Ultrasound evaluation of mechanical injury of bovine knee articular cartilage under arthroscopic control

A local cartilage injury can trigger development of posttraumatic osteoarthritis (OA). Surgical methods have been developed for repairing cartilage injuries. Objective and sensitive methods are needed for planning an optimal surgery as well as for monitoring the surgical outcome. In this laboratory study, the feasibility of an arthroscopic ultrasound technique for diagnosing cartilage injuries was investigated. In bovine knees (n = 7) articular cartilage in the central patella and femoral sulcus was mechanically degraded with a steel brush modified for use under arthroscopic control. Subsequently, mechanically degraded and intact adjacent tissue was imaged with a high frequency (40 MHz) intravascular ultrasound device operated under arthroscopic guidance. After opening the knee joint, mechanical indentation measurements were also conducted with an arthroscopic device at each predefined anatomical site. Finally, cylindrical osteochondral samples were extracted from the measurement sites and prepared for histological analysis. Quantitative parameters, i.e., reflection coefficient (R), integrated reflection coefficient (IRC), apparent integrated backscattering (AIB), and ultrasound roughness index (URI) were calculated from the ultrasound signals. The reproducibilities (sCV %) of the measurements of ultrasound parameters were variable (3.7% to 26.1%). Reflection and roughness parameters were significantly different between mechanically degraded and adjacent intact tissue (p < 0.05). Surface fibrillation of mechanically degraded tissue could be visualized in ultrasound images. Furthermore, R and IRC correlated significantly with the indentation stiffness. The present results are encouraging; however, further technical development of the arthroscopic ultrasound technique is needed for evaluation of the integrity of human articular cartilage in vivo.     

Microelastic imaging of bone

Several high-frequency ultrasound techniques have been developed during the last decade with the intention of assessing elastic properties of bone at the tissue level. The basic measurement principles can be divided into: 1) measurement of the compressional wave velocity in thin tissue sections; 2) measurement of surface acoustic wave velocities in thick sections; and 3) derivation of the acoustic impedance from the confocal reflection amplitude in thick sections. In this paper, the 3 principles are described with example measurements given in the frequency range from 50 MHz to 1.2 GHz. The measurements were made with 2 microscopes operating in the pulse-echo mode, either with frequencies up to 200 MHz and time-resolved detection or between 100 MHz and 2 GHz and amplitude detection. The methods are compared and their application potentials and limitations are discussed with respect to the hierarchical structure of cortical bone. Mapping of the confocal reflection amplitude has superior capabilities for deriving quantitative elastic and structural parameters in the heterogeneous bone material. Even at low frequencies (50 MHz), the mineralized tissue matrix can be separated from the larger pores (Haversian canals), and the elastic coefficient in the probing direction can be measured in 2 dimensions. Depending on the type of sample surface preparation (flat or cylindrically shaped), local distribution of a single elastic coefficient or the average transverse isotropic stiffness tensor can be derived. With frequencies in the GHz range, the lamellar bone structure can be analyzed. However, at one GHz, the acoustic wavelength is still one order of magnitude larger than the individual mineralized collagen fibrils. Although the thickness of a lamellar unit can easily be assessed from the acoustic image, the derivation of the anisotropic elastic properties of the mineralized collagen fibrils as well as the detailed structure of a lamella can only be accomplished with further model assumptions.     

超声弹性显微镜成像系统开发与应用的初步研究

传统的超声弹性成像技术一般使用1MHz～10MHz的超声波,这一频段超声波的空间分辨率在毫米量级,它不能满足对生物组织中微细结构(如皮肤层,关节软骨等)的研究。文中介绍一套新开发的超声弹性显微镜成像系统,并将其初步应用于对关节软骨和老鼠表皮的成像研究中。该系统由加压系统和背向散射超声显微镜系统两部分组成,其中超声探头频率为50MHz。通过对关节软骨和老鼠表皮的成像实验表明,该系统可以清晰的对生物组织中微细结构成像,研究它们的机械特性。     

