that followed surfactant exposure than in either surfactant group (P < 0.05).     

Surfactant reduction in embolism bubble adhesion and endothelial damage

BACKGROUND: Surfactants may reduce the adhesion force holding bubbles to the vessel wall in gas embolism. The authors measured bubble adhesion force using excised microvessels. They assessed endothelial damage by measuring vessel reactivity and with microscopy. METHODS: Microbubbles injected into arterioles resided for 5, 10, or 30 min, with intact or damaged endothelium. Perfusion was with rat serum alone (control) or with 1% Perftoran (OJSC SPC Perftoran, Moscow, Russia) or 1% Pluronic F-127 (Molecular Probes, Eugene, OR) added. Pressure across the bubble, bubble length, and bubble diameter were measured, and adhesion force per unit surface area, K = deltaPD/4 l, was calculated. Vessel reactivity was assessed using topical application of phenylephrine and acetylcholine. RESULTS: With the endothelium intact, K was higher in controls than with Perftoran at 10 and 30 min or Pluronic F-127 at 10 min (P < 0.05). With surfactant added after air perfusion to damage the endothelium, K was lower (P < 0.05) at all times for both Perftoran and Pluronic F-127. With surfactant in the perfusate before air perfusion, K was lower at 10 and 30 min for Perftoran and at 10 min for Pluronic F-127 than for controls (P < 0.05). Phenylephrine-induced vasoconstriction was identical among groups. Acetylcholine-induced vasodilatation was the same among groups with an intact endothelium but was found to be lower in controls after air perfusion that followed surfactant exposure than in either surfactant group (P < 0.05). CONCLUSIONS: Surfactants reduced bubble adhesion force and preserved basic endothelial structure and vasodilatory function despite attempts to damage the endothelium. Surfactants seem to protect the endothelium from mechanically induced injury in addition to decreasing bubble adhesion forces.     

Surfactants Reduce Platelet-Bubble and Platelet-Platelet Binding Induced by In Vitro Air Embolism

Background: The effect of gas bubbles on platelet behavior is poorly characterized. The authors assessed platelet-bubble and platelet-platelet binding in platelet-rich plasma in the presence and absence of bubbles and three surface-active compounds.

Methods: Platelet-rich plasma was prepared from blood drawn from 16 volunteers. Experimental groups were surfactant alone, sparging (microbubble embolization) alone, sparging with surfactant, and neither sparging nor surfactant. The surfactants were Pluronic F-127 (Molecular Probes, Eugene, OR), Perftoran (OJSC SPC Perftoran, Moscow, Russia), and Dow Corning Antifoam 1510US (Dow Corning, Midland, MI). Videomicroscopy images of specimens drawn through rectangular glass microcapillaries on an inverted microscope and Coulter counter measurements were used to assess platelet-bubble and platelet-platelet binding, respectively, in calcium-free and recalcified samples. Histamine-induced and adenosine diphosphate-induced platelet-platelet binding were measured in unsparged samples. Differences between groups were considered significant for P < 0.05 using analysis of variance and the Bonferroni correction.

Results: Sixty to 100 platelets adhered to bubbles in sparged, surfactant-free samples. With sparging and surfactant, few platelets adhered to bubbles. Numbers of platelet singlets and multimers not adherent to bubbles were different (P < 0.05) compared both with unsparged samples and sparged samples without surfactant. No significant platelet-platelet binding occurred in uncalcified, sparged samples, although 20-30 platelets adhered to bubbles. Without sparging, histamine and adenosine diphosphate provoked platelet-platelet binding with and without surfactants present.     

Surfactants Attenuate Gas Embolism-induced Thrombin Production

Background: There are no pharmacologic strategies to prevent embolism bubble-induced blood clot formation. The authors conducted experiments to measure thrombin production in sheared whole blood in the presence and absence of bubbles and three surface-active compounds.

