Closed-chest Animals Research Articles

A comprehensive and longitudinal cardiac magnetic resonance imaging study of the impact of coronary ischemia duration on myocardial damage in a highly translatable animal model.

We performed a comprehensive assessment of the effect of myocardial ischemia duration on cardiac structural and functional parameters by serial cardiac magnetic resonance (CMR) and characterized the evolving scar. CMR follow-up on the cardiac impact of time of ischemia in a closed-chest animal model of myocardial infarction with human resemblance is missing. Pigs underwent MI induction by occlusion of the left anterior descending (LAD) coronary artery for 30, 60, 90 or 120 min and then revascularized. Serial CMR was performed on day 3 and day 42 post-MI. CMR measurements were also run in a sham-operated group. Cellular and molecular changes were investigated. On day 3, cardiac damage and function were similar in sham and pigs subjected to 30 min of ischemia. Cardiac damage (oedema and necrosis) significantly increased from 60 min onwards. Microvascular obstruction was extensively seen in animals with ≥90 min of ischemia and correlated with cardiac damage. A drop in global systolic function and wall motion of the jeopardized segments was seen in pigs subjected to ≥60 min of ischemia. On day 42, scar size and cardiac dysfunction followed the same pattern in the animals subjected to ≥60 min of ischemia. Adverse left ventricular remodelling (worsening of both LV volumes) was only present in animals subjected to 120 min of ischemia. Cardiac fibrosis, myocyte hypertrophy and vessel rarefaction were similar in the infarcted myocardium of pigs subjected to ≥60 min of ischemia. No changes were observed in the remote myocardium. Sixty-minute LAD coronary occlusion already induces cardiac structural and functional alterations with longer ischemic time (120 min) causing adverse LV remodelling.

Apical closure device for full-percutaneous transapical structural and valve procedures with large-sized introducer sheaths: The final preclinical study.

Closed-chest transapical valve implantations (aortic, mitral, and tricuspid) and cardiac structural procedures requiring large-sized introducer sheaths cannot be safely performed with the available technology. We tested a self-expanding apical closure device in a closed-chest animal model, using large-sized introducer sheaths and human-sized animals to establish the technique for future tests in humans. Six human-sized pigs (mean weight: 89.7 ± 3.7 kg) received general anesthesia, intubation, and full heparinization (15,000 IU/animal; expected activated clotting time >200 s). Under fluoroscopy and multimodality imaging guidance with next-generation fusion imaging prototypes, a 15-cm long needle and a standard guidewire were percutaneously inserted under the xiphoidal aponeurosis and into the ventricular apex. After the exchange with a stiff guidewire, a 21-Fr introducer sheath for transapical procedures (outer diameter: 25-Fr) was placed in the left ventricle through the apex. The self-expanding closure device was inserted and deployed under fluoroscopic guidance while the 21-Fr sheath was gently removed. Hemodynamic conditions were monitored for 30 min and then the chest was opened to inspect the closure device and quantify the blood loss in the pericardium. Animals were killed and the hearts were removed and inspected. All six apical closure devices were successfully deployed without adverse events. No death, hemodynamic collapse, or cardiac tamponade occurred during the 30-minobservational period (mean systolic and diastolic pressures: 88 ± 11 and 58 ± 13 mmHg, respectively; mean heart rate: 60 ± 11 beats per minutes). Pre- and postdeployment (after protamine administration) mean activated clotting time was 541 ± 263 and 217 ± 62 s, respectively. The plugs provided good sealing with a mean of 27.2 ± 13.86 ml of blood lost in the pericardium. Postmortem inspection showed good plug fixation without myocardial damage. This self-expanding apical closure device successfully sealed the percutaneous access sites made with large-sized introducer sheaths in human-sized animals. This preclinical study suggests that transapical valve and structural procedures requiring large-sized introducer sheaths can be performed percutaneously.

Percutaneous Pericardial Resection: A Novel Potential Treatment for Heart Failure With Preserved Ejection Fraction.

