Transmural distribution of three-dimensional systolic strains in stunned myocardium.

BACKGROUND: Regional function in stunned myocardium is usually thought to be more depressed in the endocardium than the epicardium. This has been attributed to the greater loss of blood flow at the endocardium during ischemia. METHODS AND RESULTS: We measured transmural distributions of 3D systolic strains relative to local myofiber axes in open-chest anesthetized dogs before 15 minutes of left anterior descending coronary artery occlusion and during 2 hours of reperfusion. During ischemia, regional myocardial blood flow was reduced 84% at the endocardium and 32% at the epicardium (P<0.005, n=7), but changes in end-systolic fiber length from baseline were transmurally uniform. Relative to baseline, radial segments in stunned tissue were significantly thinner at the endocardium than the epicardium at end systole (24+/-5% versus 16+/-3%; P<0.05, n=8), consistent with previous reports. Unlike radial and cross-fiber segments, however, the increase of end-systolic fiber lengths in stunned myocardium had no significant transmural gradient (23+/-8% epicardium versus 21+/-4% endocardium). We also observed significant 3D diastolic dysfunction in the ischemic-reperfused region transmurally. CONCLUSIONS: Myocardial ischemia/reperfusion in the dog results in a significant transmural gradient of dysfunction between epicardial and endocardial layers in radial and cross-fiber segments, but not for fiber segments, despite a gradient in blood flow reduction during ischemia. Perhaps systolic fiber dysfunction rather than the degree of perfusion deficit during the preceding ischemic period may be the main determinant of myocardial dysfunction during reperfusion.

Flow-function relations during graded coronary occlusions in the dog: effects of transmural location and segment orientation.

OBJECTIVE: The sensitive relationship between regional myocardial perfusion and local systolic deformation during acute myocardial ischemia is not independent of the transmural location or segment orientation. The aim of this study was to determine the effects of fiber orientation and transmural location on the relationships between regional myocardial flow and three-dimensional systolic wall strain during graded coronary artery occlusions. METHODS: Transmural distributions of three-dimensional strain (by biplane radiography of implanted radiopaque markers) and myocardial blood flows (using fluorescent microspheres) were measured in the ischemic region during graded left anterior descending (LAD) coronary artery occlusions in 12 anesthetized dogs. RESULTS: Occlusion of the coronary artery did not significantly alter mean heart rate or end-systolic pressure. As flow decreased during graded occlusions, ischemia significantly changed systolic circumferential, longitudinal, radial, fiber and cross-fiber strains (p < 0.004). There was a significant effect of transmural position on circumferential, cross-fiber and radial strains, but not on fiber or longitudinal strains. Ischemia significantly altered all normal strains: circumferential, longitudinal, fiber, cross-fiber and radial. There was a strong interaction effect between transmural location and blood flow for circumferential, cross-fiber and radial strains, but not fiber or longitudinal strains. CONCLUSION: During non-transmural ischemia, there is evidence of strong transmural tethering in the cross-fiber direction, whereas the fiber-strain flow relation is independent of transmural position. Thus, whether the relationship between local myocardial bloodflow and systolic strain during acute ischemia is dependent on transmural location, depends on segment orientation.

Distributed mechanics of the canine right ventricle: effects of varying preload.

Fundamental questions in the mechanics of the right ventricle (RV) include: what are the distributions of diastolic and systolic strains across the RV epicardium and how do these strains change with increasing preload? Arrays (approximately 4 x 4 cm) of 25 to 30 lead markers were sutured to the epicardium of the RV anterior free wall in 6 open-chest, anesthetized dogs. Biplane cinéradiography (16 mm, 120 fps) was used to track marker positions throughout the cardiac cycle as loading conditions were altered by intravenous volume infusion. Continuous two-dimensional nonhomogeneous deformations were estimated across the region by fitting high-order finite element surfaces to the three-dimensional marker coordinates in successive ciné frames. End-systolic strains referred to end-diastole did not change with increasing preload, but did exhibit considerable longitudinal variation, e.g. the principal strain associated with maximal shortening (E1) was more than twice as great nearer the apex (E1 = -0.18 +/- 0.08) than in more basal (E1 = -0.09 +/- 0.05) regions. However, large amounts of lengthening occurred during diastolic inflation. End-diastolic extensional strains referred to an unloaded configuration were moderate at low pressure (E2 = 0.13 +/- 0.08) but increased to large values at high preloads (E2 = 0.28 +/- 0.11). End-diastolic strains also showed considerable longitudinal variation, i.e. near the base lengthening (E2 = 0.31 +/- 0.13) tended to be much greater than near the apex (E2 = 0.15 +/- 0.12). These results indicate that both diastolic sarcomere lengths and systolic sarcomere shortening increase in proportion to diastolic loading leaving end-systolic sarcomere strains unchanged.

