Low frequency dynamic viscoelastic properties of human mitral valve tissue.

The dynamic viscoelasticity of the anterior leaflet of human mitral valves was investigated by subjecting the ventricular aspect of membranous samples of the tissue to sinusoidal fluid pressures. The frequency range of the stressing function used was from 0.5 Hz to 5 Hz. The storage modulus of the tissue was found to be independent of the stressing frequency and an average value of 2.55 (SE = 0-12) X 10(8) dyn cm-2 was observed. Losses were small and the phase shift between stressing function and resulting strain and hence the loss modulus increased with frequency. For the frequency range investigated the phase shift varied from 0.04 to 0.06 rad and the loss modulus was of the order of 10(7) dyn cm-2. From the observed data it was concluded that any tissue substitute used in mitral valve replacement should be rather inextensible and have a low loss modulus. Also under normal physiological conditions, the mitral valve cannot bulge into the left atrium during peak ventricular systole and hence events in the cardiac cycle, such as the presence of the atrial pressure 'c' wave, that involve the distensibility of the valve need to be re-examined.

Contrasting structure of the saphenous vein and internal mammary artery used as coronary bypass vessels.

OBJECTIVES: To report quantitatively on the three-dimensional layered organization of the collagen and smooth muscle component of the two most successful vessels for coronary bypass-the internal mammary artery (IMA) and the long saphenous vein (SV). Our aim was to provide an explanation for the differential structural stiffness of these two vessels (both functioning at arterial pressures in their new environment), and how they might be susceptible to endothelial thickening. METHODS: Eleven human saphenous veins and 23 internal mammary arteries were fixed at arterial distending pressure of 110 mmHg, and were sectioned in cross-section at 7 microns thickness. A subset of these was also sectioned tangentially. Measurements of the three-dimensional alignment of collagen and smooth muscle fibers within the vessel wall were made using polarized light microscopy and the universal stage attachment. Data were plotted and analysed using circular statistics. RESULTS: The IMA, structured like an elastic artery, is dominated by a media with discrete lamellae of wavy collagen and smooth muscle, aligned nearly circumferentially, with a low variability of alignment (mean circular SD 12 degrees). The SV is more variable in its size and structure, characteristically with a narrow circumferential media comprised mostly of collagen which is straightened and highly aligned at arterial pressures (mean circular SD 9 degrees). Circumferential collagen in the vein was often adjacent to longitudinal bundles of smooth muscle and collagen. CONCLUSIONS: The strikingly aligned structure of the SV complements the known high mechanical stiffness of this vessel when at arterial distending pressure. The high fraction of longitudinal muscle, in addition to the circumferential muscle cells in the SV make it vulnerable to any pre-implant surgical preparation, and to the cyclical luminal pressures and longitudinal strains characteristic for epicardial arteries.

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.

Cerebral ischemic events associated with prolapsing mitral valve.

Twelve patients who had no evidence of arteriosclerotic cerebral vascular disease, lacked hypertension or coagulation defect, and had not been receiving contraceptive therapy had recurrent transient cerebral ischemic attacks (TIAs) and partial nonprogressive strokes. All had prolapsing mitral valve proved by angiocardiography. The average age was 38 years, compared with 62 years in a larger series of patients with TIA associated with arteriosclerosis. We propose that the ischemic events are related to emboli emanating from the abnormal mitral valve with or without an associated paroxysmal cardiac arrhythmia.

Cardiac and arterial lesions in carotid transient ischemic attacks.