People with heart failure and preserved ejection fraction develop increases in left ventricular (LV) end-diastolic pressures during exercise that contribute to dyspnea. In normal open-chest animal preparations, the pericardium restrains LV filling when central blood volume increases. We hypothesized that resection of the pericardium using a minimally invasive epicardial approach would mitigate the increase in LV end-diastolic pressure that develops during volume loading in normal and diseased hearts with the chest intact. Invasive hemodynamic assessment was performed at baseline and after saline load before and after pericardial resection in normal canines with open (n=3) and closed chest (n=5) and in a pig model with features of human heart failure and preserved ejection fraction with sternum intact (n=4). In closed-chest animals, pericardiotomy was performed using a novel subxiphoid procedure. In both experimental preparations of normal dogs, pericardiotomy blunted the increase in LV end-diastolic pressure with saline infusion, while enhancing the saline-mediated increase in LV end-diastolic volume. With chest intact in the pig model, percutaneous pericardial resection again blunted the increase in LV end-diastolic pressure secondary to volume expansion (+4±3 versus +13±5 mm Hg; P=0.014), while enhancing the saline-mediated increase in LV end-diastolic volume (+17±1 versus +10±2 mL; P=0.016). This proof of concept study demonstrates that pericardial resection through a minimally invasive percutaneous approach mitigates the elevation in LV filling pressures with volume loading in both normal animals and a pig model with diastolic dysfunction. Further study is warranted to determine whether this method is safe and produces similar acute and chronic hemodynamic benefits in people with heart failure and preserved ejection fraction.

A pig model of acute right ventricular afterload increase by hypoxic pulmonary vasoconstriction

BackgroundThe aim of this study was to construct a non-invasive model for acute right ventricular afterload increase by hypoxic pulmonary vasoconstriction. Intact animal models are vital to improving our understanding of the pathophysiology of acute right ventricular failure. Acute right ventricular failure is caused by increased afterload of the right ventricle by chronic or acute pulmonary hypertension combined with regionally or globally reduced right ventricular contractile capacity. Previous models are hampered by their invasiveness; this is unfortunate as the pulmonary circulation is a low-pressure system that needs to be studied in closed chest animals. Hypoxic pulmonary vasoconstriction is a mechanism that causes vasoconstriction in alveolar vessels in response to alveolar hypoxia. In this study we explored the use of hypoxic pulmonary vasoconstriction as a means to increase the pressure load on the right ventricle.ResultsPulmonary hypertension was induced by lowering the FiO2 to levels below the physiological range in eight anesthetized and mechanically ventilated pigs. The pigs were monitored with blood pressure measurements and blood gases. The mean pulmonary artery pressures (mPAP) of the animals increased from 18.3 (4.2) to 28.4 (4.6) mmHg and the pulmonary vascular resistance (PVR) from 254 (76) dyns/cm5 to 504 (191) dyns/cm5, with a lowering of FiO2 from 0.30 to 0.15 (0.024). The animals’ individual baseline mPAPs varied substantially as did their response to hypoxia. The reduced FiO2 level yielded an overall lowering in oxygen offer, but the global oxygen consumption was unaltered.ConclusionsWe showed in this study that the mPAP and the PVR could be raised by approximately 100% in the study animals by lowering the FiO2 from 0.30 to 0.15 (0.024). We therefore present a novel method for minimally invasive (closed chest) right ventricular afterload manipulations intended for future studies of acute right ventricular failure. The method should in theory be reversible, although this was not studied in this work.

Chronic Oral Amiodarone but not Dronedarone Therapy Increases Ventricular Defibrillation Threshold During Acute Myocardial Ischemia in a Closed-Chest Animal Model

Dronedarone, a recently approved antiarrhythmic drug, has been shown to have fewer side effects than amiodarone, particularly with regard to thyroid and neurologic events. Since the effects of either drug on ventricular defibrillation threshold during ischemia are unknown, the aim of this study was to compare the effects of dronedarone and amiodarone on defibrillation efficacy during ischemia in a closed-chest animal model. Dronedarone (30 mg·kg·d) and amiodarone (20 mg·kg·d) were administered orally for 3 weeks to 19 and 21 pigs, respectively. A control group (no treatment) comprised 19 pigs. A 2-lead endovascular defibrillation system was used. Each biphasic shock was delivered after 8 seconds of ventricular fibrillation. A step-up/step-down protocol was used to calculate mean defibrillation threshold before and 10 minutes after coronary artery occlusion using an angioplasty balloon in the left descending artery. At basal state, defibrillation threshold did not differ between the control (20.8 ± 4.8 J), amiodarone (21.2 ± 2 J), and dronedarone (19.5 ± 3 J) groups. After ischemia, the amiodarone group had a significantly higher defibrillation threshold than the control group (29.6 ± 3 J vs. 21.8 ± 5 J, respectively; P = 0.015), but the dronedarone (22.8 ± 4 J) and control groups had similar defibrillation threshold values. These data indicate that oral dronedarone treatment, unlike oral amiodarone, does not affect defibrillation threshold during ischemia in pigs.