Beta IG-H3, a novel secretory protein inducible by transforming growth factor-beta, is present in normal skin and promotes the adhesion and spreading of dermal fibroblasts in vitro.

We have previously identified a gene, beta ig-h3, which is highly induced in A549 cells (human lung adenocarcinoma) after growth arrest by transforming growth factor-beta. The beta ig-h3 gene encodes a 683-amino-acid secretory protein termed beta IG-H3, and treatment of several cell lines with transforming growth factor-beta results in increased secretion of beta IG-H3 into cell culture supernatants. In this report, we further characterize beta IG-H3 with respect to its synthesis and function. Primary human foreskin fibroblasts grown in monolayer culture produced beta IG-H3 mRNA and secreted beta IG-H3 protein into the growth media. Treatment of these cells with transforming growth factor-beta led to an increase in beta IG-H3 mRNA and protein. Cells grown on three-dimensional scaffolds secreted beta IG-H3 into the extracellular matrix, as judged by immunostaining with anti-beta IG-H3 antibodies. beta IG-H3 was also detected in normal human skin, especially in the papillary dermis. Finally, we show that recombinant beta IG-H3 supported attachment and spreading of dermal fibroblasts, suggesting that beta IG-H3 may function as an extracellular attachment protein in skin.

Cartilage production by rabbit articular chondrocytes on polyglycolic acid scaffolds in a closed bioreactor system.

Rabbit articular chondrocytes were seeded onto three-dimensional polyglycolic acid (PGA) scaffolds and placed into a closed bioreactor system. After 4 weeks of growth, meshes were examined for cartilage formation. Gross examination revealed solid, glistening material that had the appearance of cartilaginous tissue. Histologic examination revealed cell growth and deposition of extracellular matrix throughout the mesh with a less dense central core. Alcian blue and Safranin 0 staining showed deposition of glycosaminoglycans (GAGs). Immunostaining showed positive reactivity for type II collagen and chondroitin sulfate and no reactivity for type I collagen. Biochemical analysis showed collagen and GAG values to be 15% and 25% dry weight, respectively. Our results indicate that this type of system compares well with those previously described and should be useful for producing cartilage for evaluation in a clinical setting. (c) 1995 John Wiley & Sons, Inc.

Tgf-Beta promotes the growth of bovine chondrocytes in monolayer culture and the formation of cartilage tissue on three-dimensional scaffolds.

We have investigated the ability of transforming growth factor beta (TGF-beta) to promote the growth and differentiation of chondrocytes in monolayer and on three-dimensional scaffolds. Treatment of chondrocytes with TGF-beta and ascorbate individually stimulated the proliferation of bovine articular chondrocytes about 2-fold when cells were grown in monolayer culture: the combination of TGF-beta and ascorbate resulted in a 3-fold increase in cell number over a 72-h period. Peak stimulation with TGF-beta occurred at about 1.0 ng/ml: bFGF was slightly inhibitory in these assays. TGF-beta led to an increase in glycosaminoglycan synthesis as detected by Western blotting using anti-chondroitin sulfate antibodies. No significant change in collagen type II mRNA or protein was observed. When cells were grown on grown on three-dimensional scaffolds composed of polyglyocolic acid, TGF-beta treatment led to an increase in the size of the cartilage-like constructs produced. This was accompanied by increases in collagen and glycosaminoglycan deposition; immunohistochemical staining showed that the predominant collagen was type II. These results indicate that TGF-beta is capable of increasing the proliferation rate of chondrocytes in monolayer as well as increasing cartilage production on three-dimensional scaffolds and may find utility in the in vitro engineering of cartilage tissue.