Multifrequency ultrasound transducers for conformal interstitial thermal therapy

Control over the pattern of thermal damage generated by interstitial ultrasound heating applicators can be enhanced by changing the ultrasound frequency during heating. The ability to change transmission frequency from a single transducer through the use of high impedance front layers was investigated in this study. The transmission spectrum of multifrequency transducers was calculated using the KLM equivalent circuit model and verified with experimental measurements on prototype transducers. The addition of a quarter-wavelength thick PZT (unpoled) front layer enabled the transmission of ultrasound at two discrete frequencies, 4.7 and 9.7 MHz, from a transducer with an original resonant frequency of 8.4 MHz. Three frequency transmission at 3.3, 8.4, and 10.8 MHz was possible for a transducer with a half-wavelength thick front layer. Calculations of the predicted thermal lesion size at each transmission frequency indicated that the depth of thermal lesion could be varied by a factor of 1.6 for the quarter-wavelength front layer. Heating experiments performed in excised liver tissue with a dual-frequency applicator confirmed this ability to control the shape of thermal lesions during heating to generate a desired geometry. Practical interstitial Designs that enable the generation of shaped thermal lesions are feasible.     

Reflection from bound microbubbles at high ultrasound frequencies

Targeted contrast agents and ultrasound imaging are now used in combination for the assessment and tracking of biomarkers in animal models in vivo. These applications have triggered interest in the understanding and prediction of the ultrasound echoes from contrast agents attached to cells. This study describes the reflection enhancement due to microbubbles bound on a gelatin surface. The reflection enhancement was measured using ultrasound pulses at high frequency (40 MHz) and low pressure (38 kPa peak-negative pressure) allowing a linear approximation to be applied. The observed reflection coefficient increased with the number of microbubbles, until reaching saturation at 0.9 when the surface coverage fraction was 35%. A multiple scattering model assuming that the targeted microbubbles are confined within an infinitesimally thin layer appeared suitable in predicting the reflection coefficient even at very high surface densities. These results could permit the optimization of the sensitivity of high frequency ultrasound to targeted contrast agents.     

High frequency ultrasound imaging with optical arrays

Dynamically focused and steered high frequency ultrasound imaging systems require arrays with fine element spacing, wide bandwidths, and large apertures. However, these characteristics are difficult to achieve at frequencies greater than 30 MHz using conventional array construction methods. Optical schemes offer a solution. Focused laser beams incident on a suitable surface can generate and detect acoustic radiation. Precisely controlling the position and size of the beams defines points of transmission and detection, making it possible for pulse-echo image formation by synthetic aperture methods. An optical detection array was built, relying on a conventional piezoelectric transducer as an ultrasound source. The detection system, with near optimal resolution over a wide depth of field, demonstrates the potential for high frequency array implementation using optical techniques. A possible application is in pathology, where 2-D or 3-D fine resolution pulse-echo imaging can be performed in situ without the need for biopsies.     

Binary code design for high-frequency ultrasound

This paper proposes an approach to designing binary codes suitable for high-frequency applications of coded excitation in medical ultrasound. For a high-frequency ultrasound system, transmitting well-designed binary codes with a low sampling ratio (i.e., the bit rate divided by the transducer center frequency) is a practical way to improve the signal-to-noise ratio (SNR) because the challenge of implementing arbitrary-waveform generators for transmitting nonbinary codes increases with the frequency and the switching speed of square-wave pulsers are limited. One conventional approach designs codes using a base sequence that modulates wideband sequences up to the transducer passband. Because a major portion of codes is excluded as a candidate, codes designed using this approach typically need long compression filters for restoring the axial resolution, and they do not improve the SNR efficiently. In contrast, the approach proposed here searches all the codes that match the transducer passband; hence, the resultant codes exhibit better performance. The technique was tested using a bit rate of 50 MHz and a sampling ratio of 2. For a transducer with an ideal Gaussian frequency response with a center frequency of 25 MHz and a -6 dB bandwidth of 15 MHz, the SNR for the same side-lobe extent was 1 to 6 dB higher for the codes designed using the proposed approach compared with those designed using the conventional approach. When a real transducer response with a center frequency of 26.4 MHz and a one-way -6 dB bandwidth of 20.7 MHz was considered, the codes designed using the proposed approach were superior by 0.5 to 5 dB. Therefore, our approach is better than the conventional approach for designing binary codes for high-frequency ultrasound, with the results indicating that the moderate bit rate of 50 MHz will suffice when the ultrasonic center frequency is 25 MHz.     