Closed-chest animal model of chronic coronary artery stenosis. Assessment with magnetic resonance imaging

To evaluate the consequences of chronic non-occlusive coronary artery (CA) stenosis on myocardial function, perfusion and viability, we developed a closed-chest, closed-pericardium pig model, using magnetic resonance imaging (MRI) as quantitative imaging tool. Pigs underwent a percutaneous copper-coated stent implantation in the left circumflex CA (n = 19) or sham operation (n = 5). To evaluate the occurrence of myocardial infarction, cardiac troponin I (cTnI) levels were repetitively measured. At week 6, CA stenosis severity was quantified with angiography and cine, first-pass and contrast-enhanced MRI were performed to evaluate cardiac function, perfusion and viability. In the stenting group, cTnI values significantly increased at day 3 and day 5 (P = 0.01), and normalized at day 12. At angiography, 13/19 stented pigs had a stenosis >75%. Mean degree of CA stenosis was 91 +/- 4%, range 83-98%. At contrast-enhanced MRI, mean infarct size was 7 +/- 6%, range 0.7-18.4%. Five of the 6 pigs with stenosis <75% had no infarction. Stented pigs showed significantly higher Left-ventricular volumes and normalized mass (P < 0.05), and lower ejection fraction (P = 0.03) than the sham pigs. Both wall thickening and myocardial perfusion were significantly lower in animals with at least one segment >50% infarct (23 +/- 8%; 0.05 +/- 0.01 a.u./s) and animals with only <50% infarct segments (29% +/- 12%; 0.07 +/- 0.02 a.u./s), than sham pigs (52 +/- 6%; 0.10 +/- 0.03 a.u./s) (P < 0.001; P < 0.05). This minimally-invasive animal model of chronic, non-occlusive CA stenosis, presenting a mixture of perfusion and functional impairment and a variable degree of myocardial necrosis, can be used as substitute to study chronic myocardial hypoperfusion.

The Impact of Acute Myocardial Ischemia on the Ventricular Defibrillation Threshold During Chronic Oral Azimilide Therapy

The effects of chronic oral azimilide therapy on the ventricular defibrillation threshold (DFT) during ischemia are unknown. The effects of azimilide on defibrillation efficacy under ischemic condition were investigated in a closed-chest animal model. Azimilide (20 mg/kg/d) was administered orally for 7 days to 10 pigs (20 to 25 kg). The control group (no treatment) comprised 15 pigs. A 2-lead defibrillation system was used. Each shock was delivered after 8 seconds of ventricular fibrillation. A step-up and step-down protocol was used to calculate mean DFT before and 10 minutes after coronary artery occlusion using an angioplasty balloon in the left descending artery. The basal DFT of the azimilide group did not differ from controls (20.8 +/- 4.8 versus 18.8 +/- 2.8; P = 0.33). After ischemia, the mean DFT of the azimilide-treated animals was similar to controls (21.8 +/- 5.2 versus 23.2 +/- 3.8 J; P = 0.54), despite significant lengthening of ventricular repolarisation (428.2 +/- 51.8 versus 494.1 +/- 46.6 msec; P = 0.005) and significant prolongation of the ventricular fibrillation cycle length (85.1 +/- 13 versus 104.7 +/- 24 msec; P < 0.04). Chronic oral azimilide treatment does not affect the DFT at baseline or during acute myocardial ischemia.

