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Biomedical subjects

D R Boughner

Publications and source records attributed to D R Boughner.

At least 37 records · Page 2Linked to original sources

Myocardial collagen changes and edema in rats with hyperdynamic sepsis.

OBJECTIVE: To determine if sepsis, which is accompanied by both systolic and diastolic myocardial dysfunction, involves changes in myocardial collagen, as myocardial collagen changes can affect both myocardial compliance and contractility. DESIGN: Prospective, randomized, controlled study. SETTING: Animal laboratory at a university-affiliated hospital. SUBJECTS: Male Sprague-Dawley rats, weighing 310 to 396 g. INTERVENTIONS: Cecal ligation and perforation (to induce sepsis) for 24 (n = 9) or 48 hrs (n = 9); sham laparotomy for 24 (n = 10) or 48 hrs (n = 9) with saline fluid resuscitation or normal control (n = 5) groups. MEASUREMENTS AND MAIN RESULTS: Collagen content and interstitial space were determined, using polarized light microscopy and a computer video densitometry system. At 24 and 48 hrs post surgery, heart rate and cardiac index were increased, and systemic vascular resistance index was decreased significantly in the sepsis vs. the sham rats. Collagen content was decreased significantly in the sepsis vs. the sham groups both at 24 and 48 hrs following surgery (1.83 +/- 0.79 [SD] % [24 hrs], 1.76 +/- 0.31% [48 hrs] vs. 2.83 +/- 0.73% [24 hrs], 2.25 +/- 0.72% [48 hrs]; p < .01). Interstitial space was increased significantly in the sepsis vs. the sham groups (13.9 +/- 3.5% [24 hrs], 15.6 +/- 5.2% [48 hrs] vs. 8.6 +/- 4.2% [24 hrs], 9.9 +/- 4.8% [48 hrs]; p < .01). CONCLUSIONS: Sepsis is accompanied by changes in myocardial collagen content and myocardial edema. These changes may contribute to the systolic and diastolic myocardial dysfunction, and particularly to the ventricular dilation, observed in sepsis.

Animals↗

Internal shear properties of fresh porcine aortic valve cusps: implications for normal valve function.

BACKGROUND AND AIMS OF THE STUDY: Several types of stress act on aortic heart valve tissue during the cardiac cycle. When closed the valve is subjected to primarily tensile stress due to the diastolic pressure, and upon opening bending stress occurs near the attachment with the aortic root and throughout the body of the cusps. Smooth bending requires internal tissue shearing. To measure the internal shear properties of the tissue a testing device was created which combined a high-precision linear actuator with a sensitive load cell. MATERIALS AND METHODS: Circular punch biopsy specimens from fresh porcine aortic valve cusps (n = 32) were examined. The shear stress versus shear strain characteristics were measured both in the circumferential (n = 17) and the radial (n = 13) direction, and the stress relaxation characteristics were also examined circumferentially (n = 15) and radially (n = 15). In addition seven specimens were tested repeatedly in both radial and circumferential directions for tissue isotropy. RESULTS: The results from the shear stress versus strain tests showed the tissue to behave non-linearly over the strain range between -0.9 and 0.9. The average moduli at the near zero strains were less than 300 Pa and increased to over 20 kPa at the extreme strains. The circumferential direction yielded slightly higher average moduli than the radial direction but this difference was not significant. The stress relaxation results indicated that valve tissue relaxation occurs in two distinct phases, an initial low slope region and a second high slope region with respective values of -7.5 log(s)-1 and -15 log(s)-1 and with no significant difference between test directions. CONCLUSIONS: Our results define and describe the pattern of internal shear properties of the aortic valve that are particularly important during the transition between the open and closed positions. This behavior pattern has particular application in the creation of accurate mathematical models of the valve tissue and may be important in understanding the mechanism of tissue failure in bioprosthetic valves.

Animals↗

Glutaraldehyde fixation alters the internal shear properties of porcine aortic heart valve tissue.