Methods: Blood samples were obtained from six volunteers seven times. The thrombin-specific substrate Boc-VPR-MCA was added to citrated blood diluted with HEPES-buffered saline. Experimental groups were as follows: sparging (air microbubble embolization) with surfactant present; sparging alone; surfactant alone; and neither surfactant nor sparging. The surfactants were Dow Corning Antifoam 1510US, Perftoran, and Pluronic F-127. Blood was sheared by a cone-plate viscometer at 100 and 500 s-1 for 5, 10, and 20 min at 37[degrees]C, pipetted into excess stop buffer, and evaluated fluorimetrically. Mean values of fluorescence intensity +/- SDs for each group were compared using ANOVA. Differences were considered significant at P < 0.05 using the Bonferroni correction.

Results: For fixed shear rate, thrombin production increased 2.3- to 5.7-fold (P < 0.05) as shear duration lengthened. For fixed shear duration, thrombin production increased 1.9- to 3.9-fold (P < 0.05) with increasing shear rate. For fixed shear rate and duration, sparging increased thrombin production 2.1- to 3.7-fold (P < 0.05). Surfactant addition without sparging did not change thrombin production (P > 0.05). Surfactants attenuated thrombin production in sparged samples 31.8-70.9% (P < 0.05).     

Endothelial Glycocalyx as an Additional Barrier Determining Extravasation of 6% Hydroxyethyl Starch or 5% Albumin Solutions in the Coronary Vascular Bed

Background: The impact on the endothelial glycocalyx for the extravasation of colloidal infusion solutions has not been investigated sufficiently.

Methods: Isolated guinea pig hearts were perfused with Krebs-Henseleit buffer in a Langendorff mode. Solutions of 0.9% saline, 5% albumin (70 kd), or 6% hydroxyethyl starch (200 kd) were infused into the coronary system for 20 min at a rate of one third of the coronary flow, also during reperfusion after 15 min of ischemia, and after enzymatic digestion of the endothelial glycocalyx by heparinase. Net coronary fluid filtration was assessed directly by measuring the formation of transudate on the epicardial surface, and solute extravasation was assessed by measuring albumin and hydroxyethyl starch in the coronary effluent and transudate. Hearts were perfusion fixed to visualize the endothelial glycocalyx using transmission electron microscopy.

Results: Only infusion of hydroxyethyl starch, not infusion of albumin, significantly decreased net coronary fluid filtration. Heparinase application without ischemia increased coronary leak by 25% but did not accelerate the passage of colloids. Ischemia alone did not alter permeability. However, there was a large (approximately +200%), transient (approximately 4 min) increase in permeability for water, albumin, and hydroxyethyl starch after ischemia with heparinase application. Also, histamine (10-6m) only increased permeability after pretreatment of the hearts with heparinase. The thickness of the glycocalyx after colloid administration was 0.2-0.3 [mu]m. No glycocalyx could be detected after application of heparinase.     

Surfactants reduce platelet-bubble and platelet-platelet binding induced by in vitro air embolism

BACKGROUND: The effect of gas bubbles on platelet behavior is poorly characterized. The authors assessed platelet-bubble and platelet-platelet binding in platelet-rich plasma in the presence and absence of bubbles and three surface-active compounds. METHODS: Platelet-rich plasma was prepared from blood drawn from 16 volunteers. Experimental groups were surfactant alone, sparging (microbubble embolization) alone, sparging with surfactant, and neither sparging nor surfactant. The surfactants were Pluronic F-127 (Molecular Probes, Eugene, OR), Perftoran (OJSC SPC Perftoran, Moscow, Russia), and Dow Corning Antifoam 1510US (Dow Corning, Midland, MI). Videomicroscopy images of specimens drawn through rectangular glass microcapillaries on an inverted microscope and Coulter counter measurements were used to assess platelet-bubble and platelet-platelet binding, respectively, in calcium-free and recalcified samples. Histamine-induced and adenosine diphosphate-induced platelet-platelet binding were measured in unsparged samples. Differences between groups were considered significant for P < 0.05 using analysis of variance and the Bonferroni correction. RESULTS: Sixty to 100 platelets adhered to bubbles in sparged, surfactant-free samples. With sparging and surfactant, few platelets adhered to bubbles. Numbers of platelet singlets and multimers not adherent to bubbles were different (P < 0.05) compared both with unsparged samples and sparged samples without surfactant. No significant platelet-platelet binding occurred in uncalcified, sparged samples, although 20-30 platelets adhered to bubbles. Without sparging, histamine and adenosine diphosphate provoked platelet-platelet binding with and without surfactants present. CONCLUSIONS: Sparging causes platelets to bind to air bubbles and each other. Surfactants added before sparging attenuate platelet-bubble and platelet-platelet binding. Surfactants may have a clinical role in attenuating gas embolism-induced platelet-bubble and platelet-platelet binding.     