Development of an Electrophysiology (EP)-Enabled Intracardiac Ultrasound Catheter Integrated With NavX 3-Dimensional Electrofield Mapping for Guiding Cardiac EP Interventions

We have developed an integrated high-resolution intracardiac echocardiography (ICE) catheter for electrophysiology (EP) testing, which can be coregistered in 3-dimensional space with EP testing and ablation catheters using electrofield sensing. Twelve open-chest pigs (34-55 kg) and 3 closed-chest pigs were studied. After introduction from the jugular or femoral venous locations, the 9F side-looking, highly steerable (0 degrees -180 degrees), 64-element array catheters could be manipulated easily throughout the right side of the heart. Multisite cardiac pacing was performed for assessing left ventricular (LV) synchrony using tissue Doppler methods. Also, in the open-chest pigs, right atrial (RA) and right ventricular (RV) ablations were performed with a separate radio frequency catheter under fluoroscopic guidance and visualized with ICE to characterize the changes. In the 3 closed-chest pigs, electrofield NavX 3-dimensional coregistration (St Jude Medical Corp, Minneapolis, MN) allowed us to test whether this additional feature could shorten the time necessary to perform 4 targeted ablations in each animal while imaging the ablation catheter and the adjacent region by ICE. Intracardiac anatomy, tricuspid, aortic, pulmonary, and mitral valve function, and pulmonary vein flow were all imaged reproducibly from scanning locations in the RA or RV in all animals, along with assessment of cardiac motion and the effects of multisite pacing. Three-dimensional electrofield displays detailed the spatial relationship between the ICE catheter and ablation catheters such that the time to visualize and ablate 4 sites in each of the 3 closed-chest animals was reduced. This new technology is a first step in the integration of ICE with EP procedures.

Alterations in Lateral Left Ventricular Wall Transmural Strains During Acute Circumflex and Anterior Descending Coronary Occlusion

Increased circumferential-radial shear in the midlateral left ventricle adjacent to ischemic myocardium has been observed during acute midcircumflex ischemia in open-chest animals. Extending this work, we studied transmural strains in closed-chest animals during acute proximal-circumflex (pCX) and proximal-left anterior descending (pLAD) occlusions. Six sheep had radiopaque markers implanted to silhouette the left ventricle and measure regional systolic fractional area shortening; three transmural bead columns were inserted into the midlateral wall for transmural myocardial strain analysis. After 8 weeks, three-dimensional marker coordinates were obtained using biplane videofluoroscopy, both before and during separate 1-minute pLAD and pCX balloon occlusions. Systolic strains were assessed along circumferential, longitudinal, and radial axes, and then transformed into fiber strains using quantitative microstructural measurements. Acute pLAD occlusion and pCX occlusion caused similar hemodynamic insults. Systolic fractional area shortening revealed that the beads were in the ischemic territory during pCX occlusion, but adjacent to the ischemic myocardium during pLAD occlusion. Transmural circumferential strain and fiber shortening fell in the ischemic region during pCX occlusion, but remained normal when adjacent to the ischemic myocardium during pLAD occlusion. Circumferential-radial shear strain increased in the lateral left ventricle during pCX occlusion, but reversed in this same region during pLAD occlusion. Longitudinal-radial shear also decreased during pLAD occlusion. Reversal of lateral wall circumferential-radial shear and decreased longitudinal-radial shear during acute pLAD occlusion reflects altered mechanical interaction between ischemic and nonischemic myocardium. Increased circumferential-radial shear during pCX occlusion also reflects mechanical interaction. The direction of circumferential-radial shear deformation depends on the location of the adjacent ischemic territory.

Contribution of mitral annular dynamics to LV diastolic filling with alteration in preload and inotropic state

Mitral annular (MA) excursion during diastole encompasses a volume that is part of total left ventricular (LV) filling volume (LVFV). Altered excursion or area variation of the MA due to changes in preload or inotropic state could affect LV filling. We hypothesized that changes in LV preload and inotropic state would not alter the contribution of MA dynamics to LVFV. Six sheep underwent marker implantation in the LV wall and around the MA. After 7-10 days, biplane fluoroscopy was used to obtain three-dimensional marker dynamics from sedated, closed-chest animals during control conditions, inotropic augmentation with calcium (Ca), preload reduction with nitroprusside (N), and vena caval occlusion (VCO). The contribution of MA dynamics to total LVFV was assessed using volume estimates based on multiple tetrahedra defined by the three-dimensional marker positions. Neither the absolute nor the relative contribution of MA dynamics to LVFV changed with Ca or N, although MA area decreased (Ca, P < 0.01; and N, P < 0.05) and excursion increased (Ca, P < 0.01). During VCO, the absolute contribution of MA dynamics to LVFV decreased (P < 0.001), based on a reduction in both area (P < 0.001) and excursion (P < 0.01), but the relative contribution to LVFV increased from 18 +/- 4 to 45 +/- 13% (P < 0.001). Thus MA dynamics contribute substantially to LV diastolic filling. Although MA excursion and mean area change with moderate preload reduction and inotropic augmentation, the contribution of MA dynamics to total LVFV is constant with sizeable magnitude. With marked preload reduction (VCO), the contribution of MA dynamics to LVFV becomes even more important.