Glutaraldehyde fixation noticeably alters the mechanical properties of porcine aortic valve tissue, subsequently affecting the function and durability of these tissues when used as prosthetic heart valves. Traditional uniaxial tensile testing techniques do not fully define the mechanical properties and we have devised a new approach to examine the important shear properties of the tissue. Altered shear properties would change the response of the valve tissue as it flexes open and closed. An apparatus combining a high-precision linear actuator with a gram-sensitive load cell was used to measure the shear characteristics of circular punch specimens taken from the center of each valve cusp. The tissue parameters measured showed significant differences between the fixed and fresh tissues. Glutaraldehyde-fixed tissue (n = 16) was about 100 times as stiff as fresh tissue (n = 32) between shear strain values of 0 and 0.2. The fixed tissue also had stress relaxation rates about 60% those of the fresh cusps and had about 70% of the hysteresis loss seen in fresh tissue. These results demonstrated the significant effects of glutaraldehyde fixation on the properties of porcine aortic valve cusp when tested in shear. Such changes could lead to altered tissue function and may increase internal stresses during opening and closing, contributing to valve fatigue.

Animals↗

Bioprosthetic valve tissue viscoelasticity: implications on accelerated pulse duplicator testing.

Most of our knowledge of heart valve mechanics has been gained from low strain-rate studies much lower than physiologic levels. Using a high-speed materials testing system, we compared the low and high strain-rate viscoelastic behavior of porcine aortic valve cusps at extension rates of up to 40 mm/s. Circumferential and radial strips were stretched and then held in their stretched configuration to measure their "stress-relaxation" behavior. During low strain-rate stretching, only 6% of the initial stress dissipated or relaxed after 1 second, whereas 25% of the stress dissipated during high strain-rate stretching. This considerable difference in stress relaxation suggests a rate-dependent viscoelastic behavior that has not been accounted for in valve design and may have important implications for accelerated pulse testing. Even though the valve cusp is loaded for only 0.4 seconds during each heartbeat, at least 15% of the stress may relax over that period. During accelerated pulse testing, however, sufficient time may not be available to allow the tissue fibers to relax back to their natural state before the subsequent loading cycle, leading to a higher baseline preload. In addition, because valve tissue is not given sufficient time to relax before the next cycle, pulse testing subjects the valves to lower-magnitude cyclic stresses than does physiologic loading. Because both the baseline preload and the magnitude of cyclic stresses may lead to early fatigue failure, accelerated wear testing may either overestimate or underestimate valve durability. Clearly, the mechanism of stress-induced failure of biologic tissues must be elucidated before too much validity is placed on pulse duplicator studies.

Animals↗

A comparison of macroscopic lipid content within porcine pulmonary and aortic valves. Implications for bioprosthetic valves.

Lipid droplets have been demonstrated within both explanted porcine bioprostheses and normal porcine aortic valves. Because of the increasing interest in pulmonary valves as an allograft or xenograft aortic valve substitute, we examined the incidence and distribution of such lipid deposits in 50 porcine aortic valves and 50 matched porcine pulmonary valves. All 300 cusps were removed with surgical scissors and, under a dissecting microscope, the ventricularis layer was removed to expose the spongiosal layer. Macroscopic extracellular lipid droplets analyzed by means of a dissecting microscope with an eyepiece grid and stereology point-counting techniques to provide an area-density average spatial probability map for each cusp. Only 8% of porcine aortic valves were free of lipid, with the distribution of the lipids being 52% +/- 14% right coronary cusp, 90% +/- 8% left coronary cusp, and 68% +/- 13% noncoronary cusp. Of the pulmonary valves, 60% were free of lipid, with the incidence of lipids being 26% +/- 12% left cusp, 6% +/- 7% right cusp, and 12% +/- 9% anterior cusp. Subsequently, lipid cluster samples underwent thin-layer chromatography, which showed them to be phospholipids, oleic acid (fatty acid), triglycerides, and unesterified cholesterol. One primary mode of bioprosthetic valve failure is leaflet calcification. The similarity of distribution within the spongiosal layer between leaflet calcification and intrinsic cusp lipids suggests that the observed lipids might act as a nucleation site for calcification. The substantially lower incidence of lipid in pulmonary valves therefore may represent a potential benefit when these valves are considered for use as aortic valve replacements.

Animals↗

Oxygen uptake kinetics in cardiac transplant recipients.

