Ultrasonic characterization of human cancellous bone in vitro using three different apparent backscatter parameters in the frequency range 0.6-15.0 mhz.

Ultrasonic techniques based on measurements of apparent backscatter may provide a useful means for diagnosing bone diseases such as osteoporosis. The term "apparent" means that the backscattered signals are not compensated for the frequency-dependent effects of attenuation and diffraction. We performed in vitro apparent backscatter measurements on 23 specimens of human cancellous bone prepared from the left and right femoral heads of seven donors. A mechanical scanning system was used to obtain backscattered signals from each specimen at several sites. Scans were performed using five different ultrasonic transducers with center frequencies of 1, 2.25, 5, 7.5, and 10 MHz. The -6 dB bandwidths of these transducers covered a frequency range of 0.6-15.0 MHz. The backscattered signals were analyzed to determine three ultrasonic parameters: apparent integrated backscatter (AIB), frequency slope of apparent backscatter (FSAB), and time slope of apparent backscatter (TSAB). Linear regression analysis was used to examine the correlation of these ultrasonic parameters with five measured physical characteristics of the specimens: mass density, X-ray bone mineral density, Young's modulus, yield strength, and ultimate strength. A total of 75 such correlations were examined (3 ultrasonic parameters x 5 specimen characteristics x 5 transducers). Good correlations were observed for AIB using the 5 MHz (r = 0.70 - 0.89) and 7.5 MHz (r = 0.75-0.93) transducers; for FSAB using the 2.25 MHz (r = 0.70 - 0.88), 5 MHz (r = 0.79 - 0.94), and 7.5 MHz (r = 0.80 - 0.92) transducers; and for TSAB using the 5 MHz (r = 0.68 - 0.89), 7.5 MHz (r = 0.75 - 0.89), and 10 MHz (r = 0.75 - 0.92) transducers.

Ultrasonic characterization of cancellous bone using apparent integrated backscatter.

Apparent integrated backscatter (AIB) is a measure of the frequency-averaged (integrated) backscattered power contained in some portion of a backscattered ultrasonic signal. AIB has been used extensively to study soft tissues, but its usefulness as a tissue characterization technique for cancellous bone has not been demonstrated. To address this, we performed measurements on 17 specimens of cancellous bone over two different frequency ranges using a 1 MHz and 5 MHz broadband ultrasonic transducer. Specimens were obtained from bovine tibiae and prepared in the shape of cubes (15 mm side length) with faces oriented along transverse (anterior, posterior, medial and lateral) and longitudinal (superior and inferior) principal anatomic directions. A mechanical scanning system was used to acquire multiple backscatter signals from each direction for each cube. AIB demonstrated highly significant linear correlations with bone mineral density (BMD) for both the transverse (R2 = 0.817) and longitudinal (R2 = 0.488) directions using the 5 MHz transducer. In contrast, the correlations with density were much weaker for the 1 MHz transducer (R2 = 0.007 transverse, R2 = 0.228 longitudinal). In all cases where a significant correlation was observed, AIB was found to decrease with increasing BMD.

Effect of collagen and mineral content on the high-frequency ultrasonic properties of human cancellous bone.

The technology surrounding ultrasonic bone assessment is evolving rapidly as investigators explore the utility of new ultrasonic parameters and different ultrasonic frequencies. This study had three main goals. The first was to perform in vitro measurements of the speed of sound (SOS) and normalized broadband ultrasonic attenuation (nBUA) in specimens of normal human cancellous bone using a 2.25 MHz broadband measurement system. The second was to explore the utility of a backscatter-based parameter called apparent integrated backscatter (AIB). The third goal was to investigate the roles that collagen and mineral content play in affecting each of these three ultrasonic parameters. This was accomplished by chemically treating the specimens to remove one or the other of these two important constituents of bone. Our results showed that in most cases SOS and nBUA correlated well (p < 0.05) with bone mineral density (BMD) as measured by quantitative computed tomography (QCT). In contrast, AIB did not correlate strongly with BMD. When the specimens were demineralized, decreases were produced in SOS (19-39%) and nBUA (44-58%). Changes produced in AIB were not significant except along the superoinferior direction, in which a 12% decrease was measured. When the specimens were decollagenized, decreases were produced in SOS (10-12%). In contrast, increases were produced in both nBUA (35-77%) and AIB (14-15%). From this study we conclude that high-frequency ultrasonic measurements may yield useful information about the content and organization of both collagen and mineral in cancellous bone.

Mechanisms by which AC leakage currents cause complete hemodynamic collapse without inducing fibrillation.