Imaging of the pulmonary circulation in the closed-chest rat using synchrotron radiation microangiography

Structural changes of the pulmonary circulation during the pathogenesis of pulmonary arterial hypertension remain to be fully elucidated. Although angiography has been used for visualizing the pulmonary circulation, conventional angiography systems have considerable limitations for visualizing small microvessels (diameters < 200 microm), particularly within a closed-chest animal model. In this study we assess the effectiveness of monochromatic synchrotron radiation (SR) for microangiography of the pulmonary circulation in the intact-chest rat. Male adult Sprague-Dawley rats were anesthetized, and a catheter was positioned within the right ventricle, for administering iodinated contrast agent (Iomeron 350). Subsequently, microangiography of pulmonary arterial branches within the left lung was performed using monochromatic SR. Additionally, we assessed dynamic changes in vessel diameter during acute hypoxic (10% and 8% O2 for 4 min each) pulmonary vasoconstriction (HPV). Using SR we were able to visualize pulmonary microvessels with a diameter of <100 microm (the 4th generation of branching from the left axial artery). Acute hypoxia caused a significant decrease in the diameter of all vessels less than 500 microm. The greatest degree of pulmonary vasoconstriction was observed in vessels with a diameter between 200 and 300 microm. These results demonstrate the effectiveness of SR for visualizing pulmonary vessels in a closed-chest rat model and for assessing dynamic changes associated with HPV. More importantly, these observations implicate SR as an effective tool in future research for assessing gross structural changes associated with the pathogenesis of pulmonary arterial hypertension.

Direct visualization of coronary sinus ostium and branches with a flexible steerable fiberoptic infrared endoscope

Placement of electrophysiology catheters and pacing leads in the coronary sinus is challenging in some patients, particularly those with dilated cardiomyopathy. We hypothesized that cannulation of the coronary sinus and its branches can be facilitated by direct visualization. This study reports our experience with navigation into and within the coronary sinus in a closed-chest animal preparation, using a flexible steerable fiberoptic infrared endoscope that allows visualization through flowing blood. The purpose of this study was to assess the feasibility of direct visualization of endocardial structures through infrared endoscopy. Internal jugular venous access was obtained in 10 healthy mongrel dogs (weight 35-45 kg). The infrared endoscope (2900 fiber imaging bundle, wavelength 1,620 nm, frame rate 10-30/s, 320 x 256 pixels) was advanced to the coronary sinus ostium and branches by direct visualization of anatomic landmarks, such as the tricuspid valve and inferior vena cava. Localization was confirmed by fluoroscopy, contrast injection, and pathologic examination. Structures such as the tricuspid valve and inferior vena cava were visualized at distances of 1 to 2 cm, allowing successful coronary sinus identification and engagement in all 10 dogs. Coronary sinus branch images closely resembled pathologic findings. Direct visualization of the coronary sinus ostium and branches is possible through infrared endoscopy. This technique likely will facilitate coronary sinus engagement and navigation for pacing lead and catheter placement.

Effects of mechanical ventilation at low lung volume on respiratory mechanics and nitric oxide exhalation in normal rabbits