Our purpose was to examine the gas exchange response to exercise in heart transplant (HT) patients and to characterize the O2 uptake kinetics (tau VO2) during successive square-wave on-transients from loadless cycling to moderate exercise. We hypothesized that with a slow heart rate response (and O2 transport limitation) O2 kinetics would be slowed but that with a repeated exercise initiated while the heart rate remained elevated the tau VO2 would be faster. Six male HT patients performed two ramp-function tests to determine peak O2 uptake (1.32 +/- 0.23 l/min) and ventilation threshold (1.02 +/- 0.16 l/min). Patients subsequently completed two repeats of a square-wave forcing function and repeated this on 2 days. Alveolar gas exchange was measured breath by breath. A monoexponential fit of signal-averaged data of the first exercise on-transient (between days) yielded a significantly slower tau VO2 in HT subjects than in healthy men (mean age 47 yr; n = 8) (77 +/- 26 vs. 45 +/- 4 s). With successive exercise (2nd transition) initiated while HR remained elevated the tau VO2 of HT patients was 46 +/- 17 s. The faster O2 kinetics of the second transition suggests that O2 delivery was enhanced and therefore that the tau VO2 may reflect bioenergetic processes controlling the rate of oxidative metabolism.

Blood Gas Analysis↗

Emergency echocardiographic evaluation of penetrating chest trauma.

Penetrating chest trauma is the most common cause of acute cardiac tamponade. Clinical recognition of this potentially life threatening condition may be difficult despite well described physical signs. Failure to repair the injury causing acute cardiac tamponade may result in sudden decompensation with poor clinical outcome. Emergent use of two-dimensional echocardiography can be an extremely valuable tool in evaluating the presence of cardiac tamponade and directing subsequent clinical management.

Adult↗

Enhanced detection of cardiac allograft arterial disease with intracoronary ultrasonographic imaging.

Intracoronary ultrasonographic imaging was performed in 60 patients 0.3 to 9 years (mean 2.9 +/- 1.9) after heart transplantation. By using a 1.8 mm intravascular ultrasonographic catheter, 192 (80%) of 240 angiographically visualized major epicardial coronary arteries (right, left main, anterior descending, and circumflex) were imaged by ultrasonography. Coronary luminal irregularities were detected in 15% of arteries by angiography compared with 34% by ultrasonography (p < 0.0001). The typical abnormality detected by ultrasonography consisted of crescentic and/or concentric intimal and medial thickening. Calcification in vascular lesions was rare (< 1% of arteries studied). Although the prevalence of angiographic abnormalities tended to be time dependent, ultrasonographic abnormalities were more strongly associated with donor age (normal, 22 +/- 8 years, vs abnormal, 33 +/- 10 years; p < 0.0001). Cardiac allograft coronary arterial disease is significantly underestimated by contrast angiography. Intravascular ultrasonography may provide a useful adjunct for identification and serial follow-up of this significant problem.

Adult↗

Stunned myocardium and myocardial collagen damage: differential effects of single and repeated occlusions.

It has been suggested that collagen loss and damage is responsible for the dysfunction seen in stunned myocardium. To test this hypothesis we compared collagen in canine hearts stunned by repeated occlusion with collagen in hearts stunned by a single occlusion. Regional contractile function was equally depressed in both groups: segment shortening at 1 hour after reperfusion averaged 37% +/- 9% versus 32% +/- 9% of preocclusion values in repeated and single occlusion models, respectively. Midmyocardial collagen content was not different in either single occlusion (10.5% +/- 0.4%) or repeated occlusion models (9.5% +/- 0.7%) when compared with nonischemic hearts (8.5 +/- 0.8%). Collagen damage, which was revealed with polarized light microscopy, was seen in 5 of 6 dogs after repeated occlusion but was not apparent after a single occlusion. Thus although both models of stunned myocardium produce similar dysfunction, there was no apparent collagen loss. Furthermore, collagen damage was only seen after repeated occlusion. Therefore it appears unlikely that collagen damage is a primary mechanism of stunned myocardium.

Animals↗

Cardiac function in aneurysmal subarachnoid haemorrhage: a study of electrocardiographic and echocardiographic abnormalities.

Electrocardiographic (ECG) changes are reported frequently after subarachnoid haemorrhage (SAH). The aim of this study was to investigate the functional significance of ECG changes by echocardiographic assessment of cardiac function. Forty-five patients with intracranial aneurysms were studied. All patients had a 12-lead ECG and a two-dimensional echocardiogram. After patients with an history of chronic cardiac disease (n = 4) were excluded, only four patients were found to have wall motion abnormalities. These patients had only minor ECG abnormalities, but severe neurological dysfunction. Conversely, patients with other ECG abnormalities including the deep inverted T waves associated usually with SAH, had normal echocardiograms. We conclude that the ECG is not an accurate predictor of myocardial function after SAH and that myocardial dysfunction is related more closely to severity of neurological condition.