Microfabrication of electrode patterns for high-frequency ultrasound transducer arrays

High-frequency ultrasound is needed for medical imaging with high spatial resolution. A key issue in the development of ultrasound imaging arrays to operate at high frequencies (?30 MHz) is the need for photolithographic patterning of array electrodes. To achieve this directly on 1-3 piezocomposite, the material requires not only planar, parallel, and smooth surfaces, but also an epoxy composite filler that is resistant to chemicals, heat, and vacuum. This paper reports, first, on the surface finishing of 1-3 piezocomposite materials by lapping and polishing. Excellent surface flatness has been obtained, with an average surface roughness of materials as low as 3 nm and step heights between ceramic/polymer of ～80 nm. Subsequently, high-frequency array elements were patterned directly on top of these surfaces using a photolithography process. A 30-MHz linear array electrode pattern with 50-μm element pitch has been patterned on the lapped and polished surface of a high-frequency 1-3 piezocomposite. Excellent electrode edge definition and electrical contact to the composite were obtained. The composite has been lapped to a final thickness of ～55 μm. Good adhesion of electrodes on the piezocomposite has been achieved and electrical impedance measurements have demonstrated their basic functionality. The array was then packaged, and acoustic pulse-echo measurements were performed. These results demonstrate that direct patterning of electrodes by photolithography on 1-3 piezocomposite is feasible for fabrication of high-frequency ultrasound arrays. Furthermore, this method is more conducive to mass production than other reported array fabrication techniques.     

实时超声膨胀测量系统在关节软骨中的实验研究

关节软骨是覆盖关节表面的一层承载生物重量的组织。关节软骨在正常状态下，软骨蛋白多糖的弹性和胶原纤维的张力保持平衡。这种平衡的微小变化会引起关节软骨的退化。关节软骨特别是其表层区域的膨胀是和骨关节炎的退化相联系的，定量的测量关节软骨的膨胀可以表征骨关节炎的退化变化。本文的主要目的是介绍一种新的实时超声膨胀测量系统，并把该系统应用到关节软骨的研究中。该系统使用50MHz的超声来实时动态观测用胰岛素和0．15M生理盐水溶液分别浸泡处理的牛膝盖关节软骨的非均匀瞬时深度依赖膨胀行为。实验表明，实时超声检测的方法为关节软骨的退化研究提供了独特的工具。这项技术结合关节内窥镜检查，具有潜在的早期诊断体内关节退化的价值。     

A 100-200 MHz ultrasound biomicroscope

The development of higher frequency ultrasound imaging systems affords a unique opportunity to visualize living tissue at the microscopic level. This work was undertaken to assess the potential of ultrasound imaging in vivo using the 100-200 MHz range. Spherically focused lithium niobate transducers were fabricated. The properties of a 200 MHz center frequency device are described in detail. This transducer showed good sensitivity with an insertion loss of 18 dB at 200 MHz. Resolution of 14 /spl mu/m in the lateral direction and 12 /spl mu/m in the axial direction was achieved with f/1.14 focusing. A linear mechanical scan system and a scan converter were used to generate B-scan images at a frame rate up to 12 frames per second. System performance in B-mode imaging is limited by frequency dependent attenuation in tissues. An alternative technique, zone-focus image collection, was investigated to extend depth of field. Images of coronary arteries, the eye, and skin are presented along with some preliminary correlations with histology. These results demonstrate the feasibility of ultrasound biomicroscopy In the 100-200 MHz range. Further development of ultrasound backscatter imaging at frequencies up to and above 200 MHz will contribute valuable information about tissue microstructure.     

Improvements in optical generation of high-frequency ultrasound

The thermoelastic effect in a black polydimethylsiloxane (PDMS) film is used to produce high-frequency, high-intensity ultrasound for applications in water and soft tissue. We show that the optoacoustic transduction efficiency is improved by about 10 dB by decreasing the thickness of the black PDMS film from 25 microm to 11 microm. The center frequency of the generated ultrasound is 60 MHz, with a -6 dB bandwidth of 80%. When a 5 ns laser pulse with energy of 50 microJ is delivered to a spot size of 25 microm, the acoustic pressure 10 mm away from the film surface is about 800 kPa. Furthermore, we demonstrate that the center frequency and bandwidth of the generated ultrasound is mainly determined by the temporal profile of the input optical pulse, and it has the potential to be easily improved to above 100 MHz.     