Lung mechanics, exhaled NO (NOe), and TNF-alpha in serum and bronchoalveolar lavage fluid were assessed in eight closed and eight open chest, normal anesthetized rabbits undergoing prolonged (3-4 h) mechanical ventilation (MV) at low volume with physiological tidal volumes (10 ml/kg). Relative to initial MV on positive end-expiratory pressure (PEEP), MV at low volume increased lung quasi-static elastance (+267 and +281%), airway (+471 and +382%) and viscolelastic resistance (+480 and +294%), and decreased NOe (-42 and -25%) in closed and open chest rabbits, respectively. After restoration of PEEP, viscoelastic resistance returned to control, whereas airway resistance remained elevated (+120 and +31%) and NOe low (-25 and -20%) in both groups of rabbits. Elastance remained elevated (+23%) only in closed-chest animals, being associated with interstitial pulmonary edema, as reflected by increased lung wet-to-dry weight ratio with normal albumin concentration in bronchoalveolar lavage fluid. In contrast, in 16 additional closed- and open-chest rabbits, there were no changes of lung mechanics or NOe after prolonged MV on PEEP only. At the end of prolonged MV, TNF-alpha was practically undetectable in serum, whereas its concentration in bronchoalveolar lavage fluid was low and similar in animals subjected or not subjected to ventilation at low volume (62 vs. 43 pg/ml). These results indicate that mechanical injury of peripheral airways due to their cyclic opening and closing during ventilation at low volume results in changes in lung mechanics and reduction in NOe and that these alterations are not mediated by a proinflammatory process, since this is expressed by TNF-alpha levels.

Lung biopsy with a 12-gauge cutting needle is possible using an insertion sheath in animal models 1

The volume of lung tumor core biopsy specimens has been restricted because of concerns for complications such as bleeding and air leakage. In this animal experiment, we investigated the possibility of larger bore biopsies through the peripheral lung parenchyma. Lung biopsy was done in male domestic pigs (n= 4) under thoracotomy. A single biopsy using a 12-gauge cutting biopsy needle was done with sheath (sheath group, eight biopsies) or without sheath (nonsheath group, eight biopsies). After biopsy, bleeding time, bleeding amount, and positive airway pressure causing air leakage from the insertion site was compared between groups (Mann-Whitney U test). To observe long-term effects in closed-chest animals, percutaneous lung biopsy with the use of a sheath was carried out percutaneously in male beagles (n = 9). The animals were observed for 3 weeks. In the pigs (sheath group) after biopsy, bleeding flowed through the sheath and formed a sheath-molded fibrin plug that secured the insertion site. Bleeding time and amount decreased significantly in the sheath group compared with the nonsheath group (115 +/- 108 versus 295 +/- 150 seconds, P = .018, and 37 +/- 41 versus 98 +/- 72 grams, P= .027, respectively). Air leakage pressure was significantly higher in the sheath group compared with the nonsheath group (37 +/- 6 versus 18 +/- 5 cmH2O, P = .001). In the beagles, no complications such as pneumothorax, hemothorax, or airway bleeding was apparent. Although we have not evaluated lung tumor biopsy per se, lung tumor biopsy with a 12-gauge cutting needle may be possible with a use of a sheath.

Pulmonary arterial distension and vagal afferent nerve activity in anaesthetized dogs.

Distension of the main pulmonary artery and its bifurcation are known to result in a reflex vasoconstriction and increased respiratory drive; however, these responses are observed at abnormally high distending pressures. In this study we recorded afferent activity from pulmonary arterial baroreceptors to investigate their stimulus-response characteristics and to determine whether they are influenced by physiological changes in intrathoracic pressure. In chloralose-anaesthetized dogs, a cardiopulmonary bypass was established, the pulmonary trunk and its main branches were vascularly isolated and perfused with venous blood at pulsatile pressures designed to simulate the normal pulmonary arterial pressure waveform. Afferent slips of a cervical vagus were dissected and nerve fibres identified that displayed discharge patterns with characteristics expected from pulmonary arterial baroreceptors. Recordings were obtained with (a) chest open (b) chest closed and resealed, and (c) with phasic negative intrathoracic pressures in the resealed chest. Pressure-discharge characteristics obtained in the open-chest animals indicated that the threshold pulmonary pressure (corresponding to 5% of the overall response) was 17.1 +/- 2.9 and the inflexion point of the curve was 29.2 +/- 3.3 mmHg (mean +/-S.E.M). In closed-chest animals the threshold and inflexion pressures were reduced to 12.0 +/- 1.7 and 20.7 +/- 1.8 mmHg. Application of phasic negative intrathoracic pressures further reduced the threshold and inflexion pressures to 9.5 +/- 1.2 mmHg (P < 0.05 vs. open) and 14.7 +/- 0.8 mmHg (P < 0.003 vs. open and P < 0.02 vs. atmospheric). These results indicate that under physiological conditions, with closed-chest and phasic negative intrathoracic pressure changes similar to those associated with normal breathing, activity from pulmonary baroreceptors is obtained at physiological pulmonary arterial pressures in intact animals.