Three-dimensional transmural mechanical interaction between the coronary vasculature and passive myocardium in the dog.

The "garden hose" effect of coronary perfusion on diastolic left ventricular (LV) mechanics has been proposed to cause changes in systolic function by altering diastolic sarcomere length. We measured transmural distributions of three-dimensional shape change using radiopaque markers implanted in the LV free wall of eight isolated arrested canine hearts as functions of coronary arterial perfusion pressure (Pp) and LV pressure (PLV) and related these deformations to the local muscle fiber architecture. Increased Pp from 0 to 110 mm Hg produced a 10% reduction in LV chamber volume (P < .01) and 25% to 40% decreases in local three-dimensional wall strain at matched PLV, indicating myocardial stiffening. Significant decreases in the magnitudes of local deformation occurred preferentially in the cross-fiber and radial directions (P < .02), with no change in fiber strain. This suggests that changing coronary Pp does not alter diastolic fiber length; hence, the Frank-Starling law may not mediate the Gregg effect. Since the myocardial microvessels are primarily oriented parallel to the muscle fibers, the observed myocardial stiffening occurs in the directions transverse to the microvessels rather than along their length. Local myocardial wall volume in the unloaded LV demonstrated a uniform 5% increase from the unperfused state to Pp of 50 mm Hg. With further increases in Pp up to 110 mm Hg, the change in regional wall volume from the unperfused state developed a substantial transmural gradient increasing by 7% at the epicardium and 15% at the subendocardium. This reflects a significant increase (P < .02) in intramyocardial coronary capacitance from epicardium to endocardium, which may be related to a transmural gradient in coronary distensibility or vascularity.

Vagally-mediated atrial fibrillation in dogs: conversion with bretylium tosylate.

We developed a stable, reproducible model of acute atrial fibrillation (sustained for up to 90 min) in open-chest, anesthetized dogs utilizing continuous bilateral stimulation of the cervical vagi and transient rapid atrial pacing. This model was then used to study the defibrillatory effects of bretylium tosylate. In five dogs, sequential runs of atrial fibrillation lasting 1-60 minutes were induced. A single, rapid, intravenous bolus of bretylium caused conversion of all episodes of atrial fibrillation (n = 31) after a mean of 33 seconds (range 15-151 sec). In 28 of 31 episodes (90%) conversion occurred within 40 seconds. The dose of bretylium was 2.5 mg/kg in all but one episode in which 5.0 mg/kg was given. We conclude that bilateral vagal stimulation produces a stable canine model of acute atrial fibrillation and that the arrhythmia is rapidly converted with intravenous bretylium.

Influence of the pericardium on left ventricular diastolic pressure-volume curves in dogs with sustained volume overload.

The pericardium is largely responsible for displacement of the left ventricular diastolic pressure-volume curve observed after acute volume loading in dogs. Likewise the pericardium has been considered likely to play a role in displacement of the curve in patients with acute cardiac failure and in shifts following manipulation of afterload. This study was designed to examine the influence of the pericardium on the diastolic pressure-volume relation of the left ventricle when volume load is more sustained, a setting relevant to observations made in patients with heart failure. We measured left ventricular pressure and volume in six conscious dogs with sustained volume overload (mean left ventricular end-diastolic pressure 21 mm Hg, left ventricular end-diastolic volume 149% of the upper limit of normal for our laboratory) produced by aortocaval shunt created 7 to 29 days earlier. Simultaneous left ventriculograms and pressures were obtained before and during nitroprusside infusion with the pericardium intact and in four dogs the studies were repeated 7 to 15 days after pericardiectomy. In all six dogs with intact pericardium, nitroprusside displaced the entire pressure-volume curve downward whereas after pericardiectomy, the pressure-volume data points obtained before and during nitroprusside infusion fell on a single curve. These results were similar to those previously reported for acute volume overload. Nitroprusside did not alter the time course of left ventricular pressure fall during the isovolumic period of diastole either before pericardiectomy (28.8 +/- 10.2 sec,-1, 28.4 +/- 11.9 sec-1) or after (28.8 +/- 6.7 sec-1, 26.1 +/- 7.2 sec-1). These data indicate that in dogs subjected to volume overload sustained for periods of up to 29 days, the pericardium affects the left ventricular diastolic pressure-volume curve and contributes to the elevation of left ventricular filling pressure through upward displacement of this curve.