Two hundred fifty consecutive patients with carotid transient ischemic attacks (TIAs) and no previous stroke were assessed with cerebral angiography (95%), two-dimensional echocardiography (86%), electrocardiography (100%), and Holter monitoring (99 selected patients). Angiography disclosed a lesion appropriate to the TIAs in 84%. Lesions also occurred in the asymptomatic carotid artery, but stenosis of more than 75% of the lumen diameter and ulcers were significantly more frequent on the symptomatic side. Twenty-three percent of the patients had a potential source of emboli from the heart, usually in the context of symptomatic heart disease. Among the 205 patients who underwent full angiographic and cardiac investigations, 6% had an isolated potential cardiac source of emboli and 19% had a potential cardiac source of emboli associated with appropriate carotid disease. The search for a potential cardiac source of emboli is strongly indicated in patients with carotid TIAs and known heart disease. In the patients with no history of heart disease, the yield of this search is low, but our results suggest that at least 14 of such patients have an undetected potential cardiac source of emboli. Cardiac and arterial lesions commonly coexist in carotid TIAs.

Clinical predictors of cardiac and arterial lesions in carotid transient ischemic attacks.

In 205 patients with carotid transient ischemic attacks (TIAs) who underwent full angiographic and cardiac investigations, we found that ipsilateral carotid bruit and triggering of TIA by exertion or standing up suggested a carotid lesion. Angina pectoris or palpitations at onset of TIA suggested a cardiac source of emboli. Patients with only one TIA, multiple identical attacks, progression of symptoms over minutes, and appropriate infarct on computed tomograms (28%) were likely to show a potential arterial or cardiac cause for the TIAs in subsequent investigations. Transient monocular blindness correlated with carotid occlusion, but the severity of the carotid lesion did not influence the duration and repetition of attacks. Transient ischemic attacks in multiple territories, identical attacks, and multiple or silent infarcts on computed tomograms occurred equally in the patients with arterial lesions and those with cardiac lesions. These findings suggest that only a few, albeit important, conclusions regarding etiology can be drawn from the clinical characteristics of TIAs.

Mitral valve prolapse and cerebral ischemic events in young patients.

We evaluated 32 young patients with mitral valve prolapse (MVP) as the only recognized cause of cerebral ischemia. The mean follow-up from the time of the first ischemic event was 8 years. At the time of the follow-up evaluation, 24 patients (75%) had a normal cardiac examination, and 4 had midsystolic clicks; only 1 had the characteristic click-murmur. In 75%, the first ischemic event was stroke. When MVP-associated ischemia was recognized, 44% had recurrent ischemic events. In the mean 4-year period between diagnosis of MVP-associated ischemia and follow-up, 16% had recurrent ischemic events, but none had a new persistent deficit from these events. At the time of follow-up, 63% of patients were taking platelet antiaggregants or anticoagulants. These data suggest a good prognosis for recurrent ischemic events after diagnosis of MVP-associated ischemia.

Stress myocardial imaging in mitral leaflet prolapse syndrome.

Mitral leaflet prolapse syndrome has been associated with anginal chest pain, atypical chest pain, electrocardiographic abnormalities and positive stress electrocardiograms. These features overlap those of ischemic heart disease. Furthermore, coronary artery disease is frequently associated with mitral leaflet prolapse. This study evaluated the usefulness of stress myocardial scintigraphy in distinguishing these two disorders. Thirty-two patients with an angiographic diagnosis of mitral leaflet prolapse were studied. Of the 22 patients (8 men and 14 women, mean age 48 years) with a normal coronary arteriogram, 5 had "typical" angina pectoris, 6 had resting electrocardiographic abnormalities and 6 had a positive stress electrocardiogram; all 22 patients had a normal stress myocardial scintigram. Of the 10 patients (7 men and 3 women, mean age 55 years) with at least 70 percent stenosis of one coronary artery, 6 had "typical" angina pectoris, 1 had resting electrocardiographic abnormalities and 7 had a positive stress electrocardiogram. Nine of these 10 patients had one or more demonstrable perfusion defects on stress myocardial scintigrams. It is concluded that mitral leaflet prolapse syndrome is not associated with regional myocardial ischemia as demonstrated with stress scintigraphy, and that stress scintigraphy, a noninvasive technique, is useful in distinguishing the mitral prolapse syndrome from mitral prolapse associated with coronary artery disease.

Transcutaneous continuous wave Doppler ultrasound in the diagnosis of left atrial myxoma.