Pharmacologic pre-conditioning and controlled reperfusion prevent ischemia–reperfusion injury after 30 minutes of hypoxia/ischemia in porcine hearts

Background Hearts from non–heart-beating organ donors are not transplanted because of risk of ischemia-reperfusion injury. We tested whether pharmacologic pre-conditioning with adenosine and the Na +/H + exchanger inhibitor, cariporide, combined with controlled reperfusion, would prevent injury in porcine hearts that had sustained 30 minutes of hypoxia/ischemia in closed-chest animals. Methods Hearts from Yorkshire pigs (100 kg) were studied in 3 groups. Group 1 (control) hearts were surgically removed while beating. Group 2 hearts were harvested from animals made hypoxic by discontinuing mechanical ventilation for 30 minutes. Group 3 hearts were hypoxic as in Group 2, but these animals received adenosine (40 mg) and cariporide (400 mg) 10 minutes before stopping ventilation. Cardiac function in all groups was assessed ex vivo in a working heart apparatus in which pressure and flow measurements were made over 3 hours. Controlled reperfusion in Group 3 hearts used leukocyte-depleted blood perfusate containing free radical scavengers. Myocardial injury was assessed on the basis of perfusate creatine phosphokinase activity and histopathologically determined injury score. Results Groups 1 and 3 hearts could be resuscitated to perform work equivalently during the entire reperfusion period and showed positive responses to increases in pre-load and norepinephrine. Group 2 hearts could not perform work. After 3 hours, Group 2 hearts showed significantly higher creatine phosphokinase and histopathologic injury scores compared to with Groups 1 and 3, which were not significantly different from each other. Conclusions Pharmacologic pre-conditioning and controlled reperfusion effectively protect non-beating porcine hearts from injury after 30 minutes of hypoxia/ischemia in situ.

New efficient catheter-based system for myocardial gene delivery.

Manipulating gene expression in the failing heart has therapeutic promise, but until now efficient and homogeneous cardiac gene delivery has required an open-chest approach. This study examines the hypothesis that vector delivery promoted by echo contrast microbubbles will be maximized by injection of the vectors into the aortic root with brief balloon occlusion above the sinuses, while at the same time prolonging diastole and vasodilating with acetylcholine (ACh) to maximize coronary exposure. After incubation with albumin-coated perfluorocarbon microbubbles, an adenovirus encoding a reporter gene was infused into the aortic root of rats. To maximize delivery, the aortic root was transiently occluded with a balloon catheter during a brief ACh-induced asystole. Ultrasound was used to image the delivery and disrupt the microbubbles. Aortic occlusion with concomitant ACh increased myocardial gene expression for virus + microbubbles by >2.5-fold, from 925+/-165 to 2358+/-376 relative units (RU; P<0.01). This delivery system also produced substantial expression with vector alone (1473+/-549 RU). All uptakes were significant compared with 433+/-332 RU without virus. An adenoviral delivery system combining echo contrast with a catheter-based technique to maximize coronary perfusion increases gene delivery compared with echo contrast alone. This novel method permits efficient percutaneous gene delivery in closed-chest animals.

Xenon produces minimal haemodynamic effects in rabbits with chronically compromised left ventricular function