Surfactants attenuate gas embolism-induced thrombin production

BACKGROUND: There are no pharmacologic strategies to prevent embolism bubble-induced blood clot formation. The authors conducted experiments to measure thrombin production in sheared whole blood in the presence and absence of bubbles and three surface-active compounds. METHODS: Blood samples were obtained from six volunteers seven times. The thrombin-specific substrate Boc-VPR-MCA was added to citrated blood diluted with HEPES-buffered saline. Experimental groups were as follows: sparging (air microbubble embolization) with surfactant present; sparging alone; surfactant alone; and neither surfactant nor sparging. The surfactants were Dow Corning Antifoam 1510US, Perftoran, and Pluronic F-127. Blood was sheared by a cone-plate viscometer at 100 and 500 s-1 for 5, 10, and 20 min at 37 degrees C, pipetted into excess stop buffer, and evaluated fluorimetrically. Mean values of fluorescence intensity +/- SDs for each group were compared using ANOVA. Differences were considered significant at P < 0.05 using the Bonferroni correction. RESULTS: For fixed shear rate, thrombin production increased 2.3- to 5.7-fold (P < 0.05) as shear duration lengthened. For fixed shear duration, thrombin production increased 1.9- to 3.9-fold (P < 0.05) with increasing shear rate. For fixed shear rate and duration, sparging increased thrombin production 2.1- to 3.7-fold (P < 0.05). Surfactant addition without sparging did not change thrombin production (P > 0.05). Surfactants attenuated thrombin production in sparged samples 31.8-70.9% (P < 0.05). CONCLUSIONS: Thrombin production is shear rate and duration-dependent. Sparging increases thrombin production. Surfactants added before sparging attenuate thrombin production. Surfactants may have a clinical application to attenuate gas embolism-induced clotting.     

Endothelial glycocalyx as an additional barrier determining extravasation of 6% hydroxyethyl starch or 5% albumin solutions in the coronary vascular bed

BACKGROUND: The impact on the endothelial glycocalyx for the extravasation of colloidal infusion solutions has not been investigated sufficiently. METHODS: Isolated guinea pig hearts were perfused with Krebs-Henseleit buffer in a Langendorff mode. Solutions of 0.9% saline, 5% albumin (70 kd), or 6% hydroxyethyl starch (200 kd) were infused into the coronary system for 20 min at a rate of one third of the coronary flow, also during reperfusion after 15 min of ischemia, and after enzymatic digestion of the endothelial glycocalyx by heparinase. Net coronary fluid filtration was assessed directly by measuring the formation of transudate on the epicardial surface, and solute extravasation was assessed by measuring albumin and hydroxyethyl starch in the coronary effluent and transudate. Hearts were perfusion fixed to visualize the endothelial glycocalyx using transmission electron microscopy. RESULTS: Only infusion of hydroxyethyl starch, not infusion of albumin, significantly decreased net coronary fluid filtration. Heparinase application without ischemia increased coronary leak by 25% but did not accelerate the passage of colloids. Ischemia alone did not alter permeability. However, there was a large (approximately +200%), transient (approximately 4 min) increase in permeability for water, albumin, and hydroxyethyl starch after ischemia with heparinase application. Also, histamine (10 m) only increased permeability after pretreatment of the hearts with heparinase. The thickness of the glycocalyx after colloid administration was 0.2-0.3 microm. No glycocalyx could be detected after application of heparinase. CONCLUSION: The endothelial glycocalyx acts as a competent barrier for water and colloids. Only after its destruction do changes in endothelial morphology (postischemic reperfusion or histamine application) become effective determinants of coronary extravasation.     