Transduction mechanisms of the Fabry-Perot polymer film sensing concept for wideband ultrasound detection

The transduction mechanisms of a wideband (30 MHz) contact ultrasound sensor based upon the use of a thin polymer film acting as a Fabry-Perot interferometer have been investigated. Polyethylene terepthalate (PET) sensing elements, illuminated by the free-space collimated output of a wavelength-tunable DBR laser diode, have been used to study the sensor transfer function, sensitivity, the effect of water absorption, and frequency response characteristics. Acoustic performance was evaluated by comparing the sensor output with that of a calibrated PVDF membrane hydrophone using laser-generated acoustic transients as a source of broadband ultrasound. An ultrasonic acoustic phase sensitivity of 0.1 rad/MPa, a linear operating range to 5 MPa, and a noise-equivalent-pressure of 20 kPa over a 25 MHz measurement bandwidth were obtained using a water-backed 50 mum PET sensing film. A model of frequency response that incorporates the effect of an adhesive layer between the sensor film and backing material has been developed and validated for different sensing film thicknesses, backing configurations, and adhesive layer thicknesses.     

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.     

Identifying chemical changes in subchondral bone taken from murine knee joints using Raman spectroscopy

Application of Raman spectroscopy to analysis of subchondral bone is described. The effect of cartilage health on subchondral bone has been widely studied using radiological and histological methods; however, there is no method to directly assay mineral components. We present Raman spectra of femur condyles and observe mineral bands that arise from the subchondral bone. In two separate experiments, transgenic mouse models of early-onset osteoarthritis (OA) and lipoatrophy were compared to tissue from wild-type mice. Raman spectroscopy was used to identify chemical changes in the mineral of subchondral bone that may accompany or precede morphological changes that can be observed by histology. The transgenic mice were compared to age-matched wild-type mice. Subtle alterations in the mineral or collagen matrix were observed by Raman spectroscopy using established Raman markers such as the carbonate-to-phosphate ratio, mineral-to-matrix ratio (MTMR), and amide I ratio. The Raman microscope configuration enabled rapid collection of Raman spectra from the mineralized layer that lies under an intact layer of non-mineralized articular cartilage. The effect of the cartilage layer on collection of spectra is discussed. The technique proposed is capable of providing insight into the chemical changes that occur in subchondral bone on a molecular level.     

The regeneration of articular cartilage using a new polymer system

A polymer system based on room temperature polymerising poly (ethylmethacrylate) polymer powder and tetrahydrofurfuryl monomer has been investigated as a biomaterial for encouraging articular cartilage repair. This heterocyclic methacrylate polymer system swells slightly in situ and thus provides a good interface with subchondral bone resulting in mechanical stability with favourable uptake kinetics. Another feature of this polymer system is that it exhibits high water uptake which leads to absorption of the surrounding tissue fluid and matrix proteins, including growth factors; this may encourage the formation of new cartilage. Three weeks after implantation the tissue overgrowth contained cartilage components: chondrocytes, collagen type II, chondroitin 4-sulphate and chondroitin 6-sulphate. In addition numerous chondrocyte clones were observed at the edge of the defect and in the newly repaired tissue. By six weeks a superficial articulating surface was continuous with the normal articular cartilage with underlying tissue which showed some evidence of endochondral ossification. By nine weeks the surface covering of new cartilage had a widened and an irregular zone of calcified cartilage with thickened subchondral bone was present. At eight months the resurfaced cartilage remained intact above a remodelled subchondral bone end plate.     

