This study demonstrates for the first time the development of engineered tissues based on anatomic geometries derived from widely used medical imaging modalities such as computed tomography (CT) and magnetic resonance imaging (MRI). Computer-aided design and tissue injection molding techniques have demonstrated the ability to generate living implants of complex geometry. Due to its complex geometry, the meniscus of the knee was used as an example of this technique's capabilities. MRI and microcomputed tomography (microCT) were used to design custom-printed molds that enabled the generation of anatomically shaped constructs that retained shape throughout 8 weeks of culture. Engineered constructs showed progressive tissue formation indicated by increases in extracellular matrix content and mechanical properties. The paradigm of interfacing tissue injection molding technology can be applied to other medical imaging techniques that render 3D models of anatomy, demonstrating the potential to apply the current technique to engineering of many tissues and organs.
Objectives The purpose of this study was to determine whether the physical properties and vascular environment of active myofascial trigger points associated with acute spontaneous cervical pain, asymptomatic latent trigger points, and palpably normal muscle differ in terms of the trigger point area, pulsatility index, and resistivity index, as measured by sonoelastography and Doppler imaging. Methods Sonoelastography was performed with an external 92-Hz vibration in the upper trapezius muscles in patients with acute cervical pain and at least 1 palpable trigger point (n = 44). The area of reduced vibration amplitude was measured as an estimate of the size of the stiff myofascial trigger points. Patients also underwent triplex Doppler imaging of the same region to analyze blood flow waveforms and calculate the pulsatility index of blood flow in vessels at or near the trigger points. Results On sonoelastography, active sites (spontaneously painful with palpable myofascial trigger points) had larger trigger points (mean ± SD, 0.57 ± 0.20 cm2) compared to latent sites (palpable trigger points painful on palpation; 0.36 ± 0.16 cm2) and palpably normal sites (0.17 ± 0.22 cm2; P < .01). Analysis of receiver operating characteristic curves showed that area measurements could robustly distinguish between active, latent, and normal sites (areas under the curve, 0.9 for active versus latent, 0.8 for active versus normal, and 0.8 for latent versus normal, respectively). Doppler spectral waveform data showed that vessels near active sites had a significantly higher pulsatility index (median, 8.3) compared to normal sites (median, 3.0; P < .05). Conclusions The results presented in this study show that myofascial trigger points may be classified by area using sonoelastography. Furthermore, monitoring the trigger point area and pulsatility index may be useful in evaluating the natural history of myofascial pain syndrome.
Objectives Our objectives were to develop a new, efficient, and easy-to-administer approach to ultrasound elastography and assess its ability to provide quantitative characterization of viscoelastic properties of skeletal muscle in an outpatient clinical environment. We sought to show its validity and clinical utility in assessing myofascial trigger points, which are associated with myofascial pain syndrome. Methods Ultrasound imaging was performed while the muscle was externally vibrated at frequencies in the range of 60 to 200 Hz using a handheld vibrator. The spatial gradient of the vibration phase yielded the shear wave speed, which is related to the viscoelastic properties of tissue. The method was validated using a calibrated experimental phantom, the biceps brachii muscle in healthy volunteers (n = 6), and the upper trapezius muscle in symptomatic patients with axial neck pain (n = 13) and asymptomatic (pain-free) control participants (n = 9). Results Using the experimental phantom, our method was able to quantitatively measure the shear moduli with error rates of less than 20%. The mean shear modulus ± SD in the normal biceps brachii measured 12.5 ± 3.4 kPa, within the range of published values using more sophisticated methods. Shear wave speeds in active myofascial trigger points and the surrounding muscle tissue were significantly higher than those in normal tissue at high frequency excitations (>100 Hz; P < .05). Conclusions Off-the-shelf office-based equipment can be used to quantitatively characterize skeletal muscle viscoelastic properties with estimates comparable to those using more sophisticated methods. Our preliminary results using this method indicate that patients with spontaneous neck pain and symptomatic myofascial trigger points have increased tissue heterogeneity at the trigger point site and the surrounding muscle tissue.
This study investigated the hypothesis that controlled media mixing will enhance tissue formation and increase mechanical properties of anatomically-shaped tissue engineered menisci. Bovine meniscal fibrochondrocytes were seeded in 2% w/v alginate, cross-linked with 0.02 g/mL CaSO4, and injected into molds of menisci. Engineered menisci were incubated for up to 6 weeks. A mixing media bioreactor was designed to ensure proper mixing of culture medium while protecting constructs from the spinning impeller. Impeller speeds were calibrated to produce Reynolds number (Re) of 0.5, 2.9, 5.8, 10.2, and 21.8. Constructs were divided an tested in confined compression and in tension to determine the equilibrium and tensile moduli, respectively. Media stimulation resulted in a 2 to 5 fold increase in mechanical properties and a 2 to 3 fold increase in matrix accumulation in constructs over 6 weeks in culture. Benefits from mixing stimulation for collagen accumulation and compressive modulus appeared to peak near Re 2.9, and decreased with increased mixing intensity. This study suggests that fluid mixing can be optimized to enhance mechanical properties of anatomically-shaped engineered constructs.
Quantification of shape fidelity of complex geometries for tissue-engineered constructs has not been thoroughly investigated. The objective of this study was to quantitatively describe geometric fidelities of various approaches to the fabrication of anatomically shaped meniscal constructs. Ovine menisci (n ¼ 4) were imaged using magnetic resonance imaging (MRI) and microcomputed tomography (mCT). Acrylonitrile butadiene styrene plastic molds were designed from each imaging modality and three-dimensional printed on a Stratasys FDM 3000. Silastic impression molds were fabricated directly from ovine menisci. These molds were used to generate shaped constructs using 2% alginate with 2% CaSO 4 . Solid freeform fabrication was conducted on a custom openarchitecture three-dimensional printing platform. Printed samples were made using 2% alginate with 0.75% CaSO 4 . Hydrogel constructs were scanned via laser triangulation distance sensor. The point cloud images were analyzed to acquire computational measurements for key points of interest (e.g., height, width, and volume). Silastic molds were within AE10% error with respect to the native tissue for seven key measurements, mCT molds for six of seven, mCT prints for four of seven, MRI molds for five of seven, and MRI prints for four of seven. This work shows the ability to generate and quantify anatomically shaped meniscal constructs of high geometric fidelity and lends insight into the relative geometric fidelities of several tissue engineering techniques.
Replication of anatomic shape is a significant challenge in developing implants for regenerative medicine. This has lead to significant interest in using medical imaging techniques such as magnetic resonance imaging and computed tomography to design tissue engineered constructs. Implementation of medical imaging and computer aided design in combination with technologies for rapid prototyping of living implants enables the generation of highly reproducible constructs with spatial resolution up to 25 μm. In this paper, we review the medical imaging modalities available and a paradigm for choosing a particular imaging technique. We also present fabrication techniques and methodologies for producing cellular engineered constructs. Finally, we comment on future challenges involved with image guided tissue engineering and efforts to generate engineered constructs ready for implantation.
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