同种异体软骨细胞移植术后关节软骨蛋白多糖的测定

目的 应用Pluronic F-127负载同种异体软骨细胞移植修复兔全厚关节软骨损伤,对于新生的修复组织进行基质蛋白多糖含量测定,以探讨此方法修复全厚关节软骨损伤的可行性.方法 取3个月龄新西兰大白兔关节软骨细胞体外培养扩增,与20%Plurortic F-127凝胶混合.选27只健康同种成年大白兔,人为造成双侧膝关节软骨缺损.实验组软骨缺损处植入培养的软骨细胞/Pluronic F-127混合物,对照组缺损处单纯注入Pluronic F-127凝胶和空白对照.然后,对修复组织进行大体观察及蛋白多糖含量测定.结果 移植的软骨细胞-载体复合物中的软骨细胞能良好地生长,12周时再生组织与周围正常软骨组织外观相似,界限模糊.实验组与对照组各时期蛋白多糖含量均有非常显著性差异,实验组不同时期的蛋白多糖含量之间均有显著性差异,实验组12周时蛋白多糖含量与正常软骨组织无显著性差异.结论 Pluronic F-127负载同种异体软骨细胞移植是治疗关节软骨缺损的有效方法.     

Contrasting Effects of Colloid and Crystalloid Resuscitation Fluids on Cardiac Vascular Permeability

Background: Fluid extravasation may lead to myocardial edema and consequent reduction in ventricular function. Albumin is presumed to interact with the endothelial glycocalyx. The authors' objective was to compare the impact of different resuscitation fluids (human albumin, hydroxyethyl starch, saline) on vascular integrity.

Methods: In an isolated perfused heart model (guinea pig), Krebs-Henseleit buffer was augmented with colloids (one third volume 5% albumin or 6% hydroxyethyl starch 130/0.4) or crystalloid (0.9% saline). Perfusion pressure and vascular fluid filtration (epicardial transudate formation) were assessed at different flow rates. After global, stopped-flow ischemia (37[degrees]C, 20 min), hearts were reperfused with the same resuscitation fluid additives. In a second series, the authors applied the respective perfusates after enzymatic digestion of the endothelial glycocalyx (heparinase, 10 U over 15 min).

Results: Both 5% albumin and 6% hydroxyethyl starch decreased fluid extravasation versus saline (68.4 +/- 5.9, 134.8 +/- 20.5, and 436.8 +/- 14.7 [mu]l/min, respectively, at 60 cm H2O perfusion pressure; P < 0.05), the corresponding colloid osmotic pressures being 2.95, 5.45, and 0.00 mmHg. Digestion of the endothelial glycocalyx decreased coronary integrity in both colloid groups. After ischemia, a transient increase in vascular leak occurred with Krebs-Henseleit buffer containing hydroxyethyl starch and saline, but not with albumin. The authors observed no difference between intravascular and bulk interstitial colloid concentration in the steady state. Notwithstanding, electron microscopy revealed an intact endothelial glycocalyx and no interstitial edema in the albumin group.     

Impact of heparanase on glomerular endothelium glycocalyx during sepsis

Objective To observe the impact of heparanase on glomerular endothelium glycocalyx during sepsis and to investigate the prevention of glycocalyx injury. Methods C57/BL6 mice were injected with lipopolysaccharide (LPS) or tumor necrosis factor-α(TNF-α) and sacrificed one hour later. Glomerular endothelium glycocalyx traced with lanthanum was observed by transmission electron microscope(TEM). Western blotting was used to observe heparanse protein expression of renal cortex tissue. Human renal glomerular endothelial cells (HRGECs) were stimulated with TNF-α and active heparanase protien expression was detected by Western blotting. Mice were administrated with heparin sodium or heparinase Ⅲ and renal endothelium glycocalyx was observed by TEM. Urine during twenty-four hours was collected to measure urinary albumin and creatinine. The ratio of albumin to creatinine was calculated and compared among groups. Results The glomerular endothelium glycocalyx of LPS group and TNF-α group was degradated and the one of podocyte was integrated. Renal cortex tissue heparanase protein expression was significantly increased since one hour after LPS injection (P＜0.01). The protein expression of activited heparanase of HRGECs which were stimulated with TNF-α was increased (P＜0.05). Administration of heparin sodium which could inhibit the activity of heparanase could prevent the glycocalyx form degradation. The ratio of urine albumin to creatinine of heparin sodium group was decreased compared with LPS group (P＜0.05) and the ratio of heparinase Ⅲ group was higher than control group(P＜0.01) as a result of degradation of glomerular endothelium glycocalyx. Conclusions During the early stage of sepsis, TNF-α can induce glomerular endothelium heparanase to increase and active, and consequently the glycocalyx is degradated which leads to albuminuria. Inhibition of heparanase can protect glomerular endothelium glycocalyx and prevent albuminuria.     