Adolescent↗

Role of collagen in acute myocardial infarct expansion.

BACKGROUND: We sought to determine if damage to the myocardial collagen network was the cause of infarct expansion. METHODS AND RESULTS: Using polarized light microscopy, we examined sections from rat hearts obtained at 1, 2, 3, and 4 days after permanent coronary artery occlusion. Three features of the collagen network likely to be important in resisting infarct expansion were assessed: collagen quality, collagen quantity, and collagen organization. We observed a decrease in the number of normally birefringent collagen fibers in the infarct as early as 1 day after infarction. This decrease correlated significantly with time (r = -0.989, p less than 0.001). In addition, we found that the fewer normally birefringent collagen fibers present, the greater the degree of infarct expansion (assessed by measurement of total left ventricular cross-sectional area). At 4 days after infarction, we noted a loss of intermyocyte collagen struts and loss of interstitial space. These changes coincided with the onset of pronounced infarct expansion. The loss of collagen struts is consistent with the concept that expansion proceeds via slippage of myocytes previously tethered by the struts. The loss of interstitial space may represent the resolution of interstitial edema, which could further weaken the ventricular wall. CONCLUSIONS: The correlation of infarct expansion with collagen damage and the loss of support provided by collagen struts suggest that collagen is important in maintaining structural integrity after acute myocardial infarction.

Animals↗

Quantitative assessment of the age of fibrotic lesions using polarized light microscopy and digital image analysis.

Reliable histologic methods for gauging the maturity of fibrotic lesions are limited, making interventions in the healing process difficult to assess. As collagen ages there is enhanced birefringence due to increased molecular and fibrillar organization. The purpose of this study was to develop a microscopal technique to quantify this process and to determine its ability to distinguish scars of varying ages. Fibrosis in the rat gracilis muscle was studied 5 to 63 days after superficial injury. Sections were stained with picrosirius red and illuminated with monochromatic, polarized light. The microscope fields were digitized using a computer-video system yielding an image in which noncollagenous material was dark (gray level 0) and collagen was depicted by grey levels 1 to 255. In the fibrosis model used, the collagen area fraction plateaued at 80% by day 21. The median collagen grey level increased progressively as the scar aged. It is concluded that this histologic, nondestructive technique can reliably quantify age-related optical properties of fibrotic collagen and that this could be used to determine the maturity of fibrotic lesions.

Aging↗

Fibrosis in the transplanted heart and its relation to donor ischemic time. Assessment with polarized light microscopy and digital image analysis.

Long-distance procurement of cardiac allografts is commonly used to increase the supply of donor organs but has recently been associated with the development of impaired diastolic function. Therefore, the effect of the total ischemic duration on myocardial fibrosis was quantitatively evaluated in 36 cardiac transplant recipients in whom the ischemic time ranged from 70 to 363 (mean, 189 +/- 83) minutes. Interstitial collagen was quantified with polarization microscopy and digital image analysis in 115 endomyocardial biopsy specimens taken 5-10 days after surgery. The technique, developed for this study, showed excellent correlation with hydroxyproline analysis (r = 0.98, p less than 0.001). Collagen volume fraction in biopsy specimens from the transplanted hearts was significantly greater than that in biopsy samples from seven normal, age-matched autopsy hearts (4.7 +/- 1.9% vs. 2.9 +/- 0.6%, p less than 0.02). The degree of fibrosis correlated with the total ischemic time (r = 0.60, p less than 0.001). Donor age ranged from 10 to 51 years and did not correlate with the degree of fibrosis. No relation was found between the corresponding collagen content and right atrial pressure, pulmonary artery wedge pressure, or cardiac output measured at the time of biopsy. Myocyte damage was observed in eight of the 36 patients and was characterized by a striking loss of muscle birefringence. We conclude that cardiac allograft fibrosis may be identified shortly after transplantation and is dependent on the total ischemic duration.

Adult↗

The bovine pericardial xenograft: I. Effect of fixation in aldehydes without constraint on the tensile viscoelastic properties of bovine pericardium.