Six cases of left atrial myxoma were studied using transcutaneous, continuous-wave, low-frequency Doppler ultrasound. The recordings, made from the suprasternal notch and the cardiac apex, showed characteristic deflections that coincided with tumor motion and provided a useful ancillary technique for noninvasively confirming the echocardiographic diagnosis of atrial myxoma.

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.

Aortic valve cusp vessel density: relationship with tissue thickness.

OBJECTIVES: The presence of a microvasculature within aortic cusps implies that tissue oxygen requirements exceed the amount deliverable by diffusion from the tissue surfaces alone. For the design of a successful tissue-engineered valve replacement, the effect of diffusion distance (tissue thickness) on oxygen delivery must be considered. We therefore examined in normal aortic valve cusps the relationship between the presence of microvessels and the tissue thickness. METHODS: Thirty porcine aortic valve cusps were excised and examined after cusp microvessels were pressure filled with a carbon particle solution. Cusp images were captured for stereographic vessel density analysis, and cusp thickness was determined with a radiographic technique. Histologic cross-sections were evaluated to determine vessel depth from the cusp surface. RESULTS: Cusp basal regions measured 0.69 to 0.86 mm in thickness, significantly thicker (P =.001) than the rest of the cusp, which measured 0.36 to 0.48 mm. In general a vascular bed was present when cusp thickness exceeded 0.5 mm, with a median value of 5.16 vessels/mm(3). CONCLUSIONS: From published values of arterial wall oxygen consumption and diffusivity, we predicted that the probable maximum oxygen diffusion distance for valve tissue would be about 0.2 mm. This was consistent with our physical findings, which implies that central tissue anoxia is avoided by the capillary bed. An avascular tissue-engineered valve metabolically similar to an aortic valve should therefore not exceed a thickness of approximately 0.40 mm.

Marked paradoxical septal motion associated with an early diastolic heart sound.

The findings in four patients with marked paradoxical septal motion and an unusual early diastolic sound are presented. Each patient had markedly abnormal right ventricular function. Phonocardiograms, apexcardiograms, and echocardiograms showed the sound to precede the expected timing of a third heart sound and to coincide with the peak leftward displacement of the paradoxically moving septum in early diastole. In two patients the sound was also accompanied by third and fourth heart sounds, revealing a total of five heart sounds on phonocardiographic studies. The sound appeared to originate from the sudden deceleration of the septal mass as it moved leftward during ventricular relaxation.

A simple echocardiographic method to detect atrioventricular dissociation. A useful aid in the differential diagnosis of regular tachycardia with wide QRS complexes.

Regular tachycardia with wide QRS complexes may be difficult to classify as supraventricular (SVT) or ventricular (VT) without electrophysiologic studies. We felt that hemodynamic differences between VT and SVT should allow their distinction by echocardiography. In this study, we utilized high speed M-mode echocardiography in the usual projections in eight patients during tachycardia. Two patients had spontaneous VT and three had spontaneous SVT with aberrant conduction. The remaining three were patients in sinus rhythm undergoing electrophysiologic studies in whom right ventricular (RV) pacing (induced VT), right atrial (RA) pacing (induced SVT) and sequential RV-RA pacing (induced VT with 1:1 retrograde conduction) were carried out. The echocardiographic parameters studied included: left ventricular internal dimensions, time during which the mitral valve remained open (MVOT), left ventricular ejection time (LVET) and pre-ejection period. We measured 20 consecutive beats and for each parameter defined its variability. During A-V dissociation (VT, RV pacing) there was always a striking beat-to-beat variability in the values of MVOT (68 percent to 129 percent) and in LVET (41 percent to 175 percent). In contrast, during A-V association (SVT, sequential RV-RA pacing, sinus rhythm) the maximal variability of MVOT and LVET was 22 percent and 12 percent, respectively. Variability during A-V dissociation could be explained by asynchronous timing of atrial systole. We conclude that echocardiography can readily identify atrioventricular dissociation, a feature heavily in favor of a diagnosis of VT.