Hydrocortisone Preserves the Vascular Barrier by Protecting the Endothelial Glycocalyx

Background: Hydrocortisone protects against ischemia-reperfusion injury, reduces paracellular permeability for macromolecules, and is routinely applied in the prevention of interstitial edema. Healthy vascular endothelium is coated by the endothelial glycocalyx, diminution of which increases capillary permeability, suggesting that the glycocalyx is a target for hydrocortisone action.

Methods: Isolated guinea pig hearts were perfused with Krebs-Henseleit buffer. Hydrocortisone was applied in a stress dose (10 [mu]g/ml) before inducing 20 min of ischemia (37[degrees]C). Hearts were reperfused for 20 min at constant flow (baseline perfusion pressure, 70 cm H2O) with Krebs-Henseleit buffer or Krebs-Henseleit buffer plus 2 g% hydroxyethyl starch (130 kd). Coronary net fluid filtration was assessed directly by measuring transudate formation on the epicardial surface. Hearts were perfusion fixed to visualize the glycocalyx.

Results: Ischemia-induced degradation of the glycocalyx enhanced coronary perfusion pressure (118.8 +/- 17.3 cm H2O) and increased vascular permeability (8 +/- 0.2 [mu]l [middle dot] min-1 [middle dot] cm H2O-1 at baseline vs. 34 +/- 3.3 [mu]l [middle dot] min-1 [middle dot] cm H2O-1 after reperfusion). Enzymatic digestion of the glycocalyx (heparinase) elicited similar effects. Hydrocortisone reduced postischemic oxidative stress, perfusion pressure (86.3 +/- 6.4 cm H2O), and transudate formation (11 +/- 0.6 [mu]l [middle dot] min-1 [middle dot] cm H2O-1). Applying colloid augmented this (70.6 +/- 5.6 cm H2O and 9 +/- 0.5 [mu]l [middle dot] min-1 [middle dot] cm H2O-1). Postischemic shedding of syndecan-1, heparan sulfate, and hyaluronan was inhibited by hydrocortisone, as was release of histamine from resident mast cells. Electron microscopy revealed a mostly intact glycocalyx after hydrocortisone treatment, but not after heparinase treatment.     

Contrasting effects of colloid and crystalloid resuscitation fluids on cardiac vascular permeability