Leukocyte capillary plugging in myocardial ischemia and reperfusion in the dog.

After several hours of ischemia an incomplete return of blood flow has been reported in brain, kidney, skeletal muscle, and heart. The mechanisms responsible for the no-reflow phenomenon have been unclear, and perivascular edema, platelet or red cell plugs, and interstitial hemorrhage have been implicated. In the present study evidence is provided that leukocyte entrapment in capillaries might contribute to no reflow. Leukocytes are large and stiff cells, which adhere to vascular endothelium naturally and are known to alter in their adherence properties under a variety of conditions. Accordingly, 11 open-chest dogs were studied, 1-5 hours after left anterior descending coronary artery occlusion. Reperfusion (9 dogs) at 75 mm Hg arterial pressure was accomplished with Ringer's lactate and carbon suspension as a marker for capillary patency. In non-ischemic tissue, 98% of the capillaries contained carbon, rare leukocytes, and few erythrocytes, whereas tissue from the distribution of the occluded artery was heterogeneous: 60% of the capillaries had no carbon, high hematocrits, and approximately one leukocyte per unbranched capillary; 40% demonstrated reflow and no leukocytes. A significant correlation between capillaries without carbon (no reflow) and the frequency of leukocytes remaining in these capillaries indicated that leukocytes were present in obstructed capillaries. Furthermore, the frequency of leukocytes remaining after the washout with lactate was ten times greater than in normal arrested heart muscle without washout. Our results suggest that progressive leukocyte capillary plugging during myocardial ischemia contributes to preventing full restoration of capillary flow upon reperfusion.

Opacification phase effects of angiographic contrast medium on left ventricular size and function: influence of initial ventricular volume and the pericardium.

Prior studies of the opacification-phase effects of left ventricular (LV) injection of angiographic contrast medium (CM) on LV size and function have yielded discrepant results. A sensitive ultrasonic technique was used to measure LV dimension (Di) in 10 open-chest dogs with physiologic heart rates, and injection of saline (S) was compared with injection of CM. The influence of initial LV volume (LVVi) and that of the pericardium were analyzed. Changes in contractile state (CS) were assessed from the end-systolic (ES) pressure-Di relation. With both low and high LVVi, S and CM produced indistinguishable early (beat 5) increases in end-diastolic (ED) Di and ESDi. EDDi and ESDi subsequently returned to baseline after S but remained elevated through beat 12 after CM. The latter increase was progressive only with low LVVi. Depression of CS was evident by beat 7 after CM, while pericardiectomy did not alter the response to CM. Thus, CM produces opacification-phase depression of CS and increases in LV size which are in part dependent in LVVi.

Cardiac norepinephrine stores and chronotropic reflex response after thoracotomy.

Although alterations in cardiac reflex activity, as well as in cardiac norepinephrine stores, have been shown following thoracic operations, the physiological relationship between these alterations has not been investigated. Cardiac norepinephrine stores were examined in 17 normal dogs, and in 16 dogs 1-6 wk after a right thoracotomy. The heart rate response to induced arterial hypotension (mean decrease 17-20 mmHg) prior to and following surgery was also examined in 10 dogs. A significant depression in right atrial norepinephrine stores from normal levels of 1.95 +/- 0.13 mug/g to 1.30 +/- 0.25 mug/g (33%) (P less than 0.05) was observed during the first wk following surgery. During the same period the heart rate response to a fall of 17-20 mmHg systolic arterial pressure was clearly reduced from 52.8 +/- 6.9 beats/min to 17.0 +/- 4.8 beats/min (67.8%; P less than 0.05). Right atrial norepinephrine stores at the time of the initial operation and when reflex responses were most markedly depressed demonstrated that the decrease in norepinephrine stores was paralleled by attenuation of the chronotropic response to hypotension.