Mechanical suitability of glycerol-preserved human dura mater for construction of prosthetic cardiac valves.

We have examined the tensile viscoelastic properties of fresh and glycerol-preserved human dura mater, and correlated the results with structural information from the scanning electron microscope. The interwoven laminar structure of dura produces rather high flexural stiffness, while the crossed-fibrillar laminae produce planar mechanical isotropy. Glycerol storage shifts the stress-strain curve to lower strain, reduces stress relaxation and creep, and lowers the ultimate tensile strength and strain at fracture. These changes may be due to glyceraldehyde crosslinking, or to increased interfibrillar friction. The latter hypothesis suggests that glycerol storage may reduce the fatigue lifetime of the tissue.

Enhanced detection of cardiac myocyte damage by polarized light microscopy. Use in a model of coxsackievirus B3-induced myocarditis.

Although accurate detection of cardiac muscle damage is critical in the diagnosis of acute myocarditis or acute cellular rejection in both clinical and experimental settings, the histologic evaluation is frequently uncertain without specialized stains. In a study of adult male A/J mice infected with 2x10(5) plaque-forming units of myocarditic coxsackievirus B3, cardiac muscle injury causing myofibrillar disruption was detected as a loss of muscle birefringence by polarized light microscopy. The technique was corroborated by comparison with Masson's trichrome stain and was helpful for histologic examination especially at the early preinflammatory stages of lesion development or in fringe territories of focal lesions. Polarized light microscopy is thus an available means to enhance the histologic determination of cardiac myocyte damage and has specific advantage in an absence of specialized stains.

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.

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.

Effect of altered hydration on the internal shear properties of porcine aortic valve cusps.

BACKGROUND: Dehydration of tissue due to glutaraldehyde fixation has been reported and was examined in this study of porcine aortic valve cusps. The effect of altered hydration on cusp internal shear properties was also examined. METHODS: Hydration level was assessed by wet mass measurement of cusps stored in solutions for times up to 1000 minutes. Solutions used in this study included Hanks solution, porcine blood, 0.5% glutaraldehyde, and several dextran solutions. Shear testing was performed on physiologically hydrated, superhydrated, and dehydrated cusps. RESULTS: There was very little difference between the physiologic and superhydrated leaflets; however, dehydration caused significant stiffening with increased hysteresis and stress relaxation. CONCLUSIONS: Glutaraldehyde has been shown to increase shear stiffness of valve cusps. Tissue dehydration also increased shear stiffness but increased stress relaxation and hysteresis, which was contrary to observations reported after glutaraldehyde fixation. The significant effect of dehydration on cusp mechanical properties does not account for the effects observed after glutaraldehyde fixation, but it demonstrates that hydration level is an important factor that affects internal shear properties of valve cusps.

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.

Laser-induced fluorescence (LIF) recognition of the structural composition of porcine heart valves.

Reconstruction and replacement of heart valves with grafts fro pig tissue is a common procedure. However, bioprosthetic valves wear out in a shorter time span than mechanical valves. Bioprosthetic valve structure may contribute to degenerative changes that lead to valve failure. There is, at present, no method to examine the structure of a tissue valve prior to implant. Laser-induced fluorescence (LIF) of natural fluorophores is an elegant method developed for the detection of tumors, dermal lesions and atherosclerosis. We have studied LIF as a potential diagnostic technique for analysis of valvular tissue. Using excimer laser excitation, we examined natural fluorescence recorded from porcine aortic, mitral and pulmonary valves. All three valve outflow surface tissue layers are less fluorescent at 390-450 nm than the inflow layers. Immunohistochemical analysis of collagen I and elastin content in inflow and outflow surface layers of all three valves correlated well with LIF intensities and dI/d lambda values at selected wavelengths. In conclusion, the differences observed in emitted LIF from valve surface layers are found to correlate well with diversity in the structural protein content. The LIF spectroscopic measurements may provide an appropriate tool for examination of tissue valve structure prior to use for implantation.