BACKGROUND: Fluid extravasation may lead to myocardial edema and consequent reduction in ventricular function. Albumin is presumed to interact with the endothelial glycocalyx. The authors' objective was to compare the impact of different resuscitation fluids (human albumin, hydroxyethyl starch, saline) on vascular integrity. METHODS: In an isolated perfused heart model (guinea pig), Krebs-Henseleit buffer was augmented with colloids (one third volume 5% albumin or 6% hydroxyethyl starch 130/0.4) or crystalloid (0.9% saline). Perfusion pressure and vascular fluid filtration (epicardial transudate formation) were assessed at different flow rates. After global, stopped-flow ischemia (37 degrees C, 20 min), hearts were reperfused with the same resuscitation fluid additives. In a second series, the authors applied the respective perfusates after enzymatic digestion of the endothelial glycocalyx (heparinase, 10 U over 15 min). RESULTS: Both 5% albumin and 6% hydroxyethyl starch decreased fluid extravasation versus saline (68.4 +/- 5.9, 134.8 +/- 20.5, and 436.8 +/- 14.7 microl/min, respectively, at 60 cm H(2)O perfusion pressure; P < 0.05), the corresponding colloid osmotic pressures being 2.95, 5.45, and 0.00 mmHg. Digestion of the endothelial glycocalyx decreased coronary integrity in both colloid groups. After ischemia, a transient increase in vascular leak occurred with Krebs-Henseleit buffer containing hydroxyethyl starch and saline, but not with albumin. The authors observed no difference between intravascular and bulk interstitial colloid concentration in the steady state. Notwithstanding, electron microscopy revealed an intact endothelial glycocalyx and no interstitial edema in the albumin group. CONCLUSION: Ex vivo, albumin more effectively prevented fluid extravasation in the heart than crystalloid or artificial colloid. This effect was partly independent of colloid osmotic pressure and may be attributable to an interaction of albumin with the endothelial glycocalyx.     

Hydrocortisone preserves the vascular barrier by protecting the endothelial glycocalyx

BACKGROUND: Hydrocortisone protects against ischemia-reperfusion injury, reduces paracellular permeability for macromolecules, and is routinely applied in the prevention of interstitial edema. Healthy vascular endothelium is coated by the endothelial glycocalyx, diminution of which increases capillary permeability, suggesting that the glycocalyx is a target for hydrocortisone action. METHODS: Isolated guinea pig hearts were perfused with Krebs-Henseleit buffer. Hydrocortisone was applied in a stress dose (10 microg/ml) before inducing 20 min of ischemia (37 degrees C). Hearts were reperfused for 20 min at constant flow (baseline perfusion pressure, 70 cm H2O) with Krebs-Henseleit buffer or Krebs-Henseleit buffer plus 2 g% hydroxyethyl starch (130 kd). Coronary net fluid filtration was assessed directly by measuring transudate formation on the epicardial surface. Hearts were perfusion fixed to visualize the glycocalyx. RESULTS: Ischemia-induced degradation of the glycocalyx enhanced coronary perfusion pressure (118.8 +/- 17.3 cm H2O) and increased vascular permeability (8 +/- 0.2 microl x min(-1) x cm H2O(-1) at baseline vs. 34 +/- 3.3 microl x min(-1) x cm H2O(-1) after reperfusion). Enzymatic digestion of the glycocalyx (heparinase) elicited similar effects. Hydrocortisone reduced postischemic oxidative stress, perfusion pressure (86.3 +/- 6.4 cm H2O), and transudate formation (11 +/- 0.6 microl x min(-1) x cm H2O(-1)). Applying colloid augmented this (70.6 +/- 5.6 cm H2O and 9 +/- 0.5 microl x min(-1) x cm H2O(-1)). Postischemic shedding of syndecan-1, heparan sulfate, and hyaluronan was inhibited by hydrocortisone, as was release of histamine from resident mast cells. Electron microscopy revealed a mostly intact glycocalyx after hydrocortisone treatment, but not after heparinase treatment. CONCLUSIONS: Hydrocortisone preserves the endothelial glycocalyx, sustaining the vascular barrier and reducing interstitial edema. The effect of colloids suggests that prevention of postischemic rise in coronary resistance by hydrocortisone could also be based on alleviation of endothelial swelling. Stabilization of myocardial mast cells by hydrocortisone may account for the mitigated inflammatory affect of ischemia-reperfusion.     

The protective role of estrogen on endothelial and glycocalyx barriers after shock conditions: A microfluidic study