Tensile testing of tissue strips has been used to examine the effect of simple fixation in glutaraldehyde and formaldehyde on the viscoelastic properties of bovine pericardium. To assess tissue anisotropy, tissue strips were cut at 0 degree, 30 degrees, 60 degrees, and 90 degrees relative to the base-to-apex direction. Fresh anterior pericardium was modestly anisotropic, being least extensible in the base-to-apex direction; however, fixation removed this anisotropy. Fixation also produced a marked change in the response of the material to initial cyclic loading during preconditioning. Overall extensibility of the fixed material was significantly greater than that for the fresh tissue, consistent with a 10.7% shrinkage in aldehydes calculated from strain at fracture data. Reductions in stress relaxation and creep after fixation were noted as well, consistent with intrafibrillar crosslinking. Cyclic hysteresis and ultimate tensile strength were unaffected. Since the observed changes in the stress-strain response were largely attributable to shrinkage, control of shrinkage by physical means would allow for engineering modification of bovine pericardial mechanics for controlled anisotropy.

Aldehydes↗

An improved method for detecting and quantifying cardiac muscle disarray in hypertrophic cardiomyopathy.

The presence of muscle disarray in hypertrophic cardiomyopathy (HC), although well established, has only been semiquantitatively assessed. A quantitative method that uses polarized light microscopy is described. Hematoxylin and eosin-stained sections of ventricular septa from six HC patients and six normal hearts were examined. Cell orientations were measured in five regions from each section using a polarizing microscope with a rotating stage. Histograms of cell orientation angles were plotted and the mean and angular deviation of each sample were calculated. In normal hearts, cells were predominantly aligned parallel to each other. Orientation distributions were sharply peaked, with angular deviations ranging from 4 to 13 degrees. For HC, the sharp peak was lacking and angular deviations varied from 7 to 37 degrees; some distributions were bimodal. Areas in HC septa that appeared normal by gross inspection had abnormal orientation distributions. Polarized light microscopy provides an improved method of detecting and quantifying cellular disorganization in HC.

Cardiomyopathy, Hypertrophic↗

Measurements from light and polarised light microscopy of human coronary arteries fixed at distending pressure.

With the long term goal of improving our understanding of the mechanisms involved in coronary artery spasm, we have undertaken a two part study of the artery structure. We have made a comparison of the relative proportions of the different layers in proximal and distal regions of the main coronary arteries, and have quantitatively assessed their three dimensional structural fabric. Major coronary arteries from nine hearts were prepared for histological examination after fixation at a transmural pressure of 120 mm Hg. Measurements from 14 proximal and distal pairs of the cross sectioned arteries showed a dominant subendothelial layer, which diminished in thickness distally, with a small fraction of muscle cells interspersed with collagen. Three dimensional orientation measurements of the collagen and muscle components, which are birefringent, were obtained from one pair of segments from each of the left anterior descending, circumflex and right coronary arteries, using the polarising light microscope and Universal stage. Findings showed (1) a single circumferential order of adventitial collagen, with a mean circular standard deviation (CSD) of 22.3 degrees; (2) very highly ordered medial smooth muscle, mean CSD of 5.0 degrees (both findings are quantitatively similar between proximal and distal segments of artery, and between arteries); and (3) a multilayered fabric of collagen in the subendothelium in all vessel segments. The principal contributor to functional differences between proximal and distal regions may be the prominent and structurally varied subendothelial layer of the coronary arteries.

Adult↗

Analysis of healing after myocardial infarction using polarized light microscopy.

To better understand the healing process after permanent coronary artery occlusion in a canine model, the authors used polarized light microscopy. At 6 weeks after occlusion the scar collagen was mainly type I. Some regions of the scar contained a fiber lattice which appeared to be type III collagen. Collagen orientation was measured using a universal stage; subepicardial collagen was obliquely aligned (-14.0 +/- 3.5 degrees), midmyocardial collagen circumferentially aligned (1.4 +/- 0.4 degrees) and subendocardial collagen obliquely aligned (12.7 +/- 2.1 degrees). The molecular organization of scar collagen increased from 1 to 6 weeks after occlusion. Muscle cell disarray, similar to that in hypertrophic cardiomyopathy, was seen in the viable muscle adjacent to the scar. Such abnormal organization extended as far as 1 cm from the edge of the scar. The ability of polarized light microscopy to assess these different parameters from histologic sections demonstrates that it is a useful adjunct to other methods commonly used to study myocardial healing.

Animals↗