Dynamic response of a frequency measuring system

A frequency measurement system using a double-buffered method is designed for PC/AT compatible computers. The extremely wide range of measured frequencies (from 1.67 mHz to 7 MHz) is independent of a wide range of sampling periods (from 100 μs to 300 s). The response time to an instantaneously increased frequency from 1.67 mHz to 7 MHz is less than two sampling periods. The menu-driven software graphically presents frequency versus time to analyze fast frequency change using rapid sampling, or to measure the average frequency during long sampling periods     

Dual-pulse frequency compounded superharmonic imaging

Tissue second-harmonic imaging is currently the default mode in commercial diagnostic ultrasound systems. A new modality, superharmonic imaging (SHI), combines the third through fifth harmonics originating from nonlinear wave propagation through tissue. SHI could further improve the resolution and quality of echographic images. The superharmonics have gaps between the harmonics because the transducer has a limited bandwidth of about 70% to 80%. This causes ghost reflection artifacts in the superharmonic echo image. In this work, a new dual-pulse frequency compounding (DPFC) method to eliminate these artifacts is introduced. In the DPFC SHI method, each trace is constructed by summing two firings with slightly different center frequencies. The feasibility of the method was established using a single-element transducer. Its acoustic field was modeled in KZK simulations and compared with the corresponding measurements obtained with a hydrophone apparatus. Subsequently, the method was implemented on and optimized for a setup consisting of an interleaved phased-array transducer (44 elements at 1 MHz and 44 elements at 3.7 MHz, optimized for echocardiography) and a programmable ultrasound system. DPFC SHI effectively suppresses the ghost reflection artifacts associated with imaging using multiple harmonics. Moreover, compared with the single-pulse third harmonic, DPFC SHI improved the axial resolution by 3.1 and 1.6 times at the -6-dB and -20-dB levels, respectively. Hence, DPFC offers the possibility of generating harmonic images of a higher quality at a cost of a moderate frame rate reduction.     

强度超声对斑马鱼胚胎发育的影响

超声波的生物学效应一直深受人们关注,低强度超声对细胞的作用机理也不断地被人们所了解。为了研究超声对斑马鱼胚胎发育的影响,通过声学共振理论以及有限元仿真得到圆球形细胞及椭球形细胞的共振频率,并设计了一种低强度超声装置对斑马鱼胚胎进行刺激。通过正交试验设计研究了超声刺激频率、刺激时间和信号激励电压对鱼卵孵化指数的影响,实验结果表明：在频率为1.26 MHz,刺激时间为30 min,信号激励电压为3 Vpp条件下鱼卵孵化指数明显高于对照组,且最佳刺激频率与仿真计算的结果基本一致。由此可见,本研究前期的理论推导及仿真计算结果可信,也证明了适当条件的超声刺激可以加速斑马鱼胚胎细胞的生长。     

Blood noise reduction in intravascular ultrasound imaging

Scattering from red blood cells (blood noise) increases significantly as the ultrasound frequency is increased above 10 MHz. This reduces the contrast between the vessel wall and the lumen in intravascular ultrasound imaging which makes it difficult to localize the vessel wall and plaque. A blood noise filter based on beam tilting and digital lateral low pass filtering is described. Beam tilting introduces a Doppler shift from blood which results in a frequency separation of the vessel wall signal and the blood noise. The performance of the filter is investigated by simulations and by in vitro experiments. The filter is found to be effective for blood velocities exceeding approximately 50 cm s^-1 at a 20 MHz ultrasound frequency with a beam tilt angle of 10 degrees and a frame rate of 15 f.p.s. By increasing the system frequency to 40 MHz, increase the beam tilt angle to 15 degrees and reduce the frame rate to 10 f.p.s., the filter is effective for blood velocities below 10 cm s^-1     

Two-dimensional localization with a single diffuse ultrasound field excitation

Traditional ultrasound imaging methods rely on the bandwidth and center frequency of transduction to achieve axial and radial image resolution, respectively. In this study, a new modality for spatially localizing scattering targets in a two-dimensional field is presented. In this method, the bandwidth of field excitation is high, and the center frequency is lowered such that the corresponding wavelengths are substantially larger than the target profiles. Furthermore, full two-dimensional field measurements are obtained with single send-receive sequences, demonstrating a substantial simplification of the traditional scanning techniques. Field reconstruction is based on temporal-spectral cross-correlations between measured backscatter data and a library of region of interest (ROI) backscatter data measured a priori. The transducer design is based upon a wedge-shaped geometry, which was shown to yield spatially frequency-separated bandwidths of up to 156% with center frequencies of 1.38 MHz. Initial results with these send-and-receive transducer parameters and cylindrical reflection targets in a 10-mm x 10-mm ROI demonstrate two-dimensional target localization to within 0.5 mm. Spatial localization of point scatterers is demonstrated for single and multiple scattering sites.