BackgroundSexual dimorphism has been demonstrated after major trauma and hemorrhage shock with protective effects related to female sex or estrogen. Traumatic endotheliopathy is an important component of trauma-induced coagulopathy. Components of endothelial barrier dysfunction include degradation of the endothelial glycocalyx and endothelial cellular injury. Estrogen modulates endothelial function via its membrane and cellular receptors. The effects of estrogen on the vascular endothelial barrier after trauma and hemorrhage shock are, however, unknown. This topic was studied in an in vitro model under flow conditions.MethodsMonolayers of human umbilical vein endothelial cells were established in microfluidic flow devices. After overnight perfusion, cell monolayers were subjected to normoxic or hypoxic perfusion and then treated with either estrogen (as estradiol), testosterone (as dihydrotestosterone), or media alone. Endothelial activation/injury was indexed by soluble thrombomodulin and glycocalyx degradation by syndecan-1 and hyaluronic acid shedding as well as measurement of the thickness of the glycocalyx layer. The coagulation phenotype of the human umbilical vein endothelial cells was indexed by the relative values of the activities of tissue plasminogen activator and plasminogen activator inhibitor-1. Vascular endothelial growth factor was measured in cell culture supernatants using a solid-phase enzyme-linked immunosorbent assay.ResultsTreatment with estrogen but not testosterone mitigated the adverse effect of shock on endothelial and glycocalyx barrier properties. Our biomimetic model suggests a beneficial effect of estrogen administration after trauma and hemorrhage shock on the glycocalyx and endothelial barriers.ConclusionEarly estrogen treatment after trauma and hemorrhage shock may be a useful adjunct to mitigating the development of traumatic endotheliopathy.     

啮齿类动物出血性休克模型中血浆对内皮细胞多糖包被的恢复作用

背景在出血性休克的创伤患者应用血浆为主的复苏与死亡率下降有关。尽管有学者提出通过替代凝血蛋白可产生有益的效果,但是输入血浆的保护作用可能机制尚不明了。我们以前在细胞培养模型中的研究显示,与晶体液相比,血浆可降低内皮细胞的通透性。多糖包被由粘附于多配体聚糖骨架上的蛋白多糖和糖蛋白组成,共同保护下层的内皮细胞。我们假设,血浆对内皮细胞的保护作用,部分是由于出血性休克后内皮细胞多糖包被的恢复和多配体聚糖-1的维持所致。方法在大鼠建立出血性休克模型使其平均动脉压在30mmHg维持90分钟,然后以乳酸林格（1actatedRinger＇s,LR）液或新鲜血浆复苏,使其平均动脉压达到80mmHg,相当于假手术组或单纯休克组动物。2小时后,提取肺组织多配体聚糖的mRNA,使用抗多配体聚糖-l免疫染色或使用苏木精伊红染色。为了特异地检测出血浆对内皮细胞的影响,我们通过小肠系膜灌注镧溶液,然后应用电子显微镜鉴别出小静脉,观察其多糖包被。所有数据以均数±标准差来表示。运用单尾方差分析合并Tukey事后检验来分析结果。结果电子显微镜显示出血性休克后多糖包被发生降解,输注血浆可部分恢复多糖包被水平而LR无该效应。血浆复苏组动物肺组织多配体聚糖．1的mRNA表达（2．76±0．03）明显高于单纯休克组（1．39±0．22）或ut组（0．82±0．03）,并且与细胞表面多配体聚糖．1的免疫染色相关。休克也可导致组织病理学评分方面肺损伤显著（1．63±0．26）,应用血浆复苏可减轻该损伤（0．67±0．17）,而LR无该效应（2．0±0．25）。结论血浆的保护作用可能部分是由于出血性休克后血浆能恢复内皮细胞多糖包被水平,并维持多配体聚糖-1水平。     

Mechanical cleansing of contaminated wounds with a surfactant

Mechanical cleansing of a wound with a sponge soaked in a surfactant has prevented the development of experimental wound infection. The surfactant utilized for wound cleansing is Pluronic F-68, a member of a family of block copolymers called Pluronic polyols. Long-term toxicity studies and clinical trials suggest that this surfactant is safe for human use. Pluronic F-68 is a nonionic detergent that does not have any intrinsic antibacterial activity.Although mechanical cleansing with salinesoaked sponges effectively removes bacteria, it damages the wound and impairs its resistance to infection. The severity of the damage to the skin exerted by the sponge can be correlated with its porosity. Sponges with a low porosity are abrasive and exert more damage to skin than do sponges with a higher porosity. The addition of Pluronic F-68 to even the most abrasive sponges ensures that the bacterial removal efficiency of the sponge scrub is maintained, while tissue trauma is minimized. This dual effect of the surfactant results in a dramatic reduction in the infection rate of contaminated wounds. On the basis of these results, a clinical trial with surfactant-soaked sponges would appear to be indicated.     