INTRODUCTION: The first study of weak alternating current (AC) stimulation in closed chest humans showed that complete hemodynamic collapse can occur below the threshold for inducing ventricular fibrillation (VF), a heretofore unknown danger to patients. This article, and the accompanying simulation article, explore the mechanisms responsible for the collapse. METHODS AND RESULTS: A quadripolar pacing catheter was placed in the right ventricle (RV) of six dogs. The tip of the catheter (17 mm2) carried 5 seconds of AC stimulation ranging from 10 to 160 Hz and 10 to 1,000 microA. The lead II body surface ECG, femoral artery pressure, and a bipole from the proximal pair of electrodes on the RV catheter were recorded 2 seconds before, during, and 2 seconds after stimulation. Based on the blood pressure, every episode was categorized as VF, COLLAPSE without VF, extrasystolic without COLLAPSE (EFFECT), or having caused no effect (NSR). The electrical activation interval (interspike interval [ISI]) from the RV bipole was compared with the mechanical activation interval, determined from M-mode ultrasound. COLLAPSE is associated with a short ISI (NSR = 408+/-110 msec; EFFECT = 305+/-113 msec; COLLAPSE = 179+/-25 msec; P < 0.001) with a high degree of regularity (P < 0.001): coefficient of variation of ISI for COLLAPSE (0.038+/-0.069) versus VF (0.389+/-0.222), EFFECT (0.420+/-0.241), and NSR (0.016+/-0.048). Electrical activation and mechanical activation rates occurred at integer multiples of the AC stimulation period. CONCLUSION: COLLAPSE (86+/-37 microA; minimum 50 microA in two animals) occurs below the VF threshold (108+/-28 microA) by causing rapid, regular excitation.

Defibrillation and the geometry of the heart: a novel measurement with implications for defibrillation mechanisms.

We present a novel measurement for studying defibrillation mechanisms: the time course of changes in the size of the left ventricular (LV) cavity within 500 ms following defibrillation. Mechanical changes can be linked to electrical mechanisms via an understanding of excitation-contraction coupling. Eight mongrel dogs were internally defibrillated 5-50 seconds (including backup shocks) after the onset of 20 ventricular fibrillation (VF) episodes per animal. Two dimensional, short axis, LV cavity, ultrasound images were recorded at 30 frames per second just prior to inducing VF, during defibrillation and following the shock. Each frame was individually analysed to yield the LV cavity area as a function of time. Defibrillation shocks were followed by a highly reproducible phenomenon: (1) a dramatic and rapid increase in LV area, (2) a more or less prominent LV area plateau and (3) a decrease in the LV area. The peak fractional area increase ranged from 1.65 to 4.64 times larger than the baseline (LV area just prior to defibrillation), averaging 2.18 +/- 0.686. Successful shocks took significantly longer (p < 0.01) to return to 1.3 times the baseline (407 +/- 209 ms) than unsuccessful shocks (296 +/- 130 ms). Extrapolating to electrical mechanisms, our novel measurement demonstrates that defibrillation causes immediate relaxation and therefore suggests a significant role for deexcitation in defibrillation.

Ultrasonic characterization of the curing process of hydroxyapatite-modified bone cement.

Ultrasonic parameters such as velocity of sound and broad-band ultrasonic attenuation (BUA) are sensitive to changes in the viscoelastic properties of a material. Bone cement undergoes changes is these properties as it cures. By monitoring the propagation of ultrasonic pulses through a sample of curing bone cement, the curing reaction of polymethylmethacrylate-based (PMMA) bone cement was investigated for hydroxyapatite (HA) concentrations of 0, 10, and 30% (by weight). As the material hardens, the velocity of sound increases by 70%. BUA shows a large peak at the midpoint of the velocity transition. These data are used to compare the cure time and cure duration for PMMA bone cement mixed with hydroxyapatite particles. Measurements of the final sound velocity and BUA were also performed to investigate the mechanical properties of the fully cured cement, and to compare to compression testing data. This is the first time the curing process of bone cement has been investigated as a function of hydroxyapatite concentration. Results indicate that the cure time is not significantly affected by the addition of HA particles, and that both velocity of sound and BUA are sensitive to the curing process.

Hemodynamic collapse, geometry, and the rapidly paced upper limit of ventricular vulnerability to fibrillation by T-wave stimulation.

There is an upper limit to the vulnerability (ULV) of the ventricles to fibrillation (VF) induced by T-wave stimuli. Across species, disease states, and pharmacological treatments, the ULV is correlated to the defibrillation threshold (DF50). However, one factor known to increase the ULV far above the DF50 is rapid pacing. In this article we test the hypothesis that this increase is owing to an accompanying hemodynamic collapse or geometric change. In 18 dogs, T-wave stimuli were delivered from transvenous defibrillating electrodes. The T-wave shock strength that induced VF 50% of the time (the ULV50) was measured using a 10-step Bayesian up-down protocol. T-wave stimuli were delivered after 15 paced beats at one of several rates: normal (80% of the R-R interval), rapid (the interval just fast enough to cause hemodynamic collapse), or 10 milliseconds greater than rapid (which did not cause hypotension). We measured the geometry of the left ventricle at the moment of T-wave stimulation using linear ultrasound. Rapid pacing significantly increased the ULV50 above the normal rate ULV (507 +/- 62.9 vs 379 +/- 70.6 V, P < .005, n = 18), even in the subset without hemodynamic collapse (505 +/- 84.4 vs 394 +/- 66.5 V, P < .005, n = 6). No significant geometric changes were noted between rapid (19.8 mm) and normal (20.6 mm, n = 6, P < NS) pacing, but QT interval reduction appears to correlate with the ULV50 (QT vs ULV50, r > 0, P < .01). Rapid pacing can dramatically increase the measured ULV50. The most likely cause is a concurrent change in the electrophysiology, eg, QT or APD, of the myocardium. As the only known factor to consistently alter the relationship between ULV and the DF50, rapid pacing offers a unique opportunity for the study of the link between defibrillation and ULV testing.

Anisotropy of Young's modulus of human tibial cortical bone.

The anisotropy of Young's modulus in human cortical bone was determined for all spatial directions by performing coordinate rotations of a 6 by 6 elastic stiffness matrix. The elastic stiffness coefficients were determined experimentally from ultrasonic velocity measurements on 96 samples of normal cortical bone removed from the right tibia of eight human cadavers. The following measured values were used for our analysis: c11 = 19.5 GPa, c22 = 20.1 GPa, c33 = 30.9 GPa, c44 = 5.72 GPa, c55 = 5.17 GPa, c66 = 4.05 GPa, c23 = 12.5 GPa. The remaining coefficients were determined by assuming that the specimens possessed at least an orthorhombic elastic symmetry, and further assuming that c13 = c23 c12 = c11 - 2c66. Our analysis revealed a substantial anisotropy in Young's modulus in the plane containing the long axis of the tibia, with maxima of 20.9 GPa parallel to the long axis, and minima of 11.8 GPa perpendicular to this axis. A less pronounced anisotropy was observed in the plane perpendicular to the long axis of the tibia. To display our results for the full three-dimensional anisotropy of cortical bone, a closed surface was used to represent Young's modulus in all spatial directions.

Low-megahertz ultrasonic properties of bovine cancellous bone.

Ultrasound offers a noninvasive means to detect changes that occur to the density of cancellous bone as a result of degenerative diseases such as osteoporosis. Techniques based on the velocity and frequency dependence of attenuation of ultrasonic pulses propagated through cancellous bone have proven sensitive to bone density. Most previous studies have investigated these two parameters in the frequency range of 0.1-1.0 MHz. The present study had two goals. The first was to measure three ultrasonic parameters: longitudinal mode velocity; broadband ultrasonic attenuation (BUA); and apparent integrated backscatter (AIB), at higher frequencies using a broadband 2.25 MHz measurement system. The second goal was to assess the dependence of these parameters on bone density. Twenty-one specimens of cancellous bone acquired from the proximal end of four bovine tibiae were investigated in this study. The apparent density of the specimens (determined with the bone marrow removed and the specimens thoroughly dry) ranged between 0.3 and 0.9 g/cm(3). Ultrasonic measurements were performed along three mutually perpendicular directions corresponding to the anteroposterior (AP), mediolateral (ML), and superoinferior (SI) axes of the tibia. A linear regression was used to analyze the results of these measurements as a function of apparent density. Velocity demonstrated a highly significant linear increase with density for all three directions (AP: p < 0.001; ML: p < 0.001; SI: p < 0.01). AIB decreased with density in all three directions; however, only the ML and SI directions demonstrated a significant linear correlation (AP: p = n.s.; ML: p < 0.05; SI: p < 0.05). In the frequency range 0.5-1.0 MHz, BUA exhibited a significant linear increase in the AP and ML directions, but not the SI direction (AP: p < 0.05; ML: p < 0.01; SI: p = n.s.). In contrast, in the frequency range 1.0-2.0 MHz, BUA exhibited a highly significant increase with density in the SI direction, but no significant change in the AP and ML directions (AP: p = n.s., ML: p = n.s., SI: p < 0.001).

Ultrasonic determination of the anisotropy of Young's modulus of fixed tendon and fixed myocardium.

The linear elastic properties of a soft tissue exhibiting a unidirectional arrangement of reinforcing fibers may be described in terms of the five independent elastic stiffness coefficients C11, C13, C33, C44, and C66. In previous studies, ultrasonic measurements of these coefficients for formalin fixed specimens of bovine Achilles tendon and normal human myocardium were reported. In the present study these results are used to analyze the anisotropy of Young's modulus of these tissues. For formalin fixed tendon a value of 1.37 GPa is obtained for Young's modulus along the fiber axis of the tissue, and a value of 0.0706 GPa is obtained perpendicular to the fibers. For formalin fixed myocardium, values of 0.101 and 0.0311 GPa parallel and perpendicular to the fibers, respectively, are obtained. Based on the results for the angular dependence of Young's modulus from unidirectional specimens of myocardium, a model is introduced to estimate these features for the more complicated fiber architecture of the left ventricular wall.

Anisotropy of the transverse mode ultrasonic properties of fixed tendon and fixed myocardium.

This study investigates the influence of the fiber-reinforced nature of myocardium and tendon on the propagation of transverse mode ultrasonic waves. Formalin fixed specimens of normal human left ventricular cardiac muscle and bovine Achilles tendon were prepared for this study in such a way that transverse mode ultrasonic waves could be propagated perpendicular to the fiber axis of the tissue with the polarization oriented either parallel or perpendicular to the fiber axis. Measurements of velocity and attenuation were made at 3 MHz to assess the degree of anisotropy in these parameters for both tissues. Formalin fixed tendon exhibited a significant anisotropy whereas formalin fixed myocardium displayed a similar trend of more modest magnitude. Results of these measurements were used to compute two elastic stiffness coefficients for each tissue, yielding c44 = 37.2 MPa and c66 = 18.0 MPa for formalin fixed tendon, and c44 = 8.97 MPa and c66 = 8.45 MPa for formalin fixed myocardium. To validate this approach, additional studies were conducted to measure the transverse mode ultrasonic properties of silicone rubber and motor oil.

Anisotropy of the slope of ultrasonic attenuation in formalin fixed human myocardium.

Clinical implementation of quantitative ultrasonic tissue characterization is likely to require imaging the heart with sound propagating at varying angles relative to the fibers of the heart. Under these circumstances, the variation of the ultrasonic properties of myocardium with the angle of propagation relative to the myofibers may represent a significant source of potential misinterpretation. In the present study, the systematic approach of assessing the impact of anisotropy on quantitative myocardial tissue characterization is extended by reporting results of a recent in vitro study to measure the anisotropy of the slope of ultrasonic attenuation in specimens of formalin fixed human myocardium. Data obtained from regions of remote infarct are presented and compared to data acquired from regions identified to be free of infarct. The slope of attenuation for both regions exhibit a sinusoid-like dependence on angle that is approximately doubled for propagation parallel to the fibers as compared to perpendicular. These results are, in turn, compared to an earlier study from the laboratory that examined the effects of myocardial infarction on ultrasonic attenuation and interstitial collagen content in freshly excised canine hearts. Discussion regarding the analysis and interpretation of measurements of slope of attenuation is presented as well as a discussion of the possible influence of formalin fixation on our results.

Estimation of the elastic stiffness coefficient c13 of fixed tendon and fixed myocardium.

Recent studies from our laboratory have detailed the anisotropy of velocity of quasilongitudinal-mode ultrasonic waves through formalin fixed samples of normal human myocardium and bovine Achilles tendon. Results of these studies were used to determine the elastic stiffness coefficients c33, corresponding to the propagation of longitudinal-mode waves parallel to the fiber axis of the tissue, and c11, corresponding to the propagation of longitudinal-mode waves perpendicular to the fiber axis. For a tissue possessing a unidirectional arrangement of fibers with a random transverse distribution, three additional coefficients, c13, c44, and c12, are needed to describe its linear mechanical properties completely. Direct ultrasonic measurements of these coefficients in solids typically require the propagation of transverse-mode waves through the sample. Such measurements are difficult to perform in soft tissues because transverse-mode ultrasonic waves are very highly attenuated by the tissue. This study therefore employs a technique to estimate c13 based on measurements of the velocity of quasilongitudinal-mode ultrasonic waves for numerous angles of propagation relative to the fiber axis of the tissue. Analysis of data obtained from formalin fixed bovine Achilles tendon and human myocardium yield estimated values for c13 of 3.17 and 2.46 GPa, respectively.

Comparison of the anisotropy of apparent integrated ultrasonic backscatter from fixed human tendon and fixed human myocardium.

The content and organization of collagen in the cardiac interstitium may represent significant determinants of the ultrasonic scattering properties of myocardium. This study was designed to investigate the anisotropic backscattering properties of a fibrous soft tissue exhibiting an ordered arrangement of fibers similar to myocardium, but possessing a substantially greater content of collagen. Human Achilles tendon was chosen for this study because it possesses a simple unidirectional arrangement of fibers and a high content of collagen compared to normal myocardium. Integrated (frequency-averaged) backscatter was measured from ten formalin fixed samples of tendon as a function of insonifying angle relative to the fiber axis of the tissue. The samples were insonified in a water bath using a 5-MHz center frequency piezoelectric transducer. Maximum backscatter occurred for insonification perpendicular to the fibers, and minimum backscatter occurred for insonification parallel to the fibers. The mean peak to nadir variation, or magnitude of anisotropy, of integrated backscatter for the ten formalin fixed samples of tendon was 36.3 dB. This compares to 14.5 dB for formalin fixed human myocardium measured in an earlier study by our laboratory.

Effect of collagen on the anisotropy of quasi-longitudinal mode ultrasonic velocity in fibrous soft tissues: a comparison of fixed tendon and fixed myocardium.

The widespread use of echocardiography has generated considerable interest in the ultrasonic properties of myocardial collagen. This study was designed to investigate the effect of collagen on the propagation of ultrasound by measuring the anisotropy of ultrasonic velocity through formalin fixed specimens of bovine Achilles tendon. Tendon was chosen for this study because it possesses a high content of collagen and a well-defined unidirectional arrangement of fibers. Ultrasonic velocity data were acquired from nine samples of fixed tendon that were each insonified at multiple angles relative to the fibers in 2 degrees increments for a full 360 degrees. Analysis of the data revealed a substantial angular dependence of velocity qualitatively similar to that reported for formalin fixed specimens of normal human myocardium, but approximately 17 times larger in magnitude. Together with measured values of density, these results were used to compute the elastic stiffness coefficients C11 (corresponding to propagation perpendicular to the fibers) and C33 (corresponding to propagation parallel to the fibers) of fixed tendon, yielding 3.08 and 4.51 GPa, respectively.

Detection of unique transmural architecture of human idiopathic cardiomyopathy by ultrasonic tissue characterization.

BACKGROUND: Noninvasive approaches to the evaluation of idiopathic cardiomyopathy are limited. Recent work from our laboratory has used quantitative ultrasound to define the three-dimensional structure of normal human myocardium and the myocardial remodeling associated with infarction. Our goal was to define the role of ultrasonic tissue characterization for detection of specific alterations in the three-dimensional transmural architecture of idiopathic dilated cardiomyopathy. METHODS AND RESULTS: We measured frequency-dependent backscatter from 22 cylindrical biopsy specimens from nine explanted fixed hearts of patients who underwent heart transplantation for idiopathic cardiomyopathy, seven specimens from normal portions, and 12 specimens of infarcted tissue from six explanted fixed human hearts. Consecutive transmural levels from each specimen were insonified with a 5-MHz broadband transducer. The dependence of apparent (uncompensated for attenuation) backscatter, B(f), on frequency (f) was computed from radiofrequency (rf) data as: magnitude of B(f)2 = afn, where n is an index that reflects in part the size of the dominant scatterers in myocardial tissue. Myofiber diameter and percentage fibrosis were determined at each transmural level for each specimen. For cardiomyopathic tissue, the frequency dependence of backscatter (n) increased progressively from epicardial to endocardial (0.02 +/- 0.37 to 1.01 +/- 0.12, p less than 0.05) levels in conjunction with a progressive decrease in myofiber diameter (29.5 +/- 0.9 to 21.4 +/- 0.6 microns, p less than 0.0001). In contrast, in tissue from areas of infarction, the frequency dependence decreased progressively from epicardium to endocardium (0.91 +/- 0.20 to 0.23 +/- 0.21, p less than 0.05) in conjunction with a progressive increase in the percentage of fibrosis (23.5 +/- 9.4% to 54.5 +/- 4.9%, p less than 0.005). Normal tissue exhibited no significant transmural trend for frequency dependence, myofiber diameter, or percentage fibrosis. CONCLUSIONS: These data indicate the presence of a heterogenous transmural distribution of scattering structures associated with human idiopathic cardiomyopathy and myocardial infarction that may be detected by ultrasonic tissue characterization. The divergence of these transmural trends for frequency dependence of backscatter reflects distinct mechanisms of structural heterogeneity for different pathological processes that comprise a transmural gradation of cell size and fibrosis for idiopathic cardiomyopathy and infarction, respectively.