Xenon has only minimal haemodynamic side-effects on normal myocardium and might be a preferable anaesthetic agent for patients with heart failure. We studied the haemodynamic changes caused by 70% xenon in rabbits with chronically compromised left ventricular (LV) function. Anaesthetized rabbits were thoracotomized and a major coronary artery was ligated to induce ischaemic heart disease. Nine weeks later, rabbits were again anaesthetized (ketamine/propofol), and haemodynamics were measured during inhalation of 70% xenon using echocardiography [LV end-diastolic dimension (LVedD), fractional shortening (FS), velocity of circumferential fibre shortening (VcF), ejection fraction (EF)] in closed-chest animals. Subsequently, rabbits were thoracotomized and instrumented for measurement of LV pressure (tip manometer), LV dP/dtmax and cardiac output (ultrasonic flow probe). Haemodynamics were recorded again during inhalation of 70% xenon. All rabbits had compromised LV function 9 weeks after coronary artery ligation. Mean LVedD increased from 12.9 (SD 0.9) mm to 17.1 (0.4) mm; EF decreased from 73 (9) to 64 (8)%; FS decreased from 36 (7) to 29 (5)%; VcF decreased from 28.9 (6.8) to 17.6 (4.7) mm s(-1); all P<0.05. Inhalation of 70% xenon had no effect on haemodynamics in closed-chest rabbits, as measured by echocardiography. After invasive instrumentation, small decreases in LV pressure from 78 (20) to 72 (19) mm Hg, LV dP/dtmax from 3081 (592) to 2633 (503) mm Hg s(-1) and cardiac output from 239 (69) to 225 (71) ml min(-1) were observed during xenon inhalation (all P<0.05). These data show that xenon has only minimal negative inotropic effects in rabbits with LV dysfunction after coronary artery ligation.

Quantification of the spectrum of changes in regional myocardial function during acute ischemia in closed chest pigs: An ultrasonic strain rate and strain study

The objectives of this study were to define the spectrum of regional myocardial function changes during acute ischemia in closed chest animals by using newly developed ultrasonic strain rate and strain indexes derived from regional color Doppler myocardial imaging (CDMI) velocity data. Myocardial ischemia was induced in 18 pigs either with acute total 20-second occlusions (group 1, n = 12) or graded hypoperfusion (40 to 0 mL/min, group 2, n = 6) of the circumflex coronary artery. In addition, a dobutamine challenge (5 to 10 μg/kg per minute) was performed during sustained subtotal ischemia (10 mL/min) in group 2. CDMI acquisitions with parasternal views monitored the myocardial posterior wall function. Regional radial strain rate and strain (ϵr) were measured for systole, isovolumic relaxation, early diastole, and atrial filling, respectively. During total and graded ischemia, ϵr profiles were consistently modified, showing a delayed onset and a decrease in regional systolic thickening as well as increased postsystolic thickening. Radial strain rate and ϵr indexes decreased consistently during systole and early diastole and increased during isovolumic relaxation. End-systolic ϵr could differentiate total ischemia from severe hypoperfusion (10 mL/min), decreasing from 32% ± 8% to 16% ± 5% (versus 60% ± 10% at baseline). During dobutamine infusion (10 μg/kg per minute), end-systolic ϵr tended to decrease from 27% ± 5% to 18% ± 11%, whereas postsystolic thickening increased by 2-fold (P <.05). The combined analysis of regional deformation characteristics and global cardiac event timing derived from CDMI data can identify and quantify regional function changes induced by experimental acute ischemia in closed chest pigs. This would appear to be a potentially promising new noninvasive approach to the clinical evaluation of ischemia-induced changes in segmental myocardial function. (J Am Soc Echocardiogr 2001;14:874-84.)

Changes in systolic and postsystolic wall thickening during acute coronary occlusion and reperfusion in closed-chest pigs: Implications for the assessment of regional myocardial function

The aim of the study was to characterize the impact of short-lived total coronary occlusions in closed-chest pigs on radial wall thickening within the “at-risk” myocardial segment by using gray-scale M-mode echocardiography. Twelve pigs underwent a series of 20-second total circumflex coronary artery occlusions with an angioplasty balloon. Myocardial thickening/thinning indexes were continuously monitored before ischemia, during ischemia, and on reperfusion by high-resolution M-mode recordings of the posterior wall obtained from parasternal views. The timing of regional events was compared with global systolic time intervals derived from the color Doppler myocardial imaging velocity data. Each occlusion induced a rapid decrease in end-systolic thickening (ϵ ES), closely paralleled by an increase in postsystolic thickening in the ischemic segment. After 20 seconds of ischemia, ϵ ES decreased by −86% and postsystolic thickening increased by +100%, whereas maximal thickening decreased only by −34% in comparison with preocclusion values. All wall thickening parameters returned to baseline after 15 seconds of reperfusion. During acute total ischemia in a closed-chest animal model, the changes in regional myocardial function were best characterized by the combined analysis of systolic and postsystolic thickening abnormalities and by their respective timings relative to global cardiac events markers. (J Am Soc Echocardiogr 2001;14:691-7.)