The molecular basis for toxicity of surfactants in surgical wounds. 1. EO:PO block polymers

The purpose of this study was to ascertain how molecular variations in a homologous series of polymeric nonionic surfactants affect their tissue toxicity. The results of these structure-toxicity studies serve to identify nontoxic surfactants for use in surgical scrub solutions. Structure-toxicity studies were undertaken with a family of nonionic surfactants, Pluronic polyols, whose composition can be easily altered to form an almost unlimited number of nonionic surfactants. Pluronic polyols are a series of block copolymers that consist of water-soluble poly (oxyethylene) groups at both ends of a water-insoluble poly (oxypropylene) chain. Pluronic polyols of varying molecular weight with a wide range of ethylene oxide content were used for this study. The effect of topical application of solutions of Pluronic polyols on the wound's resistance to infection was used as a measure of toxicity.The molecular weight of the Pluronic polyol was found to be an unimportant determinant of the toxicity of these surfactants in the surgical wounds. The incidence of gross infection in contaminated wounds after the application of a polyol with a high molecular weight differed insignificantly from the infection rate of wounds treated with a polyol with a lower molecular weight. The ethylene oxide content in the Pluronic polyol was an important causal factor of toxicity. Contaminated wounds treated with a polyol with a high ethylene oxide content (80% by weight of total molecule) exhibited a lower incidence of infection than wounds receiving a polyol with a lower ethylene oxide content (20–50% by weight of total molecule). Surfactants with a high ethylene oxide content failed to impair the wound's resistance to infection. The gross infection scores for contaminated wounds treated with the polyols with a high ethylene oxide content differed insignificantly from the incidence of infection of wounds subjected to 0.9% sodium chloride.The use of a nontoxic surfactant has improved the therapeutic value of surgical scrub solutions in the treatment of the contaminated wound. A polyol with a high ethylene oxide content, Pluronic F68, has been utilized with an iodophor, polyvinylpyrrolidone-iodine, in the preparation of a new surgical scrub solution. The therapeutic value of this solution was compared to another surgical scrub solution containing polyvinylpyrrolidone-iodine and an anionic surfactant. The incidence of gross infection after treatment with the mixture of the Pluronic polyol and the iodophor was significantly less than the infection rates of wounds subjected to a surgical scrub solution containing an iodophor and an anionic surfactant. The results of the study provide compelling reasons for further experimental and clinical investigations to assess the potential of this nontoxic surfactant in the treatment of the contaminated wound.     

Accelerated Arteriolar Gas Embolism Reabsorption by an Exogenous Surfactant

Background: Cerebrovascular gas embolism can cause profound neurologic dysfunction, and there are few treatments. The authors tested the hypothesis that an exogenous surfactant can be delivered into the bloodstream to alter the air-blood interfacial mechanics of an intravascular gas embolism and produce bubble conformations, which favor more rapid bubble absorption.

Methods: Microbubbles of air were injected into the rat cremaster microcirculation after intravascular administration of either saline (control, n = 5) or Dow Corning Antifoam 1510US (surfactant, n = 5). Embolism dimensions and dynamics were directly observed after entrapment using intravital microscopy.

Results: To achieve embolization, the surfactant group required twice as many injections as did controls (3.2 +/- 1.3 vs. 1.6 +/- 0.9;P < 0.05). There was no difference in the initial lodging configuration between groups. After bubble entrapment, there was significantly more local vasoconstriction in the surfactant group (24.2% average decrease in diameter) than in controls (3.4%;P < 0.05). This was accompanied by a 92.7% bubble elongation in the surfactant group versus 8.2% in controls (P < 0.05). Embolism shape change was coupled with surfactant-enhanced breakup into multiple smaller bubbles, which reabsorbed nearly 30% more rapidly than did parent bubbles in the control group (